鱼鳞胶原肽的分步酶解法制备及其特性研究
发布时间：2018-10-15 11:38
【摘要】：鱼鳞是有鳞鱼类加工过程中产生的副产物,2008年我国淡水鱼总产量1998万吨,其中有鳞鱼1310万吨以上,每年大约可产生65万吨的鱼鳞,除部分以原料形式出口于日本外,大部分未得到很好的开发利用。目前国内已开展鱼鳞酶解工艺研究,且多采用大酶量、长酶解时间及恒定pH等技术来获取较高的得率和水解度,从而导致产物苦味较重、盐分含量过高,在工业化生产中不得不进行脱苦、脱盐处理。本课题以鱼鳞为对象,在分析不同储藏时间及不同鱼种的鱼鳞基本成分变化的基础上,研究了水解条件和预处理方式对鱼鳞酶解效果的影响,评价了三种蛋白酶酶解进程中酶解产物的功能特性,建立了分步酶解法制备高抗氧化活性的鱼鳞胶原肽的生产工艺。主要研究内容和结果如下： (1)鱼鳞原料的确定。测定了储藏0.5～4年的草鱼和鲢鱼鳞及五种淡水鱼(草鱼、鲫鱼、鲤鱼、鲢鱼和鳊鱼)鱼鳞的基本成分。储藏时间对鱼鳞的胶原蛋白含量影响不大,草鱼、鳊鱼及鲢鱼鳞中胶原蛋白含量较高,且草鱼鳞片较大便于收集,故选择存放0.5年的干草鱼鳞为研究对象进行后续酶解实验。 (2)水解条件的确定。起始pH值、热处理温度、热处理时间、底物浓度、加酶量对草鱼鳞水解产物的水解度和氮收率均有显著影响(p0.01)。碱性蛋白酶水解草鱼鳞的适宜条件为：料液比1：17,热处理温度85℃,热处理时间1.5h,起始pH值8.0～8.5,加酶量420u/gpro,在60℃水解4.5h。中性蛋白酶和复合蛋白酶水解草鱼鳞的适宜条件为：料液比1：17,热处理温度85℃,热处理时间1.5h,起始pH值7.0～7.5,加酶量450u/gpro,在50℃水解4.5h。 (3)草鱼鳞的预处理方式的确定。预处理对草鱼鳞酶解产物的氮收率、水解度和氨基酸生成量有显著影响(p0.01)。在粉碎-热处理、不粉碎-热处理、不粉碎-不热处理等三种处理方式中,粉碎-热处理的草鱼鳞的酶解产物的氮收率、水解度和氨基酸生成量均最高。预处理对鱼鳞的复合蛋白酶和中性蛋白酶酶解产物抗氧化活性影响不大,而对鱼鳞的碱性蛋白酶酶解产物的抗氧化活性有明显影响。对于碱性蛋白酶酶解产物而言,不粉碎-不热处理鱼鳞的酶解产物的抗氧化活性最强(DPPH清除率的IC50值最小),但其氮收率仅71%,远低于经粉碎和加热处理鱼鳞的酶解产物的氮收率的(99%左右)。预处理方式对酶解产物分子量分布的影响不明显。 (4)酶解进程中产物特性的研究。三种蛋白酶水解产物的氮收率、水解度、游离氨基酸、乳化活力指数(ESI)及抗氧化活性随酶解时间的延长其变化规律相似。酶解时间为0～4h,其氮收率、水解度、游离氨基酸及DPPH自由基清除率呈增大趋势,ESI和分子量呈减小趋势。酶解超过4h后继续延长酶解时间,碱性蛋白酶、复合蛋白酶和中性蛋白酶水解液氮收率分别维持在97%、97%、90%左右,而DPPH自由基清除率呈下降趋势,产物分子量分布无明显变化。 (5)两步酶解法制备抗氧化肽。碱性蛋白酶与风味蛋白酶分步酶解草鱼鳞的水解产物水解度和氨基酸生成量最高,当总酶活为840 u/gpro时,其水解度和氨基酸生成量分别为15%和130mg/g鱼鳞。水解产物的DPPH自由基清除率随总酶活的增加而升高,当总酶活高于525 u/gpro时,碱性蛋白酶与复合蛋白酶分步酶解产物的DPPH自由基清除率最高。在总酶活为840u/gpro时,碱性蛋白酶与复合蛋白酶分步酶解产物的平均分子量最小,其中分子量高于9952Da的多肽含量仅占14%,且5946Da以下的多肽占多肽总量的1/2左右。
[Abstract]:Fish scale is the by-product produced during the process of fish scale fish. In 2008, the total output of freshwater fish in China was 1998 million tons. Among them, there were more than 13.1 million tons of scale fish, about 65 million tons of fish scale can be produced every year, and most of the fish scales were exported to Japan in the form of raw materials, and most of them were not well developed and developed. At present, fish scale enzymatic hydrolysis technology has been carried out in China, and the high yield and degree of hydrolysis can be obtained by adopting the technology of large enzyme amount, long enzymolysis time and constant pH value, so that the bitter taste of the product is heavy, the content of salt is too high, and the debittering and desalting treatment of the fish scale enzyme solution has to be carried out in industrial production. Based on the analysis of the changes of fish scales in different storage times and different species, the effects of hydrolysis conditions and pretreatment on the enzymatic hydrolysis of fish scales were studied. A process for preparing fish scale collagen peptide with high antioxidant activity by stepwise enzymatic hydrolysis was established. The main contents and results are as follows: (1) fish scale raw material Determination of fish scales of grass carp and fish scales and five freshwater fishes (grass carp, Carassius auratus, Cyprinus carpio, sturgeon and sturgeon) for 0. 5 ~ 4 years were determined. The content of collagen in fish scale was not big, but the content of collagen in grass carp, sturgeon and fish scale was higher, and the fish scale of grass was more convenient to collect, so it was chosen to store 0. 5 years' hay scale was the research object for follow-up enzymolysis. Experiment. (2) Hydrolysis The conditions were determined. The initial pH value, heat treatment temperature, heat treatment time, substrate concentration, enzyme addition amount had significant influence on the hydrolysis degree and the nitrogen yield of the fish scale hydrolysate (p The optimum conditions are: the ratio of the liquid to the fish scale is 1: 17, the heat treatment temperature is 85.d egree. C., the heat treatment time is 1. 5h, the initial pH value is 8.0-8.5, the enzyme amount is 420u/ gro, The optimum conditions were: the ratio of liquid to liquid was 1: 17, the heat treatment temperature was 85 鈩，
本文编号：2272432
[Abstract]:Fish scale is the by-product produced during the process of fish scale fish. In 2008, the total output of freshwater fish in China was 1998 million tons. Among them, there were more than 13.1 million tons of scale fish, about 65 million tons of fish scale can be produced every year, and most of the fish scales were exported to Japan in the form of raw materials, and most of them were not well developed and developed. At present, fish scale enzymatic hydrolysis technology has been carried out in China, and the high yield and degree of hydrolysis can be obtained by adopting the technology of large enzyme amount, long enzymolysis time and constant pH value, so that the bitter taste of the product is heavy, the content of salt is too high, and the debittering and desalting treatment of the fish scale enzyme solution has to be carried out in industrial production. Based on the analysis of the changes of fish scales in different storage times and different species, the effects of hydrolysis conditions and pretreatment on the enzymatic hydrolysis of fish scales were studied. A process for preparing fish scale collagen peptide with high antioxidant activity by stepwise enzymatic hydrolysis was established. The main contents and results are as follows: (1) fish scale raw material Determination of fish scales of grass carp and fish scales and five freshwater fishes (grass carp, Carassius auratus, Cyprinus carpio, sturgeon and sturgeon) for 0. 5 ~ 4 years were determined. The content of collagen in fish scale was not big, but the content of collagen in grass carp, sturgeon and fish scale was higher, and the fish scale of grass was more convenient to collect, so it was chosen to store 0. 5 years' hay scale was the research object for follow-up enzymolysis. Experiment. (2) Hydrolysis The conditions were determined. The initial pH value, heat treatment temperature, heat treatment time, substrate concentration, enzyme addition amount had significant influence on the hydrolysis degree and the nitrogen yield of the fish scale hydrolysate (p The optimum conditions are: the ratio of the liquid to the fish scale is 1: 17, the heat treatment temperature is 85.d egree. C., the heat treatment time is 1. 5h, the initial pH value is 8.0-8.5, the enzyme amount is 420u/ gro, The optimum conditions were: the ratio of liquid to liquid was 1: 17, the heat treatment temperature was 85 鈩，
本文编号：2272432
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