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International Journal of Biological Macromolecules 50 (2012) 1008–1010

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International Journal of Biological Macromolecules
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Dehydration of crude protein from Ginkgo biloba L. by microwave freeze drying
Liuping Fan a , Shaodong Ding a , Yuanfa Liu a,∗ , Lianzhong Ai b
a b

State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi 214122, China State Key Laboratory of Dairy Biotechnology, Technology Center, Bright Dairy & Food Co. Ltd., Shanghai 200436, China

a r t i c l e

i n f o

a b s t r a c t
The paper optimized the parameters of microwave freeze drying (MFD) of crude Ginkgo biloba protein (CPG) using response surface methodology (RSM) based on the analysis of its proximate composition. The results showed that coefficients of determination, R2 values for drying time and protein solubility were greater than 0.9500. The drying time and protein solubility of CPG varied curvilinearly with increase of microwave power and pre-freeze temperature, and drying time varied linearly with material thickness. The optimum MFD condition was microwave power of 408–421 W, material thickness of 15 mm and pre-freeze temperature of −20 ◦ C to −21 ◦ C, respectively. © 2012 Elsevier B.V. All rights reserved.

Article history: Received 14 February 2012 Accepted 22 February 2012 Available online xxx Keywords: Ginkgo biloba L. Protein Microwave freeze drying

1. Introduction Numerous techniques can be used to dry solutions of proteins and obtain protein powders with desirable characteristics [1]. However, the most widely used techniques to dehydrate proteins are vacuum freeze drying (FD). The FD dehydration can minimize the losses of active components of foods owing to low temperature and low absolute pressure [2]. In the other hand, FD is also recognized as an expensive technology, because it requires much energy. The costs are 4–8 times higher compared to air drying [3]. There are lots of researches related to reducing FD costs by improving heat and mass transfer [4,5]. Among these improvements, MFD appears to be one of the most promising techniques to accelerate the rate of dehydration and enhance overall quality. When applied to the FD process, microwave energy can penetrate very well into ice and reduces the drying time by as much as 75% [6]. Recently, MFD has been used successfully for drying beef, skim milk, cabbage et al. [7–9]. Ginkgo biloba L. (usually called ginkgo) can date back to 200 million years ago and is considered as a living fossil. The proteins of ginkgo with special biological activity have attracted extensive attention [10–12]. These researches indicated that Ginkgo biloba seed proteins exhibited favorable bioactivity, and could be applied in the food industry as functional additives. However, few of studies have been conducted on the effects of drying methods on the characteristics of Ginkgo seeds protein. The purpose of this paper is to optimize the microwave freeze drying parameters of crude protein powders from Ginkgo biloba.

2. Materials and methods 2.1. Materials The Ginkgo biloba L. cv Dafozhi seeds were purchased from Taixing of Jiangsu province (China). All reagents were of analytical grade and purchased from Sinopharm Chemical Reagent Co., Ltd (China). All results represent the average of duplicate determinations.

2.2. Chemical composition analysis Protein content was calculated from the nitrogen content (%N × 6.25) analyzed by Kjeldahl method. Water and fat content were determined by the official method of AOAC. Total sugar content was examined using phenol–sulfuric acid colorimetric method. The polyphenol content was measured by the Folin–Ciocalteau method.

2.3. Extraction of protein from Ginkgo biloba L. seeds Crude Ginkgo seed protein (CPG) was obtained by an alkaline dissolving and acid precipitating method [13]. The defatted flour of Ginkgo biloba L. was dispersed in distilled water (10%, w/v), and the pH was adjusted to 9.0 using 1 M NaOH. The suspension was stirred for 30 min at room temperature, and then centrifuged at 6000 × g for 15 min. The extracts were combined and acidified to pH 4.4 with 1 M HCl, and then left to stand for 30 min to separate into two layers. The precipitates were recovered by centrifugation, then neutralised by 1.0 M NaOH to pH 7.0 and dialysed in distilled water for 24 h. The neutralised precipitate was dried using MFD.

∗ Corresponding author. Tel.: +86 0510 85876799. E-mail address: (Y. Liu). 0141-8130/$ – see front matter © 2012 Elsevier B.V. All rights reserved. doi:10.1016/j.ijbiomac.2012.02.027

Statistical analysis Data were analysed by using the Statistical Analysis System (SAS. CPG was first frozen for 24 h.2 4. R2 and p values for the response function (Eq.93b 1. The protein suspension was stirred using a magnetic stirrer for 60 min at a speed of 500 rpm and were centrifuged at 6000 × g for 30 min.2.5b 0.05. Proximate composition The proximate composition results of the de-hulled Ginkgo biloba L. Coefficient b0 b1 b2 b3 b11 b22 b33 b12 b13 b23 R2 CV P a b c 2.75 2.15. As a general rule. 3.73b 0.79b −4. Results revealed that the drying time of CPG varied from 4.60 0.1. The coded values of the three independent variables are summarized in Table 1.05) affected by the linear terms of X1 .01 0.75a −8.0003 Y2 (solubility) 66.4 5.9 8. During MFD.01.98c −0.43%. Nanjing Yatai Microwave Power Technology Research Institute. d. The CV of the model was calculated as 6. Effects of MFD on the drying time The drying time (Y1 ) of CPG was significantly (P < 0.15 0.3 48 50 56 55 48 60 59 57 53 52 50 59 67 66 66 Drying time (h) Solubility (%) 2. MFD optimization of CPG A RSM was used to optimize the MFD whilst taking short drying time and retaining high solubility of CPG. material thickness (X2 ) and pre-freeze temperature (X3 ).1 5. The two dependent variables were the drying time (Y1 ) and protein solubility (Y2 ).17 ± 0. d.53 −2. respectively.2 4. which makes Ginkgo biloba a good supplement to protein resource.21 ± 0.33 2.1 10.05. the pressure was maintained at 100 Pa by a vacuum pump.29c −5. Low contents of lipid (5. and the cold trap temperature (−40 to −45 ◦ C) was low enough to condense vapor. The soluble protein content in the supernatant was determined by Kjeldahl method.4 1.9751) indicated that the variables were adequately fitted to the regression equation. The coefficient of variation (CV) is the ratio of the standard error of estimate to the mean value of observed response expressed as a percentage. NC.54b −0.5 4. material thickness (X2 ) and pre-freeze temperature (X3 ).05b −0. Cary.3 10. 2 2 2 Y = b0 + b1 x1 + b2 x2 + b3 x3 + b11¦ x1 + b22 x2 + b33 x3 + b12 x1 x2 + b13 x1 x3 + b23 x2 x3 (1) 2. d. respectively.45 ± 0.13 1. (1)). followed by starch and protein with 71.18%.5. interact terms of X1 X3 and Table 3 Regression coeffcients.13 1. sample. USA). Analyses of variance were performed by the anova procedure. / International Journal of Biological Macromolecules 50 (2012) 1008–1010 Table 1 The coded values of the independent variables.L.9 10.) were found in de-hulled Ginkgo biloba L. Properties analysis of CPG by MFD Drying time was determined according to the moisture content of the samples which dropped to 5% on the wet basis. Y1 (drying time) 5.57 ± 0. total sugar (3. Variable Codes −1 X1 (W) X2 (mm) X3 (◦ C) 350 10 −25 0 400 15 −20 1 450 20 −15 1009 Table 2 The results of response surface analysis of the variation of drying time (Y1 ) and solubility (Y2 ) with microwave power (X1 ).4 h to 11.38 ± 1.4 5. Mean values were considered significantly different when p < 0.27% and 11.9875 and 0.65c 1.b. The protein content is high on a dry matter basis. a model can be considered reasonably reproducible if its CV is not greater than 10% [14].01 Significant at p < 0.. Data were analyzed by the SAS multivariate regression program and could be fitted to the following equation.).b. The coded values of the three independent variables and the results are summarized in Table 2. The high coefficients of determination R2 (0. Process optimization 2.6.9875 6.b. Drying experiments of CPG Experiments were carried out in a lab-scale microwave freeze dryer (YT2S-01. It is a measure of reproducibility of the models. 3. and they found that there is 9–13% crude protein (dry basis) in Ginkgo biloba seed [10].9 5. . The regression coefficients and analysis of variance of CPG are listed in Table 3.) and total phenolic (3. Run Coded variables X1 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 −1 −1 1 1 0 0 0 0 −1 1 −1 1 0 0 0 X2 −1 1 −1 1 −1 −1 1 1 0 0 0 0 0 0 0 X3 0 0 0 0 −1 1 −1 1 −1 −1 1 1 0 0 0 8. For the protein solubility test. cv Dafozhi showed that water was the dominant component (48. solubility of CPG varied from 48% to 67%.4.0. China).001. Before microwave freeze drying (MFD).16% in dry basis.3. X2 and X3 . The probability (p) values of all regression models were less than 0.9 h. The microwave frequency was 2450 MHz and the power could be regulated continually from 0 to 2000 W. CPG of 1% was dispersed in phosphate buffer of pH 7.0017 Significant at p < 0.7. The three independent variables in this experiment were microwave power (X1 ).3 7. Results and discussion 3.0 at room temperature.5 11.36 ± 1.01%. Significant at p < 0. The protein content of sample was in agreement with the reports by Deng et al. Fan et al. SAS Institute Inc.9751 3.89% in wet basis). 3.63b 1.5a −3. version8.28c 0.6% and 3.8 11.

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