鱼油酶法水解及酯交换反应中脂肪酶活力检测方法的研究

    Methods for detecting lipase activity on hydrolysis and transesterification of fish oil

    • 摘要: 利用脂肪酶进行酶法酯交换反应以提高鱼油甘油三酯原料的ω-3功能性脂肪酸含量,是近年来食品和饲料行业的研究热点。准确测定酶活力对脂肪酶在鱼油产业上的应用具有重要意义。本文以8种不同商品化的脂肪酶为原料,分别从酶的水解活力和酯交换活力两个方面研究脂肪酶活力检测方法,并应用于鱼油水解反应和酯交换反应。结果显示,以酶活力测定的标准偏差值为指标,确定了以聚乙烯醇-橄榄油乳化法测定脂肪酶水解活力和正辛酸-三油酸甘油酯交换法测定酯交换活力的方法;水解酶活力和酯交换酶活力拟合的线性模型、幂模型和指数函数模型的相关系数分别是0.07、0.15和0.20,表明两者无显著相关,不能相互预测。上述两种酶活检测方法在分别用于筛选高酶活的鱼油水解和酯交换反应所需的脂肪酶时,脂肪酶L5的鱼油水解产物酸价达到137.61 mg KOH/g,固定化酶L8的酯交换甘油酯产物EPA和DHA总含量为55.38%。

       

      Abstract: The use of lipase in enzymatic transesterification to improve the content of omega-3 functional fatty acids in fish oil triglycerides has been a hot topic in food and feed industry in recent years.Accurate determination of the enzyme activity is of great significance to the application of lipase in fish oil industry.With 8 different commercial lipases as raw materials,the lipase activity detection method was studied from two aspects including the enzymatic hydrolysis activity and transesterification activity,and then applied to fish oil hydrolysis reaction and transesterification reaction.When the standard deviation of enzyme activity determination was used as an indicator,the results showed that polyvinyl alcohol-olive oil emulsification method was more applicable for hydrolytic activity than the p-nitrophenol method,while the octanoic acid-glycerol trioleate transesterification method was more applicable for transesterification activity than the p-nitrophenol method.The correlation coefficients of the lipase transesterification activity and the hydrolytic activity,fitted by linear model,power model and exponential model,were 0.07,0.15 and 0.20,respectively,showing no significant correlation between these two activities and they cannot be predicted by each other.When the two lipase activity detection methods were used to screen high-enzymatic fish oil hydrolysis and transesterification lipases,the acid value of the fish oil hydrolysate of lipase L5 was 137.61 mg KOH/g,and the total contents of EPA and DHA from transesterification products of immobilized enzyme L8 was 55.38%.This study provides a more accurate determination of enzyme activity for the application of lipase in fish oil industry.

       

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