食品与发酵工业 >

2025 , Vol. 51 >Issue 1: 41 - 46

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.038635

研究报告

疏棉状嗜热丝孢菌脂肪酶LipTL的酶学性质及脱胶应用研究

  • 邵化 ,
  • 单明明 ,
  • 王琼波 ,
  • 徐瑞敏 ,
  • 陈曦 ,
  • 魏涛
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  • 1(郑州轻工业大学 食品与生物工程学院,河南 郑州,450001)
    2(漯河医学高等专科学校 食品营养系,河南 漯河,462002)
第一作者:博士,讲师(魏涛教授为通信作者,E-mail:weit8008@zzuli.edu.cn)

收稿日期: 2024-01-22

  修回日期: 2024-03-21

  网络出版日期: 2025-01-23

基金资助

中原科技创新人才项目(224200510017);河南省重点研发专项(231111111700)

收起

Expression and characterization of the lipase LipTL from Thermomyces lanuginosus and its application in oil degumming

  • SHAO Hua ,
  • SHAN Mingming ,
  • WANG Qiongbo ,
  • XU Ruimin ,
  • CHEN Xi ,
  • WEI Tao
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  • 1(College of Food and Bioengineering, Zhengzhou University of Light Industry, Zhengzhou 450001, China)
    2(Department of Food and Nutrition, Luohe Medical College, Luohe 462002, China)

Received date: 2024-01-22

  Revised date: 2024-03-21

  Online published: 2025-01-23

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摘要

酶法脱胶技术在油脂精炼行业应用潜力巨大,磷脂酶是酶法脱胶技术中的主要用酶,但因现有磷脂酶资源在底物选择性、温度稳定性方面难以满足工业脱胶工艺的要求,限制了磷脂酶在油脂精炼行业中的大规模应用。研究表明,来自疏棉状嗜热丝孢菌(Thermomyces lanuginosus)的脂肪酶(LipTL)具有水解磷脂的功能。该研究利用大肠杆菌构建LipTL的基因工程菌,对LipTL进行异源表达,探究LipTL的酶学性质和油脂脱胶效果。序列比对显示,该酶与磷脂酶A1具有高度相似性;SDS-PAGE结果表明,LipTL分子质量约为30 kDa;LipTL的最适温度为55 ℃、最适pH值为9.0,在55 ℃条件下处理2 h仍能保持90%以上的相对活力。LipTL在脱胶反应6 h后,将大豆油和花生油的残磷量分别降至5.2、4.9 mg/kg,具有良好的脱胶效果。该研究结果证实该脂肪酶可用于油脂脱胶,不仅为酶法脱胶提供新的酶源,也为该酶的后续改造提供了研究基础。

关键词： 疏棉状嗜热丝孢菌; 脂肪酶; 酶法脱胶; 同源建模; 酶学性质

本文引用格式

邵化 , 单明明 , 王琼波 , 徐瑞敏 , 陈曦 , 魏涛 . 疏棉状嗜热丝孢菌脂肪酶LipTL的酶学性质及脱胶应用研究[J]. 食品与发酵工业, 2025 , 51(1) : 41 -46 . DOI: 10.13995/j.cnki.11-1802/ts.038635

Abstract

Enzymatic degumming technology has great potential application in oil refining industry.Phospholipases are the major enzymes that applied in oil degumming.However, the existing phospholipases are unable to meet the requirements of industrial degumming processes in terms of substrate selectivity and temperature stability, which limits the large-scale application of phospholipases in the oil refining industry.Researches have revealed that the lipase (LipTL) from Thermomyces lanuginosus has ability of hydrolyzing phospholipids.In this study, this lipase LipTL gene was heterologously expressed in Escherichia coli, then the recombinant LipTL was purified and characterized, and its application in oil degumming was also studied.Sequence alignment exhibited that LipTL showed high similarity with phospholipase A1;SDS-PAGE revealed that the molecular weight of LipTL was approximately 30 kDa;The optimal temperature and pH for LipTL were 55 ℃ and 9.0 respectively, and it could maintain over 90% relative activity after 2 hours of treatment at 55 ℃.After 6 hours of degumming reaction, LipTL reduced the residual phosphorus content of soybean oil and peanut oil to 5.2 mg/kg and 4.9 mg/kg, respectively.This study confirms that the lipase LipTL can be applied to oil degumming, which not only provides a new enzyme source for enzymatic degumming, but also lays a good foundation for the subsequent modification of this enzyme.

Key words： Thermomyces lanuginosus; lipase; enzymatic degumming; homology modeling; enzymatic properties

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