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1 7 3 Vol. 7 No Journal of Food Safety and Quality Mar., 26, 田景民 2, 陈贵林 (., 2; 2., ; 3., 2), 3* 摘要 : 目的, 方法,, ph 结果 : 88 min 6 ph 5: ml/g : > >ph>, Y= A.33B+3.49C+.63D.56A 2.97B 2 +.9C 2.39D AB.25AC AD BC BD.CD, 7 结论关键词 : ; ; ; Optimization of extraction processing for soluble protein of seed residue of sea buckthorn using the response surface analysis and amino acid composition analysis TIAN Jing-Min, 2, 3*, CHEN Gui-Lin (. College of Life Sciences, Inner Mongolia University, Hohhot 2, China; 2. College of Pharmacy, Inner Mongolia Medical University, Hohhot, China; 3. The Good Agriculture Practice Engineering Technology Research Center of Chinese and Mongolian Medicine in Inner Mongolia, Hohhot 2, China) ABSTRACT: Objective To optimize the extraction processing of water-soluble protein in sea buckthorn seed by response surface method and analyze the amino acid composition of the water-soluble protein. Methods The effects of the ratio of liquid, extraction temperature, ph and extraction time and their interactions on the protein yield were researched based on the single-factor test and the amino acid composition of the water-soluble protein was analyzed. Results The optimum extraction conditions were as follows: extraction time of 88 min, extraction temperature of 6, ph, liquid-to-solid ratio 5: ml/g. The influence order of factors to protein extracting was: extraction time>extraction temperature>ph>liquid-to-solid 基金项目 : (2) Fund: Supported by the Inner Mongolia Science and Technology Innovation to Guide the Reward (2) * 通讯作者 :,,, chengl@life.imu.edu.cn *Corresponding author: CHEN Gui-Lin, Professor, College of Life Science, Inner Mongolia University, No.235 West University Street, Hohhot 2, China. chengl@life.imu.edu.cn

2 322 7 ratio. The mathematic model of the quadratic polynomial regression equation was Y= A.33B+ 3.49C+.63D.56A 2.97B 2 +.9C 2.39D AB.25AC AD BC BD.CD, which was obtained through the response surface method and could exactly predict the yield of protein. Water-soluble protein was made up of 7 kinds of amino acids. Conclusion This study will provide theoretical basis for the extraction and the application of the water-soluble protein of seed residue of sea buckthorn. KEY WORDS: sea buckthorn; response surface methodology; water-soluble protein; composition of amino acid 引言,,,,,, [], 26.3, 2%~25%, 8, 8,, % [2],,,, [3,4],, ph,, 2 材料与方法 2. 材料 : CO 2,, 9%, 2.2 仪器与试剂 98-Ⅱ-B (5 ml)( ); SHZ-CA ( ); AL24 ( - (METTLER TOLEDO) ( ) ); JZ74 ( ); ( ); (, ); (, ) 2.3 实验方法 [5-2] 2.3. 沙棘籽渣水溶性蛋白质最佳提取条件的确定 ph, 3, 5,, Design-Expert 8., 4 3,, 沙棘籽渣水溶性蛋白质含量的测定, [3,4] A%=W 2 *V/W *M % A: ; V: ; W : ; W 2 : ; M: 沙棘籽渣水溶性蛋白质的氨基酸分析 GB/T [5] 2.4 数据处理 Design-Expert 8. Table 表响应面试验因素水平 Factors and levels in response surface design A (ml/g) B ( ) C ph D (min) : : : 3

3 3, : 结果与分析 3. 单因素试验结果与分析 3.. 液料比对沙棘籽渣水溶性蛋白质提取率的影响 ph min, (:)~(3:) [2,3],, ; 2:,,,, 2: 3..2 提取温度对沙棘籽渣水溶性蛋白质提取率的影响 ph9 2: 6 min, 2~ [2,3] 2,, 2~6, 7,,,,, ph 值对沙棘籽渣水溶性蛋白质提取率的影响 7 2: 6 min, ph 5~3 [2,3] 3, ph,, ph 3~5, ph,, ph,,,,,, ph Fig. (n=5) Effect of liquid to solid ratio on the extraction rate of soluble protein of seed residue of sea buckthorn (n=5) Fig. 2 2 (n=5) Effect of extraction temperature on the extraction rate of soluble protein of seed residue of sea buckthorn (n=5)

4 提取时间对沙棘籽渣水溶性蛋白质提取率的影响 7 2: ph, 2~ min [2,3] 4,,, 8 min,,,, 8 min 3.2 响应面法优化试验 3.2. 响应面回归模型的建立与分析 ph 4,, 2 Design Expert, 3 : (%)Y= A.33B+3.49C+.63D.56A 2.97B 2 +.9C 2.39D AB.25AC AD BC BD-.CD 3, (P<.), (P=.986>.5) A B C D (P<.), A 2 B 2 C 2 D 2 CD (P<.5); AB AC AD BC BD (P>.5) Fig. 3 3 ph (n=5) Effect of extraction ph on the extraction rate of soluble protein of seed residue of sea buckthorn (n=5) Fig. 4 4 (n=5) Effect of extraction time on the extraction rate of soluble protein of seed residue of sea buckthorn (n=5)

5 3, : 325 Table 2 表 2 响应面试验设计及结果 Experimental design and results for response surface analysis A (ml/g) B ( ) C ph D (min) % Table 3 表 3 方差分析 Analysis of variance for the fitted regression model F P <. ** A <. ** B <. ** C <. ** D <. ** AB

6 326 7 续表 3 F P AC AD.... BC.... BD.... CD * A <. ** B <. ** C <. ** D <. ** R 2. R 2 Adj. S/N * ; ** F, D( )>A( )>B( )>C(pH) 两因素间交互作用的分析 5a~5f ( 5),,,, ;,,,, ;,,,,, 最佳工艺条件的预测与验证, : 88.9 min, 4.84 ml/g, 59.38, ph.4, %, : 88 min, 5: ml/g, 6, ph 3, 38.6 %,, 3.3 沙棘籽渣水溶性蛋白质氨基酸组成分析 4, mg/ mg

7 第3期田景民, 等: 响应面法优化沙棘籽渣水溶性蛋白质提取工艺研究及其氨基酸组成分析 327 5a Effect of extraction temperature and liquid to solid ratio on the extraction rate of soluble protein of seed residue of sea buckthorn 5b Effect of extraction ph and liquid to solid ratio on the extraction rate of soluble protein of seed residue of sea buckthorn 5c 5d Effect of extraction temperature and extraction ph on the extraction rate of soluble protein of seed residue of sea buckthorn Effect of extraction time and liquid to solid ratio on the extractionrate of soluble protein of seed residue of sea buckthorn 5e Effect of extraction temperature and extraction time on the extraction rate of soluble protein of seed residue of sea buckthorn Fig. 5 5f Effect of extraction time and extraction ph on the extraction rate of soluble protein of seed residue of sea buckthorn 图 5 两因素相互作用对沙棘籽渣水溶性蛋白质提取率的响应面和等高线图 Response surface and contour plots for the effect of operating parameters on the extraction rate of soluble protein of seed residue of sea buckthorn

8 328 7 Table 4 表 4 沙棘籽渣水溶性蛋白质氨基酸组成分析 Analysis of amino acids for the soluble protein of seed residue of sea buckthorn (mg/mg) 讨论 2%~25% [4], 38.6% 7, 22.64%, 6 [3,4],,, [6,7], 参考文献 [],,. : [J]., 27, 9(3): Ma J, Ge X, Chang ZY. Protein function studies: history, current status and future trends [J]. Chin Bull Life Sci, 27, 9(3): [2],,. [J]., 24, 39(9): Liu Y, Lian YS, Wang YL. Review of research and development and significant effect of sea buckthorn [J]. China J Chin Mat Med, 24, 39(9): [3],,. [J]., 28, 22(8): Zhang WB, Ku EB, Zhang Y. Optimization of extraction process of sea buckthorn Xinjiang and amino acid composition analysis [J]. Cereals Oils Proc, 28, 22(8): [4],,. [J]., 22, 37(9): Cui M, Ling MS, Sun MK. Extraction of sea buckthorn seed protein by alkaline extraction and enzymatic hydrolysis [J].

9 3, : 329 China Oils Fats, 22, 37(9): [5],,. [J]., 24, 5(5): Jiang YT, Zhao Y, Cai EB. Extraction process optimization of water-soluble protein from Ganoderma lucidum [J]. J Food Saf Qual, 24, 5(5): [6],. [J]., 24, 5(9): Tian JM, Chen GL. Optimizaton of extraction of total flavones from residue of Hippophae rhamnoides L. using response surface methodology and their antioxidant activity [J]. J Food Saf Qual, 24, 5(9): [7],,. [J]., 25, 2(2): 6 9. Liu C, Yan MM, Shao S. Optimization of extraction process of water-soluble protein from Schisandrae Chinensis Fructus [J]. Chin J Exper Tradit Med Form, 25, 2(2): 6 9. [8]. [J]., 29, 6(6): Zhao CW. Study on extraction of separate protein of sea buckthorn [J]. Food Sci Technol, 29, 6(6): [9] Jiang P, Kang J, Wang L. Study on comprehensive utilization of sea buckthorn pulp in Xinjiang [J]. Food Ferment Ind, 2, 37(5): [] Chen T, Wang CQ, Li XF. Antibacterial actions of enzymolysis polypeptide of seabuckthorn seed in vivo and in vitro [J]. J Food Sci 25, 36(): [] PB Tirupathi Pichiah, Hye-Jung Moon, Jeong-Eun Park, et a. Ethanolic extract of sea buckthorn (Sea buckthorn L.)preve nts high-fat diet-induced obesity in mice through down regulation of adipogenic and lipogenic gene expression [J]. Nutr Res, 22, 32: [2],,. [J]., 22, 33(6): Zeng XF, Yu X, Lin HW. Optimize the extraction technology of Tephrosia candida protein by response surface methodology [J]. Food Res Dev, 22, 33(6): [3] ( ) [M]. :, 25. The State Pharmacopoeia Commission [M]. Beijing: China Medicine Science and Technology Press, 25. [4] GB [S]. GB National food safety standards-determination of protein in foods [S].. [5] GB/T [S]. GB/T Determination of amino acids in food [S]. [6],,. [J]., 2, 26(6): Huang P, Su N, Wang CT. Study on preparation and efficacy of sea buckthorn polypeptide [J]. Food Mach, 2, 26(6): [7]. [J]., 24, 29(4): Wang MG. Antioxidant effect of protease hydrolysate of sea buckthom seed [J]. J Chin Cereals Oils Assoc, 24, 29(4): 作者简介 ( 责任编辑 : 杨翠娜 ) 田景民, 讲师, 博士研究生, 主要研究方向为药用植物化学 tianjingminyaoxue@63.com 陈贵林, 教授, 博士生导师, 主要研究方向为药用植物化学 chengl@life.imu.edu.cn

H 2 SO ml ml 1. 0 ml C 4. 0 ml - 30 min 490 nm 0 ~ 100 μg /ml Zhao = VρN 100% 1 m V ml ρ g

H 2 SO ml ml 1. 0 ml C 4. 0 ml - 30 min 490 nm 0 ~ 100 μg /ml Zhao = VρN 100% 1 m V ml ρ g 16 6 2018 11 Chinese Journal of Bioprocess Engineering Vol. 16 No. 6 Nov. 2018 doi 10. 3969 /j. issn. 1672-3678. 2018. 06. 004 1 2 1 1 1. 330200 2. 330006 4 Box-Behnken 1 25 g /ml 121 92 min 2 6. 284%