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鹿茸胶原多肽复合酶水解工艺研究
李 冰1, 李 梁1, 王露露,等1
吉林农业大学 中药材学院
摘要:
【目的】对鹿茸胶原多肽进行复合酶水解,优化水解工艺,并测定水解液分子量分布。【方法】以碱性蛋白酶和胰蛋白酶为供试酶类,选用酶解时间、pH值、酶解温度、加酶量为影响因素,采用响应面试验设计优化单酶水解条件,根据响应面试验所得到的单酶水解的最适条件,确定双酶复合水解方案。通过 Sephadex G25测定鹿茸胶原多肽的分子量分布。【结果】碱性蛋白酶最优水解条件为:酶解时间3.6 h,pH值10.50,酶解温度55 ℃,加酶量4%,水解液的水解度为22.03%;胰蛋白酶最优水解条件为:酶解时间3.2 h,pH值8.00,酶解温度50 ℃,加酶量4.4%,水解液的水解度为15.81%。复合酶水解条件:酶解温度为52.5 ℃,调节pH值为10.50,加入4%的碱性蛋白酶酶解3.6 h;调节pH值至8.00,加入4.4%的胰蛋白酶酶解3.2 h,所得水解液的水解度可达33.42%。复合酶水解胶原多肽的分子量分布在400~1 400 u。【结论】确定了鹿茸胶原多肽碱性蛋白酶和胰蛋白酶复合水解的工艺条件。
关键词:  鹿茸  响应面试验  复合酶水解  胶原多肽
DOI:
分类号:
基金项目:吉林省科技厅发展计划项目(20140204063YY,20130413044GH,20120246)
Composite enzymatic hydrolysis of collagen peptide from antler
LI Bing,LI Liang,WANG Lulu,et al
Abstract:
【Objective】Hydrolysis of collagen peptide of antler by composite enzymes was optimized and the molecular weight (MW) distribution of hydrolysates was investigated.【Method】Response surface design experiment was applied to optimize the hydrolysis conditions (including hydrolysis time,pH value,hydrolysis temperature and enzyme loading) for alkaline protease and trypsin to determine the optimal composite enzymatic hydrolysis of collagen peptide from antler.The molecular weight distribution of hydrolysates was also determined using Sephadex G25 partition.【Result】The optimal conditions for alkaline protease were pH 10.50,temperature 55 ℃,and enzyme loading 4% for 3.6 h with the degree of hydrolysate (DH) of 22.03%.The optimal conditions for trypsin were pH 8.00,temperature 50 ℃,and enzyme loading 4.4% for 3.2 h with DH of 15.81%.The optimum parameters of composite enzyme hydrolysis were hydrolyzed 4% alkaline protease at 52.5 ℃ and pH 10.50 for 3.6 h before hydrolyzing 4.4% trypsin at pH 8.00 for 3.2 h.The obtained DH was 33.42%.The molecular weight distribution of collagen peptide from antler ranged from 400 to 1 400 u.【Conclusion】This study determined the optimal conditions for composite enzymatic hydrolysis of collagen peptide from antler by alkaline protease and trypsin.
Key words:  velvet antler  response surface design experiment  composite enzymatic hydrolysis  collagen peptide

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