黑曲霉酸性脂肪酶Tgl的酶学性质及其酯化能力研究  

Studies on enzymatic characteristics and esterification ability of acid lipase Tgl from Aspergillus niger

作　　者：李文瑶 陈非凡 郑佳 刘怡宁 杜丽平[1] 卢彦坪 马立娟[1] LI Wenyao;CHEN Feifan;ZHENG Jia;LIU Yining;DU Liping;LU Yanping;MA Lijuan(College of Biotechnology,Tianjin University of Science and Technology,Tianjin 300457,China;Yibin Wuliangye Group Co.,Ltd.,Yibin 644000,China)

机构地区：[1]天津科技大学生物工程学院,天津300457 [2]宜宾五粮液股份有限公司,四川宜宾644000

出　　处：《食品与发酵工业》2024年第10期237-243,共7页Food and Fermentation Industries

基　　金：中国轻工业浓香型白酒固态发酵重点实验室开放基金(2021zd012)。

摘　　要：脂肪酶是一种具有水解、酯化、酯交换等多种功能的生物催化剂,为研究脂肪酶的酯化能力,该文克隆表达了来源于黑曲霉的脂肪酶Tgl编码基因,对其进行生物信息学分析,研究该酶的酶学性质及其催化合成脂肪酸乙酯的能力。结果表明,脂肪酶Tgl的最适反应温度为50℃,在50℃下孵育12 h后残余酶活力仍能达到70%左右;最适反应pH值为5.0,在pH 5.0条件下孵育12 h后,其相对酶活力为70%。有机试剂对脂肪酶Tgl均表现出不同程度的抑制作用,其中,乙醇对脂肪酶的抑制作用最为明显,但是在40%的乙醇体系下,Tgl的残余酶活力仍接近80%。20%的乙醇体系中,Tgl催化合成乙酸乙酯的产量最高,为41.62 mg/L,而在60%乙醇体系中催化合成己酸乙酯的产量最高,可达171.08 mg/L。由此可见,黑曲霉酸性脂肪酶Tgl具有较高的催化脂肪酸乙酯合成的能力,在酿造工业中具有很大的应用潜力。Lipase is a biocatalyst with multiple catalytic capacity such as hydrolysis,esterification,and transesterification.To study the esterification ability of lipase,the gene encoding lipase Tgl from Aspergillus niger was cloned and expressed,and the structure,enzymatic characteristics as well as the esterification ability to synthesize fatty acid ethyl ester of Tgl were studied in this work.Results showed that the optimal reaction temperature of Tgl was 50℃,and the residual enzyme activity was about 70%after incubation at 50℃for 12 h.The optimal reaction pH of Tgl was 5.0,and the relative enzyme activity was about 70%after incubation at pH 5.0 for 12 h.Organic reagents showed different degrees of inhibition on Tgl,among which ethanol had the most obvious inhibitory effect on Tgl,but the residual enzyme activity of Tgl was still close to 80%under 40%ethanol system.In 20%ethanol system,the yield of ethyl acetate(41.62 mg/L)catalyzed by Tgl was the highest,while that of ethyl hexanoate was the highest,up to 171.08 mg/L,in 60%ethanol system.In conclusion,acid lipase Tgl from A.niger had high ability to catalyze the synthesis of fatty acid ethyl ester,which would have great application potential in brewing industry.

关 键 词：黑曲霉 脂肪酶 生物信息学分析 酶学性质 酯化能力 

分 类 号：Q936[生物学—微生物学] TQ426.97[化学工程]