Accepted Manuscript

A compendium of wheat germ: Separation, stabilization and food applications

Fatma Boukid, Silvia Folloni, Roberto Ranieri, Elena Vittadini

PII: S0924-2244(18)30254-1
DOI: 10.1016/j.tifs.2018.06.001
Reference: TIFS 2242

To appear in: Trends in Food Science & Technology

Received Date: 14 April 2018

Revised Date: 1 June 2018
Accepted Date: 4 June 2018

Please cite this article as: Boukid, F., Folloni, S., Ranieri, R., Vittadini, E., A compendium of wheat
germ: Separation, stabilization and food applications, Trends in Food Science & Technology (2018), doi:
10.1016/j.tifs.2018.06.001.

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 ACCEPTED MANUSCRIPT
1 A Compendium of Wheat Germ: Separation, Stabilization and

2 Food Applications

3 Fatma Boukid 1,#, Silvia Folloni2, Roberto Ranieri2, Elena Vittadini1

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4 Food and Drug Department, University of Parma, Parco Area delle Scienze 27/A, 43124 Parma, Italy
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5 Open Fields S.r.L., Collecchio, Parma, Italy.

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6 # Corresponding author: Fatma Boukid, Food and Drug Department, University of Parma, Parco Area

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7 delle Scienze 27/A, 43124 Parma, Italy; e-mail: fatma.boukid@unipr.it; tel: +390521906079.

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18 Abstract

19 Background

20 Wheat germ is a precious by-product deriving from the milling industry, as it is a natural concentrated

21 source of essential amino and fatty acids, minerals, vitamins, tocopherols, and phytosterols. However,

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22 the presence of high enzymatic activities together with a high content of unsaturated oil, induce a fast

23 decrease in the nutritional value of wheat germ during storage and, consequently, strongly limit

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24 product’s shelf-life.

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25 Scope and approach

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26 In recent decades, flour blends from raw or/and processed wheat germ received great interest from
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27 nutritional and technological perspectives. Nevertheless, the quality of the end-product strongly

28 depended on the supplementation level, as well as the type and the severity of separation and
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29 stabilization techniques that wheat germ went through. Hence, in this review, the newest advances in

30 wheat germ pre-handling approaches and food applications are discussed to provide relevant and
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31 updated information about its worthiness to be a part of the human diet.

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32 Key findings and conclusions

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33 To fully valorize and preserve the nutritious potential of wheat germ, effective pre-treatments of

34 separation and stabilization are needed to guarantee its stability and suitability to meet food quality
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35 and safety standards. Such an underutilized ingredient might be a valuable fortifying component for a
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36 spectrum of foodstuffs.

37 Keywords: Wheat germ, Separation, Stabilization, Foodstuffs, Technological quality.

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40 State of the art of wheat germ

41 I.1. Wheat germ within the whole wheat

42 The whole wheat kernel is composed of three primary structures: endosperm, bran and germ.

43 Endosperm (80-85%) is mostly formed by starch and proteins and surrounded by several outer layers

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44 called “the aleurone”. The aleurone is made up of living cells enclosing bioactive compounds,

45 surrounded by thick cell walls composed of arabinoxylans and β-glucans, and some proteins (Y. M.

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46 Hemery et al., 2010).

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47 Bran (13-17%) is formed by pericarp and testa. The testa is a hydrophobic tissue rich in lignin and

48 characterized by the presence of lipidic compounds such as alkylresorcinols, present in a cuticle at the

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surface of this tissue (Y. M. Hemery et al., 2010). The pericarp lies in the outermost layers, which are
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50 composed of three layers, the epicarp, mesocarp, and endocarp (Yu et al., 2015).
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51 Wheat germ (WG) (2-3%) contains the embryo and the scutellum, which are separated from the

52 endosperm by the aleurone. The embryonic axis includes the radicle and plumule (Yu et al., 2015).
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53 The scutellum plays mainly the role of a storage organ. Also, it is the attachment organ between the
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54 pericarp and the embryo. Generally, most of the scutellum is discarded during wheat milling and the

55 “recovered germ” is mainly the embryo.

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56 I.2. Wheat germ: A precious milling co-product

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57 Together with the bran, WG is considered the most important by-product of the dry milling industry.

58 The world deposit of WG is estimated to be, annually, ca. 25,000,000 tons (Rizzello, Nionelli, Coda,

59 De Angelis, & Gobbetti, 2010), which is an important milling co-product to valorize. WG has been

60 shown to have important impact on human health (e.g. antioxidant, antihyperlipidemia,

61 hypocolesterolemic, anticancer effects) (Kumar et al., 2011; Mueller & Voigt, 2011; Ghafoor et al.,

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62 2017). WG may, indeed, be considered the most nutritious part of wheat grain because it contains

63 excellent amounts of nutrients such as proteins, unsaturated fatty acids, minerals, vitamins E and B,

64 dietary fiber, essential amino acids, flavonoids and sterols (Niu, Jiang, Pan, & Pang, 2013; Sun,

65 Zhang, Hu, Xing, & Zhuo, 2015; Zhu, Wang, & Guo, 2015) (See section III. Wheat germ chemical

66 composition). At present, WG is mainly used as a feed, while human consumption is still limited and

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67 not fully explored (Rizzello, Cassone, Coda, & Gobbetti, 2011). An emerging market is WG oil,

68 which is integrated in pharmaceuticals and cosmetic formulations or fertility agents (Gorusupudi &

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69 Baskaran, 2013; Niu et al., 2013). Defatted wheat germ (DWG) and wheat germ proteins are by-

70 products of the production of wheat germ oil (Niu et al., 2013). DWG contains about 30% proteins

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71 (Sun et al., 2015). Defatted wheat germ protein (DWGP) has a quality comparable to the model value

72 set by the FAO/WHO (Arshad, Anjum, & Zahoor, 2007) and to egg and milk proteins (Ge, Ni, Yan,

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Chen, & Cai, 2002). Hence, WG and its derivatives might be considered a valuable functional
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74 ingredient with high nutritional interest.
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75 I.3. Reasons for removing wheat germ during milling: Stability, Quality and Safety
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76 The widespread utilization of WG is limited by the rapid development of rancidity after isolation from
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77 the kernel (Li et al., 2016). The mechanical treatment involved in wheat milling favors oil diffusion,

78 and once the lipid fraction is exposed to the air, oxidation drastically increases (Geng, Harnly, &
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79 Chen, 2015; Gili, Palavecino, Cecilia Penci, Martinez, & Ribotta, 2017). The presence of unsaturated

80 fats as well as hydrolytic and oxidative enzymes favor WG rapid degradation (Xu et al., 2013).
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81 Enzymes, lipoxygenase and lipase, deteriorate lipids molecules generating acidity and volatile
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82 compounds (Capitani, Mateo, & Nolasco, 2011; Sjövall, Virtalaine, Lapveteläinen, & Kallio, 2000).

83 Therefore, once separated, WG shelf life is drastically shortened to a matter of few days, which is the

84 major limitation to WG industrial applications (Sjövall et al., 2000; Srivastava, Sudha, Baskaran, &

85 Leelavathi, 2006).

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86 Removal of intact WG in conventional milling process before kernel grinding is practically

87 impossible, especially in the case of soft wheat. WG can be gradually recovered after the first passage

88 through the rolling mills, but, being mainly crushed, WG is subject to oxidation and rancidity

89 phenomena. WG is, therefore, generally removed during milling of wheat kernels to increase the shelf

90 life of flour and to prevent occurrence of a strong flavor from rancid oils in the flour (Geng et al.,

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91 2015). Therefore, WG presence in flour negatively affects its shelf-stability and usability (Rizzello,

92 Nionelli, Coda, De Angelis, et al., 2010), and the final products stability and sensory attributes

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93 (Sjövall et al., 2000).

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94 Besides stability issues, WG contains some anti-nutritional components. Compared to refined flour,

95 unprocessed WG comes with higher anti-nutrients (i.e. phytic acid, agglutinins) concentrations. Phytic

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96 acid complexes minerals important for human nutrition thereby limiting their bioavailability (Demir &
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97 Elgün, 2014; Gupta, Gangoliya, & Singh, 2015; Zajdel, Wilczok, Węglarz, & Dzierżewicz, 2013).

98 Agglutinins, sugar binding proteins which increase intestinal permeability and might damage the gut
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99 lining, are also found in WG (de Punder & Pruimboom, 2013; Pellegrina et al., 2009). However, these

100 anti-nutrients can be drastically reduced by WG stabilization (de Punder & Pruimboom, 2013; Matucci
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101 et al., 2004).

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102 Being rich in fats, pesticide residues (e.g. organophosphorus insecticides, which are mainly used for

103 wheat conservation) tend to remain associated with WG because they are lipophilic (high affinity to
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104 fats) (Uygun, Koksel, & Atli, 2005; Uygun, Senoz, & Koksel, 2008). Pesticides and insecticides are
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105 serious threats to both human health and the environment (Hossard, Guichard, Pelosi, & Makowski,
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106 2017). Different approaches were developed to determine the quantities, and measuring the content of

107 these harmful chemical molecules, that often exceed the legal limits (Rezaei, Damalas, &

108 Abdollahzadeh, 2018). Therefore, food safety issues are another reason why the germ is removed and

109 not fully exploited. However, the final concentration of these residues depends on several factors such

110 as the cultivation area, the residual amount of pesticides in the soil, the duration of storage, and the

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111 processing (Uygun et al., 2005, 2008; Uygun, Senoz, Öztürk, & Koksel, 2009). The highest level of

112 pesticide residues legally tolerated in WG is 0.02 mg/kg for cyfluthrin (RD) and 0.05 mg/kg for

113 permethrin (European Food Safety Authority, 2016.).

114 In recent years, increasing efforts are being made to valorize milling “wastes” thereby turning the fate

115 of WG from waste to a highly valuable by-product. New strategies were developed to provide health-

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116 beneficial and low-cost ingredients for food-making. WG is a nutritious but unstable ingredient due to

117 the action of oxidative and hydrolytic enzymes on unsaturated fatty acids. Consequently, adequate

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118 techniques are needed to prolong WG shelf-life and preserve its nutrients. This comprehensive review

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119 is designed to acquire deep insights into the recent advances dealing with WG separation and

120 stabilization strategies, compositional traits, and food applications, through the use of scientifically-

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121 sound and reliable evidence.
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122 I. Advances in wheat germ separation
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123 WG separation might be performed throughout two main approaches: direct and gradual.
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124 Degermination is a direct approach, which might provide intact WG. As for the indirect approach, WG

125 is gradually separated in the form of flakes, through the successive passages throughout the milling
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126 process.
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127 II.1. Direct approach: Degermination

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128 The goal of the degerminator is to remove the germ without excessive grinding. Several types of
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129 degerminators have been patented (Table 1). Prior to degermination, a tempering phase of the grain is

130 required to reach a moisture level ranging from 19% to 25%, depending on the degree of

131 degermination. The degerminators first crush kernels from their thin edges to separate the germ and

132 endosperm without damaging the germ. Afterwards, endosperm undergoes successive passages of

133 breaking and reduction to obtain a fine flour. Such a pre-treatment has the advantage to reduce

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134 separation passages (purifiers to separate bran particles and germ from endosperm). In the mill

135 industry, until now, degerminators are not used in wheat processing, and they are mostly used in corn

136 processing to recover the germ (11% of the kernel) to be used for oil extraction (Gwirtz & Garcia-

137 Casal, 2014). Although WG represents only around 2% of wheat kernel, an important volume of WG

138 is annually removed worldwide. Indeed, wheat is ground throughout the world resulting in an

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139 important nutrient-dense by-product to be valorized. Therefore, more investigations are required to

140 optimize degermination technics (Rizzello, Nionelli, Coda, De Angelis, et al., 2010).

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141 II.2. Indirect approach: Gradual separation

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142 The objective of the milling process is to separate the bran and germ from the endosperm and to

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143 convert the endosperm into fine flour. WG is structurally a separate entity at the base of the kernel, but
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144 it is partly embedded in the endosperm. To properly separate WG from endosperm and avoid cell

145 walls break, the action of the machinery must be carefully adjusted during the milling phases, and WG
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146 separation is achieved gradually. As a matter of fact, milling diagrams have been evolved and

147 improved over the years, and therefore, their evolution is reported here following a chronological
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148 timeline.
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149 Primary processing of wheat: Single stone-grinding (Figure 1a)

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150 Stone-grinding is the traditional method used for wheat grinding. It consists of rubbing the grains

151 between two thick and heavy circular stones to produce whole wheat flour. Therefore, the
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152 conventional grinding process does not enable the separation between germ, bran, and endosperm,
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153 instead they are completely mixed together. The current milling diagrams of stone mills includes the

154 presence of a plansichter to separate the bran from the refined flour or at least to homogenize the size

155 of its particles.

156 Conventional method: Multiple grinding-sifting (Figure 1b)

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157 The milling process involves breaking, scraping and reducing grain to flour (Sakhare & Inamdar,

158 2014). Grain is progressively passed through grinders. During grinding, kernels are broken open and

159 the adhering endosperm is scraped off the bran layers. After each grinding passage, the middlings are

160 sifted and fractionated into fine endosperm, bran and germ, based on particle size. The remainings are

161 re-grinded and then re-sifted until the endosperm is milled into fine flour. In this case, a total

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162 separation of germ is not possible because some residual germ particles adhere to endosperm or bran.

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163 Modern processing of wheat: Break- Reduction System (Figure 1c)

164 Modern milling is based on 4 key-steps: 1) the break system; 2) the sizing system; 3) the reduction

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165 system, and 4) the tailings system (Sakhare & Inamdar, 2014).

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166 Tempering or conditioning is a step used to increase grain moisture content, which aids fractioning
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167 during the milling process (Hemery et al., 2010; Hemery, Rouau, Lullien-Pellerin, Barron, &

168 Abecassis, 2007). Cleaned kernels undergo a tempering phase to properly hydrate bran multi-layers
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169 cover which encases the germ and the endosperm (Hourston, Ignatz, Reith, Leubner-Metzger, &

170 Steinbrecher, 2017) and to favor detachment of the outer-layers of the kernel from the endosperm
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171 during the passages through the roller mills (Dennett & Trethowan, 2013).
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172 Break system is made of several successive grinding passages (a set of grinders/rollers). Rollers
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173 grinding consists of two series of roller. The first set of rollers operates at high speed to efficiently

174 grind the large particles of the endosperm. Then, for the streams rich in germ, the speed is reduced to
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175 prevent the deterioration of germ cells and the exposure of fatty acids. Therefore, the first grinding
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176 operation opens the tempered kernels to expose the endosperm and scrape a portion of the endosperm

177 from the bran (Sakhare & Inamdar, 2014). The second grinding allows the shearing off the germ in

178 flakes from the end of the grain, and the germ is collected as another fraction from the process

179 (Sakhare, Indrani, Inamdar, Gaikwad, & Rao, 2014).

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180 Sizing systems allow the separation of the large pieces of endosperm from smaller pieces of germ and

181 bran due to differences in the shape between the anatomic parts of the wheat kernel (Fistes & Rakić,

182 2015). Throughout the rollers, germ is pliable and tends to flatten, while bran forms low-density small

183 flakes (Posner & Hibbs 2005). As a result, the germ and bran are separated from each other, and

184 adhering endosperm is removed (Fistes, Tanovic, & Mastilovic, 2008).

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185 Reduction system is a set of rollers enabling the endosperm to be reduced in a finely white flour

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186 (Flamer 2008).

187 Tailing system is a set of purifiers, which ensure the separation of bran particles from the endosperm

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188 recovered from the previous systems to produce a flour free from germ and bran (Sakhare & Inamdar,

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189 2014). AN
190 Overall, the complete separation of the germ is difficult because there is no cleavage line between the

191 germ, aleurone layer and endosperm (Sakhare & Inamdar, 2014). The recovered germ is, actually, the
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192 embryonic axis of the wheat kernel, while scutellum is left attached to the bran (Finnie and Atwell

193 2016). The role of purifiers is important and requires constant monitoring and adjustment (Posner &
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194 Hibbs 2005).

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195 Modern processing of wheat: Debranning (Figure 1d)

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196 Nowadays, many industrial mills include the debranning process in durum wheat (T. turgidum subsp.
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197 Durum Desf.) milling because it increases mill capacity and enhances the quality of the semolina and
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198 milling yield (Bottega et al., 2009). Debranning is designed to remove a portion of bran ahead of the

199 first break, by friction and abrasion, without nicking the endosperm and limiting the number of broken

200 kernels (Bottega et al., 2009; Fistes & Rakić, 2014; Hemdane et al., 2015). A patented method of

201 milling process (US20060257541) with a debranning step started with wetting wheat kernels (to reach

202 a moisture content around 15%), intensely vibrated and then subjected to conditioning. Afterward, the

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203 conditioned kernels were decorticated by taking off the outer layers of bran. As a result, the kernel

204 contains less bran before milling, and consequently the separation of the germ after the first break unit

205 is facilitated. Therefore, such a preliminary step might enhance the recovery of WG in terms of quality

206 and quantity, but it might damage germ structure, if the grindings machines settings (i.e. distance

207 between cylinders and their rotation speed) are not optimized and some germ may be entirely removed

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208 during the debranning phase. Also, De Brier et al. (2015) showed that for common wheat (T. aestivum

209 L.) this process does not remove the kernel tissues homogeneously, and consequently a significant

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210 amount of starchy endosperm is lost. Thus, because of its more fragile/weaker structure, debranning is

211 generally not adopted for soft wheat milling. Therefore, the efficiency of germ separation depends on

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212 the shape and the form of the grain, the number of passages (grinding and sizing), and the know-how

213 of the miller.

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214 II. Wheat germ chemical composition
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215 Table 2 displayed the chemical composition of raw WG (RWG) taken from the public database of the

216 Danish Food Composition Databank (2009), which is considered among the most complete data bank
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217 of food nutrients. Beside genetic and environmental effects, compositional traits are closely associated
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218 with the processing used to isolate the germ and the degree of its extraction/recovery. Complete

219 information of WG composition taking into consideration all the environmental and processing factors
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220 affecting it are lacking. Nevertheless, WG is considered the most nutritious part of wheat kernel

221 providing 381 cal/100 g, with 54% provided by carbohydrates, 23% by proteins, and 23% by lipids
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222 (Nutrition data, 2014).

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223 Carbohydrates are the major component (51.3 g/100 g). A significant amount of dietary fiber (12.3

224 g/100 g) is also found, consistently with previous works (Marti et al., 2014; Sudha, Srivastava,

225 Leelavathi, & Leelavathi, 2007), which is characterized by a greater amount of insoluble (13.8–15.6

226 g/100 g) than soluble dietary fiber (2.8–3.6 g/100 g) (Srivastava et al., 2006). Important levels of

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227 proteins are reported (28.1 g/100 g), which is in the same range with previous works (Srivastava et al.,

228 2006; Sudha et al., 2007). Proteins consist primarily of albumins (34.5g/100 g), followed by globulins

229 (15.6 g/100 g), glutenins (10.6 g/100 g), and prolamines (4.6 g/100 g) (Zhu, Zhou, & Qian,

230 2006). These proteins have a well-balanced amino acid composition, with high levels of alanine,

231 arginine, asparagine, glycine, and lysine (Liu, Chen, Shao, Wang, & Zhan, 2017) , whereas cystine is

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232 lacking (Zhu et al., 2006). Further, this aminoacidic composition is close to the model value issued by

233 the FAO/WHO with good amino acid equilibrium (Arshad et al., 2007). Indeed, most of the essential

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234 amino acids are present in levels comparable to those found in egg and milk protein (Rizzello,

235 Nionelli, Coda, De Angelis, et al., 2010). Lipid content varied from 9.4 to 9.7 g/100 g (Table 2), which

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236 are slightly lower to those reported in Nutrition data (2014) (11.2-17 g/100 g), containing 1.4 g/100 g

237 saturated fatty acids, 1.1 g/100 g monounsaturated fatty acids and 4,3 g/100 g polyunsaturated fatty

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acids. Likewise, Rizzello et al. (2010) underlined that the lipids composition is rich in unsaturated
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239 fatty acids such as oleic, linoleic and α-linoleic acids suggesting WG oil can be marketed for direct

240 human consumption as dietary supplements (Giménez et al., 2013). Minerals are present at about 4.4
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241 g/100 g, which is almost four times more concentrated than in whole wheat kernel. An important
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242 amount of vitamins is also found. Notably, WG is considered the richest known source (plant origin)
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243 of vitamin E (Wang & Johnson, 2001), with α-tocopherol present in amount 16 times higher than in

244 any other fraction of wheat kernel (Gili, Palavecino, et al., 2017). WG contains also significant
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245 amounts of thiamine, riboflavin and niacin as well as phytosterols and polycosanols (Eisenmenger,

246 Nurhan, & Dunford, 2007; Engelsen & Hansen, 2009; Wang & Johnson, 2001). Moreover, it is a rich
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247 source of antioxidants mainly phenolics (Gelmez, Kıncal, & Yener, 2009; Zhokhov, Broberg, Kenne,
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248 & Jastrebova, 2010). The challenge, in isolating, storing and utilizing WG, is to maintain this high

249 nutritional quality by preventing lipid oxidation.

250 III. Wheat germ stabilization strategies

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251 Inactivation of oxidative enzymes (lipase and lipoxygenase) or/and removal of the oil fraction are

252 needed to ensure a stable WG (Sjövall et al., 2000). This can be achieved through implementation of

253 several processes that have been developed taking into consideration both producers’ and consumers’

254 request for easy, low-cost, effective, and safe processes. Several attempts were made to improve the

255 shelf-life of WG involving physical, chemical and biological approaches (Figure 2).

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256 IV.1. Physical stabilization

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257 WG physical stabilization might be subdivided into several sub-clusters: heat treatments (steaming,

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258 fluidization, spouted bed, and roasting/toasting) (Marti et al., 2014; Srivastava et al., 2006; Sudha et

259 al., 2007), radiation mediated treatment [microwaving (Xu et al., 2013), infrared radiation treatment

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260 (Gili, Palavecino, et al., 2017) and gamma radiation (Jha, Kudachikar, & Kumar, 2013; Li et al.,
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261 2016)] and thermal/mechanical treatments (cooking-extrusion and de-oiling) (Gómez, González, &

262 Oliete, 2012).

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263 Thermal stabilization

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264 The traditional methods of germ stabilization are based on thermal treatments (dry and wet) aiming at
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265 the inactivation of lipase thereby avoiding lipid oxidation. These enzymes are thermo-stable and

266 maintain more than 20% of their residual activity after heat treatment (60–90 °C, 1 h)
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267 (Kapranchikov, Zherebtsov, & Popova, 2004). Therefore, processing optimization is required to

268 completely inactivate lipase and to preserve the nutritional characteristics of WG.
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269 Steaming is one of the most efficient methods for WG stabilization, at it enables quick heating of the

270 product surface due to the high level of thermal energy transmitted per unit of time (Biglia, Comba,

271 Fabrizio, Gay, & Ricauda Aimonino, 2017). Steaming was shown to inactivate enzymes and enhance

272 WG shelf-life (Srivastava et al., 2006; Yan-bo et al., 2008). In particular, lipase activity was

273 completely inactivated, whereas the lipoxygenase was inactivated to an extent of 80–92%, after 15

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274 min at 125-130°C (Srivastava et al., 2006; Sudha et al., 2007). Quality indexes (such as fatty acid and

275 peroxide values) of WG during storage were found to be below the legal limits (e.g. fatty acid <

276 2 g/100 g of oleic acid; peroxide value < 15 meq O2/kg oil, Gili et al., 2017) after 30 days of storage

277 [i.e. 85 mg KOH/100g and 13.4 meq O2/kg after 30 days at 38°C, Xiaojun (2002)]. As for chemical

278 composition, steaming did not affect protein content, but it reduced fiber [RWG: 18.4%; steamed WG:

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279 15.8%] and vitamin E [RWG: 15.8 mg / 100 g; steamed WG: 10.71 mg / 100 g] (Srivastava et al.,

280 2006).

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281 WG stabilization efficiency was increased by the combination of steaming with other dry heating

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282 techniques (e.g. hot air drying, baking oven, and microwave). Lab-scale results revealed that lipase

283 was inactivated in all cases, but lipoxygenase activity was reduced by different degrees depending on

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284 the combined techniques used (Sudha et al., 2007). Residual lipoxygenase activity of steamed WG
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285 (14.9%) was slightly higher than steaming-coupled to oven drying (100 °C for 2–3 h) (14.2%), and

286 microwave oven at 750 W for 4–5 min (12.98%) (Sudha et al., 2007). Consistently, atmospheric steam
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287 (5 min) combined with microwave (320 W, 3 min) resulted in a 3.7% residual lipoxygenase activity

288 (Su-fang et al., 2008). Ying (2008) optimized treatment conditions as follows: steam treat for 5 min,
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289 560 W microwaves treat for 5 min after cooling. Steamed germ was dried in baking oven at 200 °C for
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290 8–12 min resulting in the lowest residual of lipoxygenase (7.9%) (Sudha et al., 2007). Hence,

291 combinations efficiently extended the storage life of WG (Ying 2008). However, they negatively
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292 affected the vitamin E content that was reduced from 15.8% (RWG), to 13.6% in steamed and baked

293 oven dried, to 12.2% in steamed and hot air oven dried, and to 11,9% in steamed and microwave dried
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294 wheat germs (Srivastava et al., 2006).

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295 Fluidization is an operation to keep solid particles moving in an upward direction in a flow of fluid

296 (gas or liquid) (van Ommen, Valverde, & Pfeffer, 2012). Fluidization, provides intense heat and mass

297 transfer between material and the hot air, which implies a uniform treatment in the fluid-bed (Gili,

298 Torrez Irigoyen, Penci, Giner, & Ribotta, 2017; Srivastava et al., 2006). Lab-scale fluidized bed

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299 (240°C, 1 min) decreased moisture content in WG from 11.4% to 2%. Such a treatment allowed

300 complete lipase inactivation, whereas lipoxygenase residual activity was 13.55%±1.3 (Sudha et al.,

301 2007). Furthermore, WG showed a lipoxygenase residual activity of 16.00%±1.4 after 60 days of

302 storage at room temperature (Srivastava et al., 2006). A multistage fluidized bed with 45°C as outlet

303 temperature resulted in low level of oxidation (peroxide value = 7.2 mmol/kg and acid value =18.47

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304 mg/g) after 30 days of accelerated storage (temperature of 40°C, relative humidity of 85%) (Bin et al.,

305 2010). However, vitamin E content was reduced of about -35% (Srivastava et al., 2006). Overall,

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306 although this treatment proved high efficiency in WG stability, huge investments are necessary for

307 processing facilities, which limits its industrial application (Xu et al., 2013)

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308 Spouted bed is a special type of fluidized bed, where the fluidizing gas is injected from a central spout.

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309 Using a laboratory-spouted bed drier, WG was treated for 180–540 s with air at 140–200 °C. Results
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310 showed a drastic reduction of lipase (65%) and lipoxygenase activities (91%) (Yöndem-Makascioǧlu,

311 Gürün, Dik, & Kincal, 2005). Peroxide values were found less than 10 meq O2/kg after storage (20
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312 weeks at room temperature). Processed-WG (200 °C for 360 s) showed golden color and nutty flavor

313 and acceptable sensory evaluation (Yöndem-Makascioǧlu et al., 2005). Moreover, spouted bed has a
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314 uniform air circulation, and consequently such a treatment is suitable for WG flakes stabilization due
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315 to their non-sticky surface and low density (Gili, Torrez Irigoyen, et al., 2017).
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316 Roasting is commonly used for stabilizing WG and whole grain flour because it reduces moisture

317 content and the activity of enzymes involved in lipid peroxidation (Ramadan, Showky, & Sulieman,
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318 2008). Roasting develops a particular flavor and golden color, which are associated with the Maillard
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319 reactions (Ciarmiello et al., 2013). Several WG roasting methods have been patented (US 3783164 A,

320 US 3036919 A, US 20070292583 A1, US 8173193 B2…). Ramadan et al. (2008) investigated the

321 influence of lab-scale roasting (in a drying oven at 160 °C for 20 min) on the WG lipid fraction.

322 Results revealed that total lipid recovery and fatty acid composition were not influenced, and

323 oxidation phenomena was not detected. Furthermore, ethanol extracts of roasted WG (at 160 °C for 20

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324 min) had strong total polyphenols content and antioxidant activities including free radical scavenging

325 against DPPH and ABTS radicals, FRAP, and ORAC of WG (Krings, Johansson, Zorn, & Berger,

326 2006; Zou, Yang, Zhang, He, & Yang, 2015). Gelmez et al. (2009) extracted antioxidants from roasted

327 WG using supercritical carbon dioxide extraction (pressure: 336 bar, temperature: 58 °C and duration

328 10 min) (Gelmez et al., 2009) and recovered 6 mg GAE phenolics/g extract, 57.3 mg scavenged DPPH

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329 /g extract, and 6.7 mg tocopherol/g extract (almost complete recovery). Such high extraction levels

330 were possible thanks to partial cell wall destruction and consequent release of some bound phenolic

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331 compounds, or/and formation of Maillard reaction products (Zou et al., 2015).

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332 Microwave heating

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333 Chun-feng et al., (2006) results showed that microwave radiating could inactivate wheat germ lipase

334 and prolong its storage stability. The increase of processing time decreased water content in WG and
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335 increased protein and fat contents (Xiao-hong et al., 2007). Youssef et al. (2010) found also an
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336 important increase in vitamin A (57%), vitamin E (40%) and vitamin C (8%). The fatty acids

337 composition of oil was not affected, and no trans fatty acids were developed (Xiao-hong et al., 2007).
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338 Previous works (Rodríguez-López et al., 1999; Yan-bo et al., 2008; Xu et al., 2013) highlighted the
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339 efficiency of microwave in WG stabilization because, compared to traditional heating methods, it

340 combines both thermal and radiation effects. Yan-bo et al., (2008) revealed that microwaved WG did
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341 not show any significant change in the odor or flavor after 7 weeks of storage (Sjövall et al., 2000).

342 Besides, it had lower residual lipase activity than dried WG (Yan-bo et al., 2008). In conventional
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343 heating, WG absorbs energy generated by the thermal gradients through convection, conduction and
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344 radiation (Gutiérrez, Catalá-Civera, Bows, & Peñaranda-Foix, 2017). However, microwave radiation

345 directly interacts with WG, and not with the surrounding medium, reducing energy costs (Ciarmiello

346 et al., 2013; Gutiérrez et al., 2017)

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347 To date, no reports on industrial application of WG stabilization using microwave are available.

348 Recently, a self-designed continuous microwave processing equipment was used to investigate WG

349 stabilization process (Xu et al., 2013). It was reported that the line process (capacity: 20 kg/h,

350 microwave power: 4 Kw, processing time: 8 min, and ventilation rate: 60 N m3/h) allowed a complete

351 deactivation of lipoxygenase and a reduction of 89% of lipase activity (Xu et al., 2013). Also, low

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352 oxidation during storage was reported (acid value increase of 6.56% after 60 days) (Xu et al., 2013).

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353 Infrared radiation

354 Infrared radiation intensity of 4800 W/m2 for 3 min was the best processing condition to stabilize WG

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355 because its tocopherol content was not reduced, and its shelf life was extended to at least 90 days

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356 (25°C in a temperature-controlled room) with respect to RWG (15 days) (Gili, Palavecino, et al.,

357 2017). Consistently, the individual and total tocopherols contents were insignificantly (p > 0.05)
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358 affected by the radiation at 70–90°C. WG treated at 90°C for 20 min retained more than 96.43% of its
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359 original tocopherols (Li et al., 2016). Moreover, irradiated WG at 90°C for 20 min acquired the

360 optimal stabilization effect (fatty acids content and peroxide value remaining below 5% and 2.24 meq
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361 O2/kg for 60 days at 40°C), and its residual lipase and lipoxygenase activity were 18.02% and 19.21%,
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362 respectively (Li et al., 2016). The residual water activity was also very low (0.13 %), and about

363 96.42% of WG original tocopherols was preserved (Li et al., 2016). Likewise, positive effects were
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364 found on total polyphenol content and antioxidant activity (Bilgiçli, Demir, & Yılmaz, 2014). Overall,

365 compared with conventional heating, infrared treatments had important advantages including uniform
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366 heating, short heating time, low quality losses and low energy consumption (Gili, Palavecino, et al.,
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367 2017). It is also a rapid and effective tool for retarding free fatty acids development (Li et al., 2016).

368 Gamma radiation (γ-irradiation)

369 Few studies were carried out on WG stabilization by means of γ-irradiation. γ-irradiation treatment

370 enabled stabilization of WG through a 31.2 % inactivation of lipase (Jha et al., 2013), and it (0–30
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371 kGy doses) did not induce significant changes in the chemical composition (moisture, total ash, crude

372 fat, free fatty acid, protein) of WG. However, an increase in free fatty acids content was found in

373 irradiated WG samples (Ramadan et al., 2008). The impact of γ-irradiation (at radiation doses 1/4, 1/2

374 and 1 kGy) and roasting at (160 °C for 20 min) on lipids was compared (Ramadan et al., 2008), and it

375 was found that roasting and irradiation treatments did not significantly affect the total lipid recovery

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376 nor the fatty acid composition of WG lipids (Ramadan et al., 2008). Lipids from irradiated WG

377 exhibited relatively stronger radical scavenging potential than lipids from roasted WG (Ramadan et

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378 al., 2008). Also, γ-irradiation could be applied for WG preservation using a relatively low dose (1/4

379 kGy), and it was shown to have an effect in reducing the antigenicity of agglutinins (Ramadan et al.,

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380 2008), also confirmed in vivo mice studies (Vaz et al., 2012).

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381 Thermo-mechanical stabilization
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382 Cooking-extrusion is a multi-step, multi-functional thermal/mechanical process used in a large number
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383 of food applications (Kaur, Sharma, Singh, & Dar, 2015). In the case of WG, a cooking-extrusion

384 treatment significantly improved its stability. Indeed, the content of free fatty acids of WG after
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385 storage (12 days) remained within the safe level (i.e. lower than 1.06 g/kg) (Lin, Huff, & Hsieh, 2002).
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386 According to patent method (US20090155439), extrusion comprises the following steps: the separated

387 germ is firstly loaded into a mechanical extruder applying the suitable settings for enzyme
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388 denaturation; maintaining a residence time suitable for stabilization; and water is then added to

389 maintain a level of moisture content. Free fatty acid levels were assessed to check the efficiency of the
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390 procedure. When free fatty acid content is 5% or lower, germ may be considered stable. The
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391 maximum temperature reached by the WG mass in the final section of the extruder, was 180 °C

392 (Gómez et al., 2012). Besides, a reduction of the microbial content and of some anti-nutritional

393 components, and partial inactivation of enzymes were observed (Gómez et al., 2012).

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394 De-oiling or de-lipidation or defatting is a thermal/mechanical process aiming at the removal of the

395 lipid fraction of WG. According to the patent of Grandel (1959), WG de-oiling was carried out by

396 inserting fresh WG in a hydraulic press and compressing it until it contains less than 4% oil. Defatted

397 WG may then be ground using a roll mill to extract the remaining oil. The obtained defatted germ was

398 subjected to an alkaline treatment using a solution of sodium carbonate and finally the germ slurry was

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399 flaked on a roller dryer at 130-140°C. In this case, de-oiling might lead to degradation of heat-

400 sensitive compounds due to high temperatures during processing (Panfili, Cinquanta, Fratianni, &

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401 Cubadda, 2003). It was also reported that fresh WG was steamed at 60-70° and then oil was

402 mechanically extracted (up to 50%) using a double stage compression screw press (Eng, Mubarak, El-

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403 Nono, Owies, & Saad, 2012) .

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404 Refrigeration technology
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405 Refrigeration technology might be used as a tool to preserve and reutilize WG (Capitani, Mateo, &
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406 Nolasco, 2011; Sjövall, Virtalaine, Lapveteläinen, & Kallio, 2000). The recuperated WG from the

407 milling process was carefully cleaned and immediately cold stored to preserve freshness and avoid
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408 rancidity. The refrigerated product was later added to semolina during pasta-making. The whole
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409 process could occur within 1 week. Such a technology is rarely used at an industrial scale probably

410 because of the expenses of refrigeration equipment’s.

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411 VI.2. Chemical stabilization

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412 Enzyme denaturation

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413 Lipase and lipoxygenase are the enzymes of concern in WG stability. Lipases have optimal activity at

414 ∼pH 7.0–8.0 which decreases on either side of this pH range (Ahmad, Fatima, Khan, & Khan, 2010).

415 As for lipoxygenase, optimal activity is reached at ∼pH 4.0-10.0 and decreases outside this range

416 (Sudharshan, Srinivasulu, & Appu Rao, 2000). Enzyme inactivation can, therefore, be achieved by

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417 either acidification (hydrochloric acid, acetic acid, calcium hydroxide, Sharma et al., 2014) or

418 alkalization [0% decrease in lipase activity at pH=10 (Rao, Rajendran, Rajeshwara, & Prakash, 1991)

419 or 93% at pH=12 (Rajendran, Rao, & Prakash, 1990)]. Enzymes inactivation using chemicals enabled

420 an oil recovery of 90% as compared to mechanical de-oiling (50%) (Singh & Ric 1979). However,

421 using chemical extraction is not always approved for food formulations because traces of toxic

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422 solvents might be detected in the oil.

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423 Oil removal

424 WG oil removal can be achieved by either organic solvent (e.g. hexane) or supercritical CO2

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425 extraction.

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426 Compared to organic solvents, supercritical CO2 (SC-CO2) extraction had similar yields [(92% oil
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427 recovery after 3 h of processing at pressure of 38 MPa, temperature 55 °C, particle size of 0.35 mm

428 and a CO2 flow of 1.5 dm3·min−1, (Panfili et al., 2003)); (80% oil recovery after 8 hours of lab-
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429 processing at pressure of 3 MPa, temperature 40 °C, around 8 h, (Piras et al., 2009)); (79% oil

430 recovery after 90 min using a lab-scale extraction (pressure 26.2 MPa, temperature 40-50°C, CO2 flow
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431 2 mL min-1), (Ge, Sun, Ni, & Cai, 2000))]. Also, oils extracted using supercritical CO2 are solvent-
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432 free and do not require the traditional refining processes. Regarding chemical composition, defatting,

433 using food-grade solvent such as hexane, increased WG protein content (up to 38%), soluble fiber
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434 (from 2.07 to 3.01%), and insoluble fiber (from 14.4 to 24.49%) (Bansal & Sudha, 2011), consistently

with Ma et al., (2014). As well, oils extracted using supercritical CO2 preserve a high-quality nutrients
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435

436 composition (free fatty acids, 12.4%; tocopherol content, 416.7 mg tocopherol/g wheat germ oil).
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437 Finally, CO2 is nontoxic, nonflammable, noncorrosive, cheap, and recyclable (Gòmez & de la Ossa,

438 2000). Such technology is very competitive to the conventional chemical extraction, because it

439 simplifies the oil refinement and eliminates the solvent distillation stage (Gòmez & de la Ossa, 2000).

440 However, using a continuous cycle on an industrial scale, great attention should be paid to the particle

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441 size of WG because it greatly influences the yield (Panfili et al., 2003). Indeed, the reduced size of

442 WG particles might induce their agglomeration resulting in the reduction of oil extraction (Panfili et

443 al., 2003).

444 IV.3. Biological stabilization

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445 Recently, fermentation of WG by lactic acid bacteria (e.g. sourdough fermentation) was proposed as a

446 mean for WG stabilization through enzyme inactivation as consequence of acidification (Marti et al.,

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447 2014; Rizzello, Nionelli, Coda, Di Cagno, & Gobbetti, 2010) and the process was compared to

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448 toasting (Marti et al., 2014). Acidification induced by LAB was found to inactivate lipase and

449 lipoxygenase, as does the toasting process (Marti et al., 2014; Rizzello, Nionelli, Coda, Di Cagno, et

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450 al., 2010) thereby increasing WG shelf-life. Furthermore, volatile compounds deriving from lipid
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451 oxidation (determined using SPME/GC/MS analysis) were reduced in LAB fermented WG, as

452 compared to non-fermented WG, after a 40-day storage (Rizzello, Nionelli, Coda, De Angelis, et al.,
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453 2010). Total phenols of RWG (0.49 ± 0.02 mM gallic acid equivalent) was lower than that of

454 fermented WG (0.65 ± 0.02 mM gallic acid equivalent) (Rizzello et al., 2011). Fermentation also
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455 increased total antioxidant activity [from 6.83 to 8.50 (mmol Trolox equiv/g)], total free amino acids
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456 [from 1.36 to 1.68 (mg/kg)] and in vitro protein digestibility [from 62.92 to 65.11%] (Rizzello et al.,

457 2010). Regarding antinutrients, LAB fermentation decreased raffinose (Rizzello, Nionelli, Coda, De
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458 Angelis, et al., 2010) and glutathione (Marti et al., 2014) presence as compared to unprocessed WG.

459 As a result, dough enriched with fermented WG enhanced its rheological quality in terms of stability
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460 during mixing and development (Marti et al., 2014).

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461 Table 3 displays a summary of the advantages and dis-advantages of the above discussed approaches

462 for WG stabilization.

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463 IV. Technological implication of wheat germ inclusion in wheat flour

464 blends

465 As previously highlighted, (see section III. Chemical composition of wheat germ), several bioactive

466 components (e.g. tocopherol, minerals and amino acids) of nutritional interest are located in the germ.

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467 Germ removal, therefore, induces the loss of most of these compounds, making refined wheat flour

468 and its derived cereal products, nutritionally-poor in micronutrients and dietary fiber. In recent years,

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469 more cereal-based foods have been enriched with WG or its derivatives to obtain products with

470 upgraded nutritional value (Ma et al., 2014; Zhu, Lian, Guo, Peng, & Zhou, 2011).

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471 V.1. Bread

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472 Bread is a staple food throughout the world, and the increasing trend toward healthy eating contributed

473 in increasing interest in including bran and germ in many food items and, bread in particular. Indeed,
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474 several studies demonstrated that the use of WG enhanced the nutritional value of bread, by increasing

475 the concentration of minerals, proteins, fat and dietary fiber as compared to bread prepared from
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476 refined flour (De Vasconcelos et al., 2013; Rizzello et al., 2013, 2011; Rizzello, Nionelli, Coda, De
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477 Angelis, et al., 2010; Rizzello, Nionelli, Coda, Di Cagno, et al., 2010; Sidhu, Al-hooti, & Al-saqer,

478 1999). Fortification greatly affected dough and bread quality depending on the level or form (raw or
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479 processed) of the WG used (M. Majzoobi, Farhoodi, Farahnaky, & Taghipour, 2012; Shin, Kim, &

480 Kim, 2013).

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481 Wheat germ supplementation impact on dough rheological properties

482 The effect of WG fortification on the bread-making performance of flour may be evaluated based on

483 dough rheological properties that provide valuable information on dough workability and expected

484 product quality.

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485 RWG is usually avoided in bread formulation because it was reported to have a negative impact on the

486 viscoelastic properties of dough (Ma et al., 2014). RWG incorporation at 10% level, notably decreased

487 dough tenacity and stability (M. Majzoobi, Farhoodi, et al., 2012), consistently with the finding of

488 Gòmez et al. (2012). Ma et al. (2014) reported that RWG supplementation (1-11%) increased the wet

489 gluten content of dough (from 33 to 34.5%) as compared to 100% wheat flour (32.5%). However,

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490 Srivastava et al. (2007) found that RWG addition (5, 10, 15 and 20%) had no significant effect on

491 water absorption of the dough. Using a farinograph, Majzoobi et al. (2012) found that the dough made

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492 with RWG had low water absorption, low consistency, and short stability. Gómez et al. (2012)

493 reported that RWG integration weakened dough due to the presence of glutathione, which is able to

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494 break disulphide bonds in the gluten and to decrease the stability and strength of the dough (Al-Hooti,

495 Sidhu, Al-Saqer, & Al-Othman, 2002). Such result might be attributed to the enzymes and reducing

496
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agents in RWG, which might contribute to the alteration of the network of proteins, starch and water,
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497 reducing dough viscoelasticity (Ma et al., 2014).
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498 Several methods have been proposed, to inhibit glutathione activity in RWG, including heating,

499 fermentation and antioxidant addition, which showed different degrees of success (Majzoobi et al.,
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500 2012). Heat treatment (150°C for 45 minutes) increased water absorption, consistency, and stability of
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501 WG enriched dough compared to RWG (Gómez et al., 2012). Also, blends of wheat flour with

502 different amounts of heat-treated WG (70°C, 2h) increased water absorption, from 64.5 (3%) to 65.7%
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503 (12%) compared to 100% wheat flour-based dough (Sun et al., 2015). Toasted WG was found to

504 reduce dough development time as well as stability beyond a 5% addition (Srivastava et al., 2006).
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505 The high lipid content in WG might decrease stability having a lubricating and softening effect on the
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506 dough (Srivastava et al., 2006). A decrease in the falling number was reported after the

507 supplementation of different levels of heat-treated WG (3%, 6%, 9% and 12%) indicating high α-

508 amylase activity in flour causing high starch degradation, and subsequent pasting temperature

509 increase, and viscosity decrease compared to the control (Sun et al., 2015). This might be due to the

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510 activation of the residual α-amylase or/and the extensive gelatinization during the heat-treatment

511 (Gómez et al., 2012; Srivastava et al., 2006; Sudha et al., 2007). Addition of extruded WG (2.5, 5, 7.5,

512 10, and 20 %, flour basis) did not affect rheological properties (increased development time and

513 decreased stability after over-kneading, dough tenacity, extensibility, and dough alveographic

514 strength) (Gómez et al., 2012; Majzoobi et al., 2012). Lactic acid bacteria fermentation greatly

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515 contributed to decrease glutathione content in WG with dough exhibiting high stability during mixing

516 and development up to substitution of flour with fermented WG at 20 g/100 g level (Marti et al.,

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517 2014). A 4% substitution of sourdough fermented WG did not modify the water absorption capacity

518 and had higher softening value (42 ± 3 and 38 ± 2 BU) compared to the conventional dough (35 ± 3

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519 Brabender Units, BU) (Rizzello, Nionelli, Coda, De Angelis, et al., 2010; Rizzello, Nionelli, Coda, Di

520 Cagno, et al., 2010).

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521 Summarizing, the weakening of dough strength in flour blends-containing different raw/treated WG

522 might be strongly related to the dilution of gluten content (Sun et al., 2015). RWG was less
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523 appreciated than processed WG due the high amount glutathione involved in dough weakening (Zhu et

524 al., 2006). Besides the influence of addition level, rheological properties were impacted by the
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525 previous treatment of WG. Compared to physical treatments, lactic acid bacteria fermentation
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526 enhanced the dough properties. In all cases, beyond 20% WG addition, dough quality is severely

527 damaged (Gómez, Oliete, Rosell, Pando, & Fernández, 2008; Srivastava et al., 2006).
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528 Wheat germ supplementation impact on bread quality

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529 The effect of WG integration on bread-making is strongly related to bread type (leavened, steamed,
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530 sourdough or flat), the WG pre-treatment, and, obviously, the level of incorporation.

531 Regarding leavened bread, RWG addition increased bread crumb firmness (> 305%) with respect to

532 white flour bread (210.1±12.0g) (Al-Hooti et al., 2002) as well as it decreased specific loaf volume

533 (Sidhu et al., 2000). However, using a mix of additives (30 ppm potassium bromate and 50 ppm
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534 ascorbic acid), specific loaf volume was improved at levels of 10% of RWG. Another mix of additives

535 (0.5% sodium stearoyl-2-lactylate, 30 ppm potassium bromate and 50 ppm ascorbic acid) enabled to

536 obtain a 20% RWG-enriched bread, which was appreciated by consumers (Houti et al., 2002).

537 As for processed germ, volume even increased (Gòmez et al., 2012) and firmness decreased (Li et al.,

538 2016) in the presence of small amounts (up to 5%) of extruded WG, suggesting that extrusion might

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539 be a suitable treatment to stabilize WG for bread applications. However, at higher levels, WG

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540 decreased bread loaf volume (Gòmez et al., 2012). Up to 10 % WG, loaf color was comparable to the

541 control bread, which might be probably attributed to the carotenoids from WG (Al-Hooti et al., 2002;

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542 Gómez et al., 2012). Brownness also increased with WG supplementation due to protein and free

543 sugar content increase accelerating the Maillard reaction. Gòmez et al. (2012) reported that 10% raw

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544 or extruded WG showed similar sensory scores. More pronounced differences were more evident at
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545 higher level of supplementation.
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546 Considering sourdough bread, the addition of 4% sourdough fermented WG flour decreased crumb

547 firmness , resilience and fracturability, without diminishing bread acceptability (Marti et al., 2014;
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548 Rizzello, Nionelli, Coda, Di Cagno, et al., 2010). Furthermore, they underlined a special flavor and
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549 taste attributed to bread made with 4% RWG and 4% fermented WG. A salty attribute also

550 characterized its sensory profile, probably due to lactic acid bacteria acidification and proteolysis
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551 (Rizzello, Nionelli, Coda, Di Cagno, et al., 2010). Fermented WG was reported to extend shelf life by

552 the synthesis of antifungal compounds, resulting in reduced lipase activity and, consequently lower
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553 lipid oxidation (Marti et al., 2014; Rizzello, Nionelli, Coda, De Angelis, et al., 2010; Rizzello,
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554 Nionelli, Coda, Di Cagno, et al., 2010).

555 Steamed bread supplemented with 1-3% RWG or DWG (chemically defatted using hexane) had

556 similar elasticity, cohesiveness and resilience, but relatively higher crumb firmness and chewiness to

557 flour-based bread (Ma et al., 2014). Higher supplementation (11%) resulted in small specific volume,

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558 high crumb firmness and low elasticity due to the decrease in gas holding capacity during dough

559 fermentation (Ma et al., 2014). Supplementation might have probably interfered with optimal gluten

560 matrix formation during fermentation and steaming (Sun et al., 2015). Compared to RWG, DWG had

561 higher negative effect on crumb firmness, probably, due to its higher protein content responsible of

562 high water absorption. On the other hand, RWG is characterized by high amylase activity, which

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563 increased the availability of reducing sugars in the dough, thereby promoting fermentation. Bread

564 with 1-3% RWG had higher a* and b* values and was consequently yellow and red color were more

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565 intense than breads with DWG. Low supplementation enhanced bread color, indicating that an

566 appropriate amount of WG can improve the color of steamed bread (Ma et al., 2014). Such result

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567 might be attributed to Millard reactions products or/and the presence of WG antioxidants (e.g. alpha-

568 tocopherols) reducing the effect of oxidative enzymes (lipoxygenase and polyphenoloxidase) in the

569
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flour (Bahal, Sudha, & Ramasarma, 2013). Furthermore, sensory acceptability was improved with the
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570 application of low level of WG flour (3%) (Ma et al., 2014), while over 9% taste and texture were

571 hampered (Sun et al., 2015).

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572 As for flat bread, increasing amounts of heat-treated (at 150°C for 45 min) WG (0-20%) increased
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573 crumb firmness, and decreased volume (Majzoobi et al., 2012). Compared to RWG, processed WG
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574 had a lower impact on texture because of the reduction of glutathione (around 50% by heat treatment)

575 and better gluten preservation (Majzoobi et al., 2012). According to sensory analysis, taste, crust color
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576 (light golden brown) and flavor were enhanced (up to 15% addition) due to the Maillard reactions

577 products resulting from germ sugars and proteins interaction (Majzoobi et al., 2012). The color of
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578 bread surface was reported to become darker with increased addition level (up to 30%) (Levent,
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579 Bilgiçli, & Ertaş, 2015). According to sensory evaluation, beyond 10% level incorporation hindered

580 the overall acceptability scores of flat breads.

581 Regardless of bread type, low addition (<5%) of germ (both raw or processed) did not affect bread

582 quality nor diminish the acceptability of the bread, yet it enhanced its nutritional quality (Majzoobi et

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583 al., 2012; Marti et al., 2014). In terms of texture, RWG increased bread crumb firmness, while in terms

584 of flavor and taste, it conferred an appreciated sweet note to the bread. Beyond 20% WG substitution,

585 undesirable effects were observed resulting in low acceptability. Processed wheat germ showed

586 similar effects on bread, consisting in mainly increasing firmness. Yet, sourdough fermented WG

587 bread showed some particular advantages, which is correlated with decreasing firmness. Concluding,

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588 besides WG nutritional added value, low levels (<5%) enhanced color and taste, with no effect on

589 texture. If improvers are also incorporated in bread formula, around 10% or 20% WG can be the limit

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590 of addition to levitated bread (Gòmez et al., 2012) and up to 10% to those unleavened (Al-Hooti et al.,

591 2002; Sidhu et al., 1999).

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592 V.2. Pasta and noodles

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593 The global pasta and noodles consumption is upward worldwide, and, consequently, improving their

594 nutritional quality is important for consumers seeking health-beneficial food (Huh et al., 2017;
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595 Monteiro et al., 2016; Vijayakumar & Boopathy, 2014). Pasta and noodles may look alike, but they

596 have many differences: (i) the origin: Pasta is European and noodles are Asian; (ii) raw ingredients:
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597 pasta is primarily made from durum wheat semolina, while noodles are made with common wheat
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598 flour; and (iii) processing: pasta is extruded or laminated, while noodles are made using the "roll-and-

599 cut" method (Lu et al., 2005; Fu 2008; Monteiro et al., 2016).
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600 To enhance their nutritional composition or/and to give new taste and flavor, several ingredients (e.g.
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601 legumes, vegetables, eggs, and spices) are occasionally incorporated in pasta and noodles formulation.
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602 Currently, the use of wheat bran and germ in pasta fortification is receiving great interest due their

603 interesting nutritional quality (Aktaş, Bilgiçli, & Levent, 2015; Pınarlı, İbanoğlu, & Öner, 2004; Tarzi,

604 Shakeri, & Ghavami, 2012). Regardless of the fortification, pasta must maintain a desirable firm

605 texture during cooking (Kaur, Sharma, Nagi, & Dar, 2012). WG enhanced the nutritional quality (ash,

606 protein, fat and mineral contents) of pasta (Aktaş et al., 2015; Pınarlı et al., 2004; Tarzi et al., 2012).

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607 Raw and microwaved WG supplemented-macaroni had similar in vitro protein digestibility compared

608 to the conventional product (Pınarlı et al., 2004). As for noodles, 15% DWG flour enhanced

609 nutritional properties, in terms of amino acid composition, minerals, and B vitamins (Ge, Sun, Ni, &

610 Cai, 2001).

611 Wheat germ supplementation impact on pasta/noodles quality and stability

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612 WG incorporation in pasta (10-20% WG) and noodles (15% defatted WG) resulted in higher red and

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613 lower yellow indexes compared to the control sample (Aktaş et al., 2015; Tarzi et al., 2012), probably

614 because of occurrence of non-enzymatic browning related to Maillard or carbonyl-amino, reactions

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615 developed between proteins and sugars (Zhang, Ames, Smith, Baynes, & Metz, 2009)

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616 WG supplementation increased cooking loss of macaroni (15%: from 7.8 to 11.6 %, Pınarlı et al.,
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617 2004) and noodles (30%: from 5.8 % to 7.3 %, Aktaş et al., 2015), probably related to the disruption

618 of the gluten matrix (Aktaş et al., 2015; G. Kaur et al., 2012; Pınarlı et al., 2004; Tarzi et al., 2012).
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619 Because of the high cooking losses beyond 20%WG addition, 15% was indicated as the maximum

620 level of acceptable enrichment (Pınarlı et al., 2004).

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621 Sensory evaluation of cooked noodles found the highest mouthfeel scores at 10 % WG addition (Aktaş

622 et al., 2015). Also, 10-20% raw and microwaved supplemented macaroni had similar texture, taste and
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623 overall score as compared to the control (Pınarlı et al., 2004; Tarzi et al., 2012). Beyond 20% WG,

624 sensory scores decreased (Aktaş et al., 2015). Furthermore, the use of 15 % DWG in noodle
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625 formulation enhanced the chewing properties as well as the smooth appearance (Ge, Sun, Ni, & Cai,
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626 2001).

627 Microbiologically, macaroni is generally considered a safe product due to its low water activity.

628 Macaroni sensory attributes made from semolina blended with 15% raw and microwaved WG did not

629 change significantly after a year of storage (Pınarlı et al., 2004). However, after production, Tarzi et al.

27
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630 (2012) showed that pasta enriched with 10-20% RWG had higher amounts of bacteria (275±50

631 (cfu/g)) than pasta made 100% wheat flour (control sample) (20±00 (cfu/g)) and pasta made with heat

632 treated WG (110 °C for 35 min) (2.5±5 (cfu/g)). Until 6 months of storage, no significant difference

633 was observed between pasta made with 15% heat treated WG (110 °C for 35 min) and control sample

634 in terms of aerobic bacteria, yeast and molds. Yet after 12 months, increasing molds and yeasts were

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635 found in the heat treated WG supplemented pasta (100±81.650 (cfu/g) compared to the control

636 (50±50.73 (cfu/g)) (Tarzi et al., 2012). This result suggested that pasta enriched with heat treated WG

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637 can be safely stored until 6 months.

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638 V.3. Baked goods

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639 Cookies fortified with 5-15% WG had increased protein, fat, mineral, and fiber contents (Petrović et
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640 al., 2017), cakes had high mineral contents (Ca, Cu, Fe, Mn, P, K and Zn) (Levent & Bilgiçli, 2013),

641 and muffins showed important increase in Mg (Thomason, 1999). Biscuits fortified with 20% WG had
M

642 high crude fiber as well as vitamins C, Folic acid, A, and E (Youssef, 2015). Likewise, DWG is a

643 natural high-grade protein and amino acid fortifying agent (Arshad et al., 2007; M. Majzoobi,
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644 Darabzadeh, & Farahnaky, 2012), and 25% fortification increased protein (around 30%) rich in
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645 essential amino acids especially lysine content (around 2 g/100 g) (Arshad et al., 2007).

646 Wheat germ supplementation impact on dough rheological properties

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647 WG addition increased the water required to make the dough of biscuits or cookies due to protein and
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648 fiber present in WG (Bansal & Sudha, 2011; Petrović et al., 2015). Such an increase might result in the
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649 formation of strong structure with high extensibility and hardness (Bansal & Sudha, 2011; Majzoobi,

650 Darabzadeh, et al., 2012). Up to 15% WG incorporation had no significant influence on the

651 rheological performance of the cookies’ dough (Petrović et al., 2015). In addition, WG particle size

652 greatly affected the rheological characteristics (Petrović et al., 2015). Majzoobi, Darabzadeh, et al.

653 (2012) studied the impact of WG particle size on dough quality and found that increasing the particle
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654 size of WG from 280 to 1.195 µm) caused an increase in batter consistency and density, but still

655 retaining workability. Indeed, the addition of WG (up to 20%) at small particle sizes (280 µm) resulted

656 in the most appreciated product by consumers due to (Majzoobi, Darabzadeh, et al., 2012).

657 Wheat germ supplementation impact on baked goods quality

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658 Crust and crumb color of cakes were significantly affected by WG addition (Levent & Bilgiçli, 2013),

659 with cake crust becoming darker (low luminance and high red index) with increasing levels of WG.

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660 Such result might be attributed to the acceleration of sugar caramelization and Maillard reactions

661 (Arshad et al., 2007). On the other hand, Majzoobi et al. (2012) found that, while the addition of WG

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662 from 0 to 20% enhanced crumb yellowness (b*), crust color was unaffected. On the other hand,

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663 Majzoobi et al. (2012) found that, while the addition of WG from 0 to 20% enhanced crumb

664 yellowness, crust color was unaffected. Consistently, Petrović et al. (2017) observed that increasing
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665 WG level (5-15 %) increased yellowness but color was more dependent of particle size (150-2000 µm)
M

666 than amount of germ. They found 15% as the highest level of germ ADDITION and particle size of

667 150 µm as the most suitable conditions for production of cookies with the highest yellowness (b*).
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668 WG incorporation (lower than 15%) caused a decrease in cake’s volume but did not significantly
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669 change its textural properties (Majzoobi, Darabzadeh, et al., 2012). Integration of WG beyond 15%

670 enhanced softness and springiness of the cakes but did not affect cohesiveness (M. Majzoobi,
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671 Darabzadeh, et al., 2012; Petrović et al., 2017). Furthermore, increasing WG flour particle size

increased cake’s volume and uniformity index (Levent & Bilgi, 2013), and batter density (M.
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672

673 Majzoobi, Darabzadeh, et al., 2012). It was also reported to decrease cookies’ hardness (Petrović et
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674 al., 2017),

675 Sensory analysis of cakes showed that 0-10% of WG addition did not significantly influence overall

676 acceptability of the product, while at higher levels, taste and texture quality attributes were

677 significantly decreased (M. Majzoobi, Darabzadeh, et al., 2012). Regarding cookies, up to 15%
29
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678 substitution of wheat flour with DWG produced acceptable products similar to the control (100%

679 wheat flour) (Arshad et al., 2007). However, beyond 15% DWG, overall acceptability was low, due to

680 crumbly texture and beany flavor (Arshad et al., 2007). Cookies made from 20-30% WG were highly

681 acceptable (Al-Marazeeq & Angor, 2017; Levent & Bilgiçli, 2013). Beyond 40% level, enrichment

682 resulted in predominant flavor of WG (Bansal & Sudha, 2011).

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683 V.4. Beverages

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684 Fermented products are gaining popularity due the increasing trend towards functional foods. WG

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685 based-fermented products might be considered a promising new functional beverage to expand the

686 market (Mueller & Voigt, 2011), as proven by the presence of several supplements fortified with WG

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687 with prebiotic effects currently available on the market (Boros, Nichalatti, & Shoenfeld, 2005; Coda et

al., 2011; Comı́n-Anduix et al., 2002; Judson et al., 2012; Mueller, Jordan, & Voigt, 2011; Mueller &
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688

689 Voigt, 2011; Otto et al., 2016).

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690 Abbas et al. (2015) prepared two dairy by-products, buffalo’s butter milk and sweet whey that
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691 included freshly milled WG. The products, supplemented with 2% WG had higher yellowness,

692 apparent viscosity and antioxidant activity, but lower lightness compared with non-supplemented
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693 control. Results indicated that 2% WG was the most accepted fortification level as for the sensory

694 properties (Abbas, Husse, Seleet, Bayoumi, & El-Azi, 2015). Furthermore, Seleet et al. (2016)
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695 developed a novel yoghurt-like fermented product fortified with WG (0-40%). Indeed, the addition of
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696 WG slurry enhanced acid development during fermentation and increased the viscosity of the
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697 fermented products. They also suggested that 20% addition of WG enabled the production of

698 fermented milk-based product without any adverse effect on its quality (Seleet, Assem, Abd El-

699 Gawad, Dabiza, & Abd El-Salam, 2016). However, further investigations are required to explore their

700 stability during storage and consumer acceptability.

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701 Tarhana is a wheat-yoghurt fermented mixture, widely consumed in turkey (Kumral, 2015). Bilgiçli et

702 al. (2007) investigated Tarhana enrichment with up to 50% WG to improve its nutritional status:

703 increasing WG level resulted in increased content of protein, minerals, and total phenolic compounds.

704 Phytic acid content also increased after WG addition, but it was significantly reduced (almost 90%) by

705 fermentation (Bilgiçli & İbanoğlu, 2007). On the other hand, higher amounts of WG incorporation (up

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706 to 50%) resulted in darker color (Bilgiçli & İbanoğlu, 2007). Therefore, 10% WG was the preferred

707 substitution level judged by consumers (Nermin Bilgiçli, İbanogˇlu, & Herken, 2007)

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708 Different levels (0-10%) of WG were incorporated also in a fresh chilled dairy dessert causing an

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709 increase in dry matter and a decrease in pH (Majzoobi, Ghiasi, & Farahnaky, 2016). WG enhanced

710 darkness, hardness, cohesiveness and gumminess, while springiness decreased. Sensory tests

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711 demonstrated that WG (10%) resulted in low acceptability, which might be related to the desserts’
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712 wheaty taste, unpleasant smell and hard texture. Therefore, chilled dairy dessert-containing 5 and

713 7.5% WG received the highest mouthfeel scores due to the improved texture.
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714 V.5. Other emerging wheat germ applications

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715 Coffee substitution

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716 Several odorants of roasted WG are present in the aroma of roasted coffee such as: 2-methylbutanal,
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717 3-methylbutanal, 2-methylfuran and 2,3- butanedione in high concentration (El‐Saharty, El‐Zeany,

718 Tawakkol, & Berger, 1998; Fadel, Abdel Mageed, & Lotfy, 2008). A mixture of milled chicory roots
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719 (Cichorium intybus L.) and wheat germ (1:1 w=w) was roasted (0.5 h, 180 °C), extruded and then
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720 milled. The obtained mixture was subject to a first sensory analysis (similarity of sensory attributes

721 with a real coffee), which identified more than 90 volatile components in both real coffee and coffee

722 substitute. A second sensory test (odor profile analysis) revealed that the substituted coffee had similar

723 aroma and odor as compared to coffees. Odor profile also showed that the sweetish=caramel-like note

724 scored higher in substituted-coffees than in those prepared with 100% roasted chicory. During storage,
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725 the enrichment with WG supplemented-coffee decreased Strecker aldehydes and diketones

726 concentrations, while it increased phenolic compounds (Fadel, Abdel Mageed, & Lotfy, 2008).

727 Busbousa

728 Al Shehry (2015) studied WG (0, 5, 10 or 15%) addition to Busbousa, an Arabic local dessert

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729 consisting of semolina, sugar, milk and butter. The results showed WG supplemented-Busbousa had

730 higher amounts of protein, fat, fiber and ash, and lower antinutritional components than those made

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731 from semolina. Also, sensory evaluation revealed a supplementation level of 10% was the most

732 appreciated (Al Shehry, 2015).

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733 V. Concluding remarks and future trends

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734 Over the last years, a trend toward WG consumption is increasing due to its interesting nutritional

735 traits such as α-tocopherol, vitamin B, dietary fiber, polyunsaturated fats and minerals. However, WG
M

736 is often considered unfit for human consumption for stability and safety issues.
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737 A major obstacle in WG utilization is its low stability, and consequently effective methods were
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738 developed to prevent its rapid deterioration and to prolong its shelf-life for further processing. Also,

739 the presence of antinutrients limited the widespread of WG. Therefore, removing WG from the kernel
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740 went through an evolutionary process to improve the separation as well as to provide higher yield and

741 better nutrients composition. However, several difficulties are encountered during the separation of
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742 WG limiting its usability. Therefore, innovative methods and equipment’s are needed to overcome
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743 industrialization limits. As for stabilizing approaches, several methods were developed, with different

744 grade of efficiency. Physical methods were reported to be the most efficient in enzymes deactivation,

745 but they might deteriorate the nutritional value due the high temperature or prolonged treatment.

746 Mechanical methods were considered not fully resolving and required the integration of heat

747 treatment. However, chemical methods are still suspicious, especially, from consumers demand for

32
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748 natural, solvent-free, safe and minimally-processed products. Hence, biological methods are suitable

749 for consumers’ requirements, but more work is needed to ameliorate the effectiveness of bioprocesses

750 in reducing lipid adverse reaction. WG and its derivatives are considered attractive functional and

751 flavoring ingredients for foodstuffs making. Yet, their overall acceptability is strongly related to WG

752 status (raw or processed), the level of addition, and the quality requirements of the supplemented-food.

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753 Notably, WG oil is widely appreciated for its pharmaceutical and nutritional value, but it is still

754 underutilized in the food field.

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755 WG safety is another serious critical point to be carefully considered, since WG might contain levels

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756 of contaminants (e.g. pesticides and insecticides), exceeding the standard limits limiting its usability.

757 Yet, given the mounting trends towards biological agriculture, important amount of organic wheat is

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758 now available. Therefore, organic WG might be a safer raw ingredient for food fortification.
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759 Conflicts of interest:
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760 The authors declare no conflict of interest.

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761 Author Contributions:

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762 Fatma Boukid collected the data and compiled and wrote the manuscript. Silvia Folloni, Roberto

763 Ranieri and Elena Vittadini planned, drafted and corrected the manuscript.
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764 Abbreviations:
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765 WG: wheat germ

766 RWG: raw wheat germ

767 DWG: defatted wheat germ

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768 DWGP: defatted wheat germ protein

769 References

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1162 Maillard reaction and the analysis of protein glycation by mass spectrometry: probing the
1163 pathogenesis of chronic disease. Journal of Proteome Research, 8(2), 754–769.
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1164 https://doi.org/10.1021/pr800858h
1165 Zhokhov, S. S., Broberg, A., Kenne, L., & Jastrebova, J. (2010). Content of antioxidant hydroquinones
1166 substituted by b-1,6-linked oligosaccharides in wheat milled fractions, flours and breads.
https://doi.org/10.1016/j.foodchem.2009.12.084
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1167
1168 Zhu, K.-X., Lian, C.-X., Guo, X.-N., Peng, W., & Zhou, H.-M. (2011). Antioxidant activities and total
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1169 phenolic contents of various extracts from defatted wheat germ. Food Chemistry, 126(3), 1122–
1170 1126. https://doi.org/10.1016/j.foodchem.2010.11.144
1171 Zhu, K.-X., Wang, X.-P., & Guo, X.-N. (2015). Isolation and characterization of zinc-chelating
1172 peptides from wheat germ protein hydrolysates. Journal of Functional Foods, 12, 23–32.
1173 https://doi.org/10.1016/j.jff.2014.10.030
1174 Zhu, K.-X., Zhou, H.-M., & Qian, H.-F. (2006). Proteins Extracted from Defatted Wheat Germ:
1175 Nutritional and Structural Properties. Cereal Chemistry Journal, 83(1), 69–75.
1176 https://doi.org/10.1094/CC-83-0069
1177 Zou, Y., Gao, Y., He, H., & Yang, T. (2018). Effect of roasting on physico-chemical properties,
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1178 antioxidant capacity, and oxidative stability of wheat germ oil. LWT, 90, 246–253.
1179 https://doi.org/10.1016/j.lwt.2017.12.038
1180 Zou, Y., Yang, M., Zhang, G., He, H., & Yang, T. (2015). Antioxidant Activities and Phenolic
1181 Compositions of Wheat Germ as Affected by the Roasting Process. Journal of the American Oil
1182 Chemists’ Society, 92(9), 1303–1312. https://doi.org/10.1007/s11746-015-2689-1
1183
1184 Figure caption:
1185 Figure 1: Wheat germ separation as a function of milling diagram evolution: The case of durum wheat.

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1186 This figure underlines the evolution of milling process from a simple to more sophisticated one to
1187 allow a better separation of endosperm, brand and germ
1188 Figure 2: Wheat germ stabilization approaches. This figure summarizes the main methods to stabilize
1189 wheat germ.

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1214 Figure 1:Wheat germ separation as a function of milling diagram evolution: The case of durum wheat.
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WG stabilization

Physical Chemical Biological

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Thermal Alkalis Fermentation

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Radiation Acids

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Thermal/ Supercritical carbon
mechanical dioxide
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1217 Figure 2: Wheat germ stabilization approaches.

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1218 Table 1: Degerminators patents.

Patent Publication date Applicant Title

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Method of degerminating a kernel of grain by simultaneously compressing the edges of
US4189503 19 Feb 1980 Cereal Enterprises, Inc.

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the kernel

Method and apparatus for degerminating a grain kernel by impelling the kernels along a

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US4301183 17 Nov 1981 Cereal Enterprises, Inc.
guide vane into an impact surface

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US4365546 28 Dec 1982 Cereal Enterprisess, Inc. Apparatus for degerminating a kernel by compressing the edges of the kernel

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US5250313 5 Oct 1993 Cereal Enterprises, Inc. Grain milling and degermination process

WO2003047366A1 12 Jun 2003 Satake Usa, Inc. Corn degermination process

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US6936294 30 Aug 2005 Satake Usa, Inc. Corn degermination process

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US6953165 11 Oct 2005 The Quaker Oats Company Corn milling process

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US7553507 30 Jun 2009 Satake Usa, Inc. Corn debranning and degermination process
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EP3124119A1 1 Feb 2017 Cereal Enterprises, Inc. Degerminator

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1222 Table 2: Basic chemical composition of WG.

Content (/100 g) Unit Content Range

Energy Cal 381
Protein, total g 28.1 27.7 - 28.4
Fat, total g 9.6 9.4 - 9.7
saturated fatty acids g 1.4

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monounsaturated fatty acids g 1.1
polyunsaturated fatty acids g 4.3
Carbohydrate, total g 51.3 51.3 - 51.3

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Carbohydrate, available g 39
Starch g 26.1

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dietary fiber g 12.3 10.4 - 14.1
Moisture g 6.7 6.6 - 6.8

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Vitamins
Vitamin E α-te 11.0
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alpha-tocopherol mg 11.0
Vitamin K µg 39
Vitamin B1, thiamine Mg 1.45
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Vitamin B2, riboflavin mg 0.61

Vitamin B6 mg 1.42 1.36 - 1.47
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Minerals g 4.4 4.3 - 4.5

Sodium, Na mg 5.5 4.90 - 6.00
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Potassium, K mg 1046 1000 – 1120

Calcium, Ca mg 41 37.0 - 47.0
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Magnesium, Mg mg 259 236 – 276

Phosphorus, P mg 1000 960 – 1100
Iron, Fe mg 5 4.10 - 5.30
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Copper, Cu mg 1.1 1.00 - 1.10

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Zinc, Zn mg 17.8 14.4 - 27.0

Iodine, I µg 0.25 0.150 - 0.400
Manganese, Mn mg 18.0 11.0 - 20.0
Chromium, Cr µg 1.0 1.00 - 2.00
Selenium, Se µg 3 1.00 - 13.0

1223 Danish Food Composition Databank. http://www.foodcomp.dk/v7/fcdb_default.asp

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1224 Table 3: Stabilization methods of wheat germ: advantages and dis-advantages.

Advantages Dis-advantages

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Physical
Steaming Reduction in oxidation phenomena (Biglia et al., 2017); complete lipase Incomplete lipoxygenase inactivation (Biglia and others 2017); high
activity inactivation (Sudha, Srivastava, Leelavathi, & Leelavathi, 2007), residual water content in the product (Sudha and others 2007;

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prolonging WG self-life (Xiaojun, 2002; Yan-bo et al., 2008); quick heat up of uninform convective heating (Biglia et al., 2017); high costs for
product surface due to the high quantity of thermal energy transmitted per unit processing facilities (Sudha et al., 2007).
of time (Biglia et al., 2017); possibility of operation with air at higher

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temperatures (Sudha et al., 2007).
Fluidization, Complete lipase inactivation (Sudha et al., 2007); no fatty acid composition Important investment for processing facilities (Gili, Torrez Irigoyen,
spouted bed modification (Bin et al., 2010); possibility of simultaneous roasting (Sudha et Penci, Giner, & Ribotta, 2018; Sudha et al., 2007).

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al., 2007); good air circulation (Gili, Torrez Irigoyen, Penci, Giner, & Ribotta,
2017).

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Roasting/Toasting Reduction of moisture content and lipid peroxidation (Demir and others 2003). Reduction of phenolics content with longer roasting time;
development of a particular flavor and golden color (Ciarmiello and others development of Maillard reaction products (Zou et al., 2015).
2013); no significant modification of lipids (Ramadan 2008); increased total

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polyphenols content and antioxidant activities (Zou, Gao, He, & Yang, 2018);
WG storability extension (Krings and others 2000).
Microwave Lab-scale: no nutrient and low-quality losses, uniform heating, product Industrial application: low radiation penetration and non-uniform

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sterilization, short processing time and energy saving (Xu et al., 2013); low heating (Campañone & Zaritzky, 2005).
environmental impact (Campañone & Zaritzky, 2005).

Infrared radiation
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Low quality losses, uniform heating, short processing time, and low energy
consumption (Gili, Palavecino, Cecilia Penci, Martinez, & Ribotta, 2017); low
Incomplete lipase and lipoxygenase inactivation, increase in free
fatty acids and peroxide value of WG oil (Li et al., 2016).
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WG water activity; retarding lipid oxidation (Ciarmiello et al., 2013; Li et al.,
2016); micronutrients preservation (e.g. tocopherols total polyphenol content)
(Demir & Elgün, 2014; Li et al., 2016)
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Gamma radiation Shelf-life stability (Ramadan et al., 2008; Jha et al., 2013); no change chemical Incomplete lipase inactivation (Jha et al., 2013).
composition; stronger radical scavenging potential than lipids from roasted
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WG (Ramadan et al., 2008); reduction in allergic reactions towards WG

agglutins (Vaz et al., 2012).
Extrusion cooking Reduction of the microbial content, reduction some anti-nutritional factors Modification of chemical composition, nutritional (reduction in
(Gómez, González, & Oliete, 2012); improved the stability of WG during vitamin content...) and functional properties (Gómez et al., 2012).
storage (Lin, Huff, & Hsieh, 2002); A valid alternative to heating treatments
(Gómez et al., 2012).
De-oiling Preservation of nutritional composition (protein content, soluble fiber, and Low yield, only up to 50% oil recovery (Eng et al., 2012);

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insoluble fiber) (Bansal & Sudha, 2011); solvent-free and stable process (Eng, degradation of heat-sensitive compounds (Panfili et al., 2003).
Mubarak, El-Nono, Owies, & Saad, 2012).
Chemical
Alkalis/acids High oil recovery (90%) (Singh & Ric, 1979). Traces of toxic chemical; incomplete lipase deactivation (Ahmad et
al., 2010).

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Supercritical High yield (Panfili et al., 2003); preservation of nutritional composition High costs; WG granule size must be controlled to void
carbon dioxide (Panfili et al., 2003; Gómez & de la Ossa, 2000); solvent-free oil (Gómez & de agglomeration (Panfili et al., 2003).
la Ossa, 2000); reduction in peroxidase and lipoxygenase activities simple oil

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refinement (there is no need to a distillation stage) (Gómez & de la Ossa,
2000).

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Biological
Fermentation Solvent-free, enzyme deactivation; increased shelf-life; decrease lipid Incomplete lipase and lipoxygenase inactivation (Marti et al., 2014);
oxidation; decrease in some antinutrients, (raffinose and glutathione) (Marti et

U
al., 2014; Rizzello, Nionelli, Coda, De Angelis, & Gobbetti, 2010).

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1225

1226 Figure 1:Wheat germ separation as a function of milling diagram evolution: The case of durum wheat.

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WG stabilization

Physical Chemical Biological

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Thermal Alkalis Fermentation

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Radiation Acids

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Highlights

• Wheat germ is valuable by product of the milling industry.

• Wheat germ separation is a crucial step for its valorization.

• Suitable processing may preserve and prolong wheat germ quality.

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• Process optimization can provide a functional ingredient.

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