碱性蛋白酶水解苜蓿叶蛋白的工艺优化
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摘要
以苜蓿叶蛋白粉为原料,利用碱性蛋白酶水解制备寡肽,研究底物浓度、加酶量、水解温度和反应体系pH四因素对酶解效率和抗氧化活性的影响。在单因素试验基础上,通过正交试验确定苜蓿叶蛋白的最佳水解工艺条件:温度为50℃,pH为9.0,酶加量为3 000 U/g,底物浓度为3%。在该条件下,蛋白水解度(DH)为19.04%,水解液的可溶性蛋白含量为52.01 mg/mL,寡肽的平均肽链长度为5.25。不同水解度的酶解物对二苯代苦味肼基自由基(DPPH·)清除率较高,酶解物的水解度与DPPH·清除率之间不存在线性关系。
Took alfalfa leaf protein as experiment material, the oligopeptide was produced through hydrolysis by alcalase. The impact effects of four factors including substrate concentration, adding amount of enzyme, hydrolysis temperature and pH value on hydrolysis effi ciency and antioxidant activity were studied. Through orthogonal test, the results showed the optimal conditions of hydrolyzing alfalfa leaf protein were as follows: hydrolyzing temperature was 50 ℃, pH value was 9.0, adding amount of enzyme was 3 000 U/g, substrate concentration was 3%. Under those conditions, degree of hydrolysis(DH) of alfalfa leaf protein was 19.04%, the content of soluble protein of hydrolysates was 52.01 mg/mL, average chain length of the oligopeptide was 5.25. Scavenging activity of alfalfa leaf protein hydrolysates with different DH against DPPH radical is higher, there was no linear relationship between degree of hydrolysis and scavenging activity of alfalfa leaf protein hydrolysates against DPPH radical.
Took alfalfa leaf protein as experiment material, the oligopeptide was produced through hydrolysis by alcalase. The impact effects of four factors including substrate concentration, adding amount of enzyme, hydrolysis temperature and pH value on hydrolysis effi ciency and antioxidant activity were studied. Through orthogonal test, the results showed the optimal conditions of hydrolyzing alfalfa leaf protein were as follows: hydrolyzing temperature was 50 ℃, pH value was 9.0, adding amount of enzyme was 3 000 U/g, substrate concentration was 3%. Under those conditions, degree of hydrolysis(DH) of alfalfa leaf protein was 19.04%, the content of soluble protein of hydrolysates was 52.01 mg/mL, average chain length of the oligopeptide was 5.25. Scavenging activity of alfalfa leaf protein hydrolysates with different DH against DPPH radical is higher, there was no linear relationship between degree of hydrolysis and scavenging activity of alfalfa leaf protein hydrolysates against DPPH radical.
引文
[1]杨春波.苜蓿叶蛋白的提取与应用研究[D].江南大学,2007.
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[2]SAKO K,IMAMURA Y,NISHIMATA H,et al.Branchedchain amino acids supplements in the late evening decrease the frequency of muscle cramps with advanced hepatic cirrhoss[J].Hepatol Res,2003,26(4):327-329.
[3]陈毓荃,马静芳,文建雷,等.生物化学实验方法和技术[M].北京:科学出版社,2002.
[4]张水华.食品分析[M].北京:中国轻工业出版社,2007:159-160.
[5]SHIMADA K,FUJIKAWA K,YAHARA K,et al.Antioxidative properties of xanthum on the antioxidation of soybean oil in cyclodextrin emulsion[J].Journal of Agricultural and Food Chemistry,1992,40(6):945-948.
[6]熊燕飞,王惠平,陈怀新,等.酶法水解蝉蛹蛋白制备多肽的工艺研究[J].湖北大学学报:自然科学版,1999(9):287-288.