植物乳杆菌漆酶的酶学性质及其对生物胺的降解作用

doi:10.7506/spkx1002-6630-20221025-258

食品科学 ›› 2023, Vol. 44 ›› Issue (12): 157-163.doi: 10.7506/spkx1002-6630-20221025-258

植物乳杆菌漆酶的酶学性质及其对生物胺的降解作用

曹玉莹，王爽，刘怡宁，高熳熳，杜丽平，马立娟，张惠玲，田晓菊，杨伟明

（1.天津科技大学生物工程学院，天津 300457；2.天津科技大学工业发酵微生物教育部重点实验室，天津市工业微生物重点实验室，天津 300457；3.宁夏大学食品与葡萄酒学院，宁夏银川 750021；4.宁夏志辉源石葡萄酒庄有限公司，宁夏银川 750026）

出版日期:2023-06-25 发布日期:2023-06-30
基金资助:
宁夏回族自治区重点研发计划项目（2020BFH02006）

Enzymatic Characteristics of Laccase from Lactobacillus plantarum and Its Effect on Degradation of Biogenic Amines

CAO Yuying, WANG Shuang, LIU Yining, GAO Manman, DU Liping, MA Lijuan, ZHANG Huiling, TIAN Xiaoju, YANG Weiming

(1. College of Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, China; 2. Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Key Laboratory of Tianjin Industrial Microbiology, Tianjin University of Science and Technology, Tianjin 300457, China; 3. School of Food and Wine, Ningxia University, Yinchuan 750021, China; 4. Ningxia Zhihuiyuan Stone Winery Co. Ltd., Yinchuan 750026, China)

Online:2023-06-25 Published:2023-06-30

摘要/Abstract

摘要： 为研究乳酸菌来源的漆酶酶学性质及其在生物胺降解中的应用潜力，以来源于植物乳杆菌的漆酶编码基因为研究对象，对其进行异源表达及酶学性质的研究，测定其最适温度和pH值、金属离子及螯合剂对漆酶活性的影响、重组漆酶对不同体积分数的乙醇的耐受性以及重组酶对生物胺的降解作用。结果表明，重组漆酶最适温度为40 ℃，最适pH值为4.0；低浓度的Mg2＋、Zn2＋和Ca2＋对重组漆酶活力具有不同程度的促进作用，其中Zn2＋和Ca2＋的促进作用最为明显；反之，高浓度的Mn2＋、Fe2＋和乙二胺四乙酸能够抑制重组漆酶活力至原本的15%甚至更低；2,2’-联氮双(3-乙基苯并噻唑啉-6-磺酸)为底物时重组酶的米氏常数为2.49 mmol/L，最大反应速率为0.22 mmol/（L·min）；重组漆酶对低体积分数乙醇具有很好的耐受性，在含有20%乙醇的体系中保存1 h后相对酶活力仍可达75%以上。生物胺降解实验表明，重组漆酶对酪胺和色胺的降解率达到了100%，对亚精胺、苯乙胺和组胺的降解率分别为49.60%、25.27%和21.80%。由此可见，本实验构建的来源于植物乳杆菌的重组漆酶针对低酒精度发酵饮品及发酵食品的生物胺降解具有很大的应用潜力。

关键词: 生物胺；漆酶；异源表达；酶学性质；植物乳杆菌

Abstract: In order to study the enzymatic properties of laccase derived from Lactobacillus and its potential application in biogenic amine degradation, the gene encoding laccase from Lactobacillus plantarum was heterologously expressed and the recombinant laccase was enzymatically characterized. The optimum reaction temperature and pH, the effects of metal ions and chelators on laccase activity, the tolerance of the recombinant laccase to different concentrations of ethanol and the degradation of biogenic amines by the recombinant laccase were determined. The results showed that the optimum reaction temperature and pH of the purified recombinant laccase was 40 ℃ and 4.0, respectively. Low concentrations of Mg2+, Zn2+ and Ca2+ could promote the activity of the recombinant laccase in different degrees, Zn2+ and Ca2+ being most effective. On the contrary, high concentrations of Mn2+, Fe2+ and ethylenediaminetetraacetic acid could inhibit the activity of recombinant laccase by 85% or more. Toward the substrate 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), the Michaelis constant and maximum reaction rate of the recombinant laccase were 2.49 mmol/L and 0.22 mmol/(L·min), respectively. In addition, the recombinant laccase had a good tolerance to ethanol at low concentration, and the residual activity was more than 75% after incubation in 20% (V/V) ethanol for 1 h. The degradation rates of tyramine and tryptamine by the recombinant laccase were both 100%, while the degradation rates of spermidine, phenethylamine and histamine were 49.60%, 25.27% and 21.80%, respectively. Based on the above results, the recombinant laccase from L. plantarum has great application potential in the degradation of biogenic amines in low-alcohol fermented beverages and fermented foods.

Key words: biogenic amines; laccase; heterologous expression; enzymatic properties; Lactobacillus plantarum

中图分类号:

Q939.99

曹玉莹，王爽，刘怡宁，高熳熳，杜丽平，马立娟，张惠玲，田晓菊，杨伟明. 植物乳杆菌漆酶的酶学性质及其对生物胺的降解作用[J]. 食品科学, 2023, 44(12): 157-163.

CAO Yuying, WANG Shuang, LIU Yining, GAO Manman, DU Liping, MA Lijuan, ZHANG Huiling, TIAN Xiaoju, YANG Weiming. Enzymatic Characteristics of Laccase from Lactobacillus plantarum and Its Effect on Degradation of Biogenic Amines[J]. FOOD SCIENCE, 2023, 44(12): 157-163.

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植物乳杆菌漆酶的酶学性质及其对生物胺的降解作用

Enzymatic Characteristics of Laccase from Lactobacillus plantarum and Its Effect on Degradation of Biogenic Amines

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