产低温脂肪酶菌株鉴定、发酵条件优化及酶学性质分析  

Identification of A Cold-active Lipase Producing Strain,Optimization of Fermentation Conditions and Analysis of Enzymatic Properties

作　　者：刘思远 申东晨 刘峥 鲁丽颖 徐恒 董爱荣[1] LIU Siyuan;SHEN Dongchen;LIU Zheng;LU Liying;XU Heng;DONG Airong(College of Forestry,Northeast Forestry University,Harbin 150040,China)

机构地区：[1]东北林业大学林学院,黑龙江哈尔滨150040

出　　处：《食品工业科技》2023年第20期116-125,共10页Science and Technology of Food Industry

基　　金：国家自然科学基金项目(31670494)。

摘　　要：为筛选高产低温脂肪酶的菌株,并对产酶条件进行优化,同时为脂肪酶的工业化开发提供生产资料。从黑龙江省漠河县土壤样品中筛选出一株产低温脂肪酶菌株,通过形态学鉴定、生理生化实验及分子生物学鉴定,确定该菌株为普城沙雷氏菌(Serratiaplymuthica)。通过单因素实验,探究温度、pH、装液量、接种量、碳源、氮源、金属离子、诱导剂等不同因素对菌株产酶的影响,通过Plackett-Burman实验,爬坡试验及响应曲面设计,优化橄榄油、蛋白胨、装液量等因素的添加量。结果表明:该菌株最优产酶条件为20℃、pH7.5、装液量42 mL、接种量0.5%、20 g/L麦芽糖、14 g/L蛋白胨、0.5 g/L的MgSO_(4)·7H_(2)O及46 mL/L橄榄油。此优化条件下,脂肪酶活为98.05 U/mL,是优化前的5.85倍。酶学性质结果表明,该脂肪酶最适温度为30℃,属于低温脂肪酶,最适反应pH为7,Mg^(2+)明显可以促进酶活。有机溶剂中甲醇和乙醇明显抑制了酶活性,而正己烷明显可以促进酶活性。该结论可为微生物资源开发利用,工业生产低温脂肪酶提供一定的理论依据及方法指导。To screen strains with high production of cold-active lipase and optimize enzyme production conditions,as well as to provide production information for the industrial development of lipase,a cold-active lipase producing strain was screened from soil samples in Mohe County,Heilongjiang Province,and identified as Serratia plymuthica by morphological identification,physiological and biochemical experiments and molecular biology.The effects of different factors such as temperature,pH,loading volume,inoculum,carbon source,nitrogen source,metal ion and inducer on the enzyme production of the strain were investigated by single-factor experiments,as well as the optimization of the addition of olive oil,peptone and loading volume by Plackett-Burman experiment,hill climbing test and response surface design.The results showed that the optimal enzyme production conditions were 20℃,pH7.5,loading volume 42 mL,inoculum 0.5%,20 g/L maltose,14 g/L peptone,0.5 g/L MgSO_(4)·7H_(2)O and 46 mL/L olive oil.The lipase activity under this optimized condition was 98.05 U/mL,which was 5.85 times higher than that before optimization.The results of enzymatic properties showed that the optimum temperature of this lipase was 30℃,which was a low temperature lipase,and the optimum reaction pH was 7.Mg^(2+)could obviously promote the enzyme activity.The organic solvents methanol and ethanol obviously inhibited the enzyme activity,while n-hexane could obviously promote the enzyme activity.The conclusion can provide some theoretical basis and methodological guidance for the development and utilization of microbial resources and industrial production of low-temperature lipase.

关 键 词：低温脂肪酶 普城沙雷氏菌 发酵条件优化 酶学性质 

分 类 号：TQ925[轻工技术与工程—发酵工程]