International Journal of Food Microbiology 112 (2006) 75 – 80 www.elsevier.


Development of a new oat-based probiotic drink
Angel Angelov a,⁎, Velitchka Gotcheva a , Radoslava Kuncheva a , Tsonka Hristozova b

Department of Biotechnology, University of Food Technologies, 26 Maritza Blvd, 4002 Plovdiv, Bulgaria b Institute of Microbiology, Bulgarian Academy of Sciences, 26 Maritza Blvd, 4002 Plovdiv, Bulgaria Received 31 May 2005; received in revised form 8 February 2006; accepted 27 May 2006

Abstract In the present work, a whole-grain oat substrate was fermented with lactic acid bacteria to obtain a drink, combining the health benefits of a probiotic culture with the oat prebiotic beta-glucan. The levels of several factors, such as starter culture concentration, oat flour and sucrose content, affecting the fermentation process, were established for completing a controlled fermentation for 8 h. The viable cell counts reached at the end of the process were about 7.5 × 1010 cfu ml− 1. It was found that the addition of sweeteners aspartame, sodium cyclamate, saccharine and Huxol (12% cyclamate and 1.2% saccharine) had no effect on the dynamics of the fermentation process and on the viability of the starter culture during product storage. Beta-glucan content in the drink (0.31–0.36%) remained unchanged both throughout fermentation and storage of the drink. The shelf life of the oat drink was estimated to 21 days under refrigerated storage. © 2006 Elsevier B.V. All rights reserved.
Keywords: Oat fermentation; Oat drink; Probiotic; Lactic acid bacteria; Beta-glucan

1. Introduction Continuous development of new functional foods is the response of science and industry to the increased consumer awareness regarding health and the role of foods for improving quality of life (Bland and Medcalf, 1994; Blades, 2000; Verschuren, 2002). Long known for its benefits, oats is becoming popular as part of a healthy diet and new oat products emerge at the functional food market. Most probiotic foods at the markets worldwide are milkbased and very few attempts are made for development of probiotic foods using other fermentation substrates such as cereals. Their large distribution and important nutritive value have focused the attention on their use as raw materials for the development of new fermented functional foods. Oats and barley are major sources of beta-glucan, recognized as the main functional component of cereal fibers. Studies have indicated the hypocholesterolaemic effect of this compound, leading to 20–30% reduction of LDL-cholesterol, and to an expected overall effect of reduced cardiovascular disease risk (Stark and
⁎ Corresponding author. Fax: +359 32 644 102. E-mail address: (A. Angelov). 0168-1605/$ - see front matter © 2006 Elsevier B.V. All rights reserved. doi:10.1016/j.ijfoodmicro.2006.05.015

Madar, 1994; Wrick, 1994; Gallaher, 2000). The cereals with highest beta-glucan content are oats and barley (Wood and Beer, 1998; Manthey et al., 1999). In 1997, the FDA of US has officially acknowledged as functional the products made of whole-grain oats or oat fiber with a minimum of 0.75 g betaglucan/serving size. The low glycaemic index of oat products is especially important for diabetics, and the ingestion of betaglucan-containing viscous foods is reported to affect the level of fat emulgation in the gastro-intestinal tract and reduces lipase activity (Anderson and Bridges, 1993; Wrick, 1993, 1994; Stark and Madar, 1994). The release of low-molecular fatty acids during beta-glucan fermentation in the colon preconditions its potential anti-carcinogenic effect (Oku, 1994; Salminen et al., 1998; Gallaher, 2000). In addition, beta-glucan is known as prebiotic, stimulating the growth of some beneficial residential colon microorganisms such as bifidobacteria (Jaskari et al., 1998; Wood and Beer, 1998). Marklinder and Lonner (1992), as well as Johansson et al. (1998) reported that after appropriate processing, oats is a suitable substrate for fermentation with lactic acid bacteria. Enzymatically treated oat bases have been developed and applied by Màrtensson et al. (2001, 2002) as substrates for lactic acid fermentation with dairy starter cultures.

9–4. Germany) at 4–6 °C. Test was performed also with adding combinations of sucrose and sweeteners as follows: A) 1.04% (w/v) sodium cyclamate (E952) + 0. 2. All sweeteners are products of Esarom. The different fermentation rates could be attributed both to strain specificities and differences of oat media. Viable cell enumeration Enumeration of viable cells of L. 3. Austria. plantarum B28 aiming to achieve the required levels of viable cells in probiotic products — at least 106–107 cfu ml− 1 (Mattila-Sandholm et al. Statistical analysis Each experiment was performed in four separate trials.0 and 10% inoculum concentrations applied. Oat substrate The oat mash used as a substrate was prepared from wholegrain oat flour “Mina Prim” (oat cultivar “Mina”. The cell count of the culture was 1011 cfu ml− 1.0. beta-glucan content and the viability of the starter culture. 1999). 5% and 10% (v/v) starter culture.0 and 10% starter culture suspension of L.008% (w/v) aspartame (E951). 1. 2.b.1. 4 °C). Darmstadt. which gave inoculation levels of 9. The strain was maintained on MRS-agar (E.m. plantarum B28 was performed by estimation of colony forming unit number on MRS-agar plates (medium pH 5. Co. Starter culture concentration The oat mash was inoculated with 1. / International Journal of Food Microbiology 112 (2006) 75–80 The aim of the present work was to develop a synbiotic functional drink from oats by combining a probiotic starter culture and whole-grain oat substrate.7. Angelov et al.8 × 108 and 7.5 and 7.0. Storage observations were carried out at 4–6 °C for 24 days.0% or 10% inoculum resulted in reaching the desired pH levels of 4.5 after much longer fermentation (16 h) of an oat base with commercial mixed dairy cultures. As these levels were above the required for probiotic products.0% (w/v).2% saccharine. Values of TA around 1. 2. 3. B) 1. 2. respectively. 2. Waltham. These results show that in terms of acid formation 8 h is an appropriate fermentation time. Ireland). Short fermentation time was preferable in order to minimize the risk of contamination. Marklinder and Lonner (1992) achieved pH 4.5 in significantly shorter time — 8 and 4 h.H. USA) was used. Food grade sucrose in concentrations of 1. the probiotic strain L. France) at λ = 510 nm.0% inoculum. The appropriate concentrations of oat flour.0–4. Starter culture was obtained by overnight incubation at 37 °C in MRS-broth. Bulgaria) and tap water in different concentrations 4. Results and discussion Based on previous research. while Martensson et al.6 × 107. as well as sweetener effect on fermentation and storage. Refrigerated storage was carried out for 24 days with periodical observations of pH. respectively. 5.5% sucrose + 0. Determination of beta-glucan Beta-glucan content was evaluated by an enzymatic kit according to the manufacturer's instructions (Megazyme International Ireland Ltd. Data was submitted to one-way analysis of variance (ANOVA) with a least significant difference of 95%.0% (w/v) was added to the slurry. Fermentation and storage The oat mash was inoculated with 1%. 1. The culture was centrifuged (4500 g. Results represent the means with standard deviations. plantarum B28 was selected for fermentation of the heat treated oat mash.. compared to 1. . 2002). 2000.. Starter culture A probiotic strain — Lactobacillus plantarum B28 was used in the study.005% (w/v) saccharine (E954) + 0.3 × 109 and 7. titratable acidity and dry matter For the measurement of pH ORION 420A pH-meter (Thermo Electron Corp.5 for 6 h when fermenting oatmeal soups with L. respectively.5% sucrose + 0. Results from the fermentations are presented in Fig.4. and titratable acidity (TA) was determined by titrating 10-ml samples with 0.1 N NaOH.3–4. Materials and methods 2. 2. phenolphthalein used as indicator. The strain was isolated from a cereal-based fermented drink and selected as probiotic in previous studies (Gotcheva et al. Ales. 2. Merck. plantarum strains.8. With 5.7) after incubation at 37 °C for 48 h. The slurry was then heated at 95 °C for 10 min and cooled to 37 °C.2. Fermentation was carried out at 37 °C for 6–10 h. 5. Wicklow.. Blagopoluchie... Huxol contains 12% cyclamate and 1.5.0 and the number of viable cell counts was above 106 cfu ml− 1. Estimation of shelf life The shelf life of the fermented oat drink was defined as the period of refrigerated storage (4–6 °C) during which pH remained above 4.0. respectively. starter culture and sucrose had to be estimated. it was more reasonable to use 5.3. Germany) and C) 1.5% sucrose (control). Absorbance was measured by spectrophotometer Anthelie Light (Secomam. and the shelf life of the drink. washed in distilled water and re-suspended in distilled water to its original volume.76 A. 2. 2.0% starter culture concentration.0% inoculum. 1993). Essenzanfabrik G. TA.6.1 × 108 cfu ml− 1. MA.0 and 5.5% (v/v) Huxol (Veelman. in 6 h the levels of viable cell counts reached 9. 10 min. TA was expresses as °N (degrees Neuman) (Vangelov and Karadjov.1.5°N were reached for 6 and 8 h with 10.5 × 1010 cfu ml− 1. Determination of pH.5 and 2. (2002) reported pH 3. The application of 5.

Results from testing three oat flour concentra- tions (4. With the lower flour concentrations applied. Changes of pH (A). Other authors reported pH 4. Changes of pH (A). Effect of the oat flour concentration on oat mash fermentation (average inoculum level — 2 × 108 cfu ml− 1). Effect of starter culture concentration on oat mash fermentation. viable cell counts (B) and betaglucan content (C).5 and 7. / International Journal of Food Microbiology 112 (2006) 75–80 77 Fig.5 after 16 h fermentation Fig. Angelov et al.5 were registered in about 6 h of fermentation. Acid production was lowest when oat flour consisted of 7. 2. pH values below 4. 1. Concentration of oat flour Concentration of whole-grain oat flour in the mash is important for the fermentation and the beta-glucan content in the oat products.0. 2. 5. 3.0–4.A. titratable acidity (B) and viable cell counts (C). .0%) are presented in Fig.52 in 8 h.0% mash and pH values reached 4.2.

The increase of flour content influenced the viable cell counts as well.5 and 15 h to Fig. . When 4.0–5. Variant C: sucrose + aspartame + sodium cyclamate + saccharine. / International Journal of Food Microbiology 112 (2006) 75–80 Fig. Effect of sweeteners on oat mash fermentation (average inoculum level — 2 × 108 cfu ml− 1).41 log orders. These results show that 4.0 and 5.. 3.65 and 2. respectively. Changes of pH (A).0–4. Variant B: sucrose + Huxol. viable cell counts (C). Angelov et al. 2000) probably due to the higher dry matter content of the medium — 16–18%. titratable acidity (B) and viable cell counts (C). Effect of sucrose concentration on oat mash fermentation (average inoculum level — 2 × 108 cfu ml− 1). Marklinder and Lonner (1992) fermented 5% w/w oatmeal soups for 6 h to reach pH 4. Changes of pH (A). the increase of cell counts was 2. of different oat substrates (Màrtensson et al. titratable acidity (B).5% oat flour content in the mash is more appropriate for intensive fermentation. Variant A: sucrose.78 A. 4.5% oat flour were used.

5% were not significant.5 was reached on the 6th hour. oat flour — 5. / International Journal of Food Microbiology 112 (2006) 75–80 79 Fig. The aim was to achieve fast fermentation for a limited risk of microbial contamination.0% sucrose.23% for 4.6°N. 4.5 and 2.48 to 4. 1.5% oat flour and 0. viable cell counts and beta-glucan concentration (B).3 × 108–2.0% was added into the oat mash to increase fermentation rate — results are presented in Fig. combination of aspartame.0% oat flour. 3. Angelov et al. significant differences were observed between cell count increases in the variants. Changes of pH and titratable acidity (A). two variants of oat mashes with the addition of different sweeteners (Var. The initial beta-glucan content in the oat mashes obtained with different flour concentrations was 0.3.5% sucrose such value was registered 0. Results of the experiment are presented in Fig. reaching values of 4. Therefore. 5. At the end of the fermentation. B and Var. The insignificant differences in pH dynamics between the variants with 1. Of the three combinations.0% sucrose and the observed viable cell counts increase support the conclusion that the addition of 1.5 × 1010 cfu ml− 1 at the end of the fermentation. Results showed that sweeteners applied had no effect on the fermentation. 3.0% oat flour.0% sugar added. Acid production was quite slower with 1. 6. Viable cell counts in the three mashes reached 4. and with 1.0% sucrose in the medium and pH 4. Since differences between pH and cell count changes at oat flour concentrations 4.41% for 7. Vinderola et al.0%. 0. Similarly. Changes of pH were similar for all three variants.5% sucrose to the oat mash was not sufficient to provide acceptable sweetness of the oat products. Fig. sucrose — 1.4–1. Effect of sweeteners The addition of 1.5 h later.5%. 3. plantarum A28 starter culture — 5. with final levels of 1.5 and 2. sodium cyclamate and saccharine. fermentation time — 8 h. A laboratory model for the production of an oat-based drink The laboratory model for oat drink production was based on the above estimated conditions: concentration of L. 5.55 at the 6th hour. With 2. respectively.5%. The initial viable cell counts in the three product variants were within 1. Variant A was used as control.5% was preferred in order to obtain higher beta-glucan content in the fermented products.5% sucrose to the oat mash is sufficient for obtaining a probiotic product. .3 × 1010–8.36% for 5. (2002) found no effect of aspartame on the growth of probiotic lactic acid bacteria. TA changed correspondingly. 3. pH 4. Laboratory model for the production of an oat drink. variant C was preferred for application in oat drinks because of the better taste and suitability for the diet of diabetics. Establishment of the oat drink shelf life under refrigerated storage (4–6 °C). register similar cell counts. C) were fermented.0.54 was reached after 8 h. Sucrose concentration Sucrose in concentrations of 1.0 and 5. with the highest of 2.81 log orders for 2.5.1 × 108 cfu ml− 1. which indicated that the starter culture did not ferment beta-glucan. No significant changes of these concentrations were found during fermentation.A.4.

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