食品与发酵工业 >

2021 , Vol. 47 >Issue 13: 72 - 78

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.026774

研究报告

降解黄曲霉毒素B1的乳酸片球菌重组多铜氧化酶学性质

  • 刘畅 ,
  • 马现永 ,
  • 马三梅 ,
  • 邓盾
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  • 1(暨南大学 生物工程学系,广东 广州,510632)
    2(广东省农业科学院动物科学研究所 畜禽育种国家重点实验室 农业农村部华南动物营养与饲料重点实验室 广东省畜禽育种与营养研究重点实验室 广东畜禽肉品质量安全控制与评定工程技术研究中心,广东 广州,510640)
硕士研究生(邓盾助理研究员和马三梅副研究员为共同通讯作者,E-mail:dengdun2008@126.com,wyfmsm@163.com)

收稿日期: 2021-01-20

  修回日期: 2021-03-03

  网络出版日期: 2021-08-02

基金资助

国家自然科学基金项目(C31802103);广东省自然科学基金项目(2018A030313004);广州市科技计划项目(2018040100224);清远市创新创业科研团队项目(2018002);广东省现代农业产业技术体系创新团队项目(2019KJ112,2019KJ115);广州市民生科技计划项目(201903010083)

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Enzymatic properties of aflatoxin B1 degradation by recombinant copper oxidase from Pediococcus acidilactici

  • LIU Chang ,
  • MA Xianyong ,
  • MA Sanmei ,
  • DENG Dun
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  • 1(Department of bioengineering,Jinan University,Guangzhou 510632,China)
    2(Institute of Animal Science,Guangdong Academy of Agricultural Sciences;State Key Laboratory of Livestock and Poultry Breeding;Key Laboratory of Animal Nutrition and Feed Science in South China,Ministry of Agriculture;Guangdong Key Laboratory of Animal Breeding and Nutrition;Guangdong Engineering Technology Research Center of Animal Meat quality and Safety Control and Evaluation,Guangzhou 510640,China)

Received date: 2021-01-20

  Revised date: 2021-03-03

  Online published: 2021-08-02

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摘要

主要对1株乳酸片球菌Pediococcus acidilactici 7_4中的多酮氧化酶 (multicopper oxidase,MCO) 基因CueO进行重组表达和降解黄曲霉毒素B1(Alfatoxin B1,AFB1)的能力测定。通过基因工程的手段将CueO基因克隆连接到pET-28a(+)中,并转化到大肠杆菌 (Escherichia coli) 中进行异源表达,得到可溶性表达的重组蛋白。并进一步研究了温度、pH和底物浓度等因素对重组蛋白降解AFB1的影响以及降解产物的细胞毒性。结果表明,重组多铜氧化酶降解AFB1的最佳介体为丁香醛 (syringaldehyde,SA),最适pH为7.0,最适温度为37 ℃,该酶Km=1.99×10-3 μmol,Vmax=11.31 pmol/(min·mg),重组CueO对AFB1的降解产物对肝细胞的毒性明显降低。

关键词： 乳酸片球菌; 多酮氧化酶; 黄曲霉毒素B1; 漆酶; 酶学性质

本文引用格式

刘畅 , 马现永 , 马三梅 , 邓盾 . 降解黄曲霉毒素B1的乳酸片球菌重组多铜氧化酶学性质[J]. 食品与发酵工业, 2021 , 47(13) : 72 -78 . DOI: 10.13995/j.cnki.11-1802/ts.026774

Abstract

To detoxify AFB1 was a concern in the feed and food industry. In this paper, a multicopper oxidase gene from Pediococcus acidilactici 7_4 was heterogeneously expressed and the ability to degrade AFB1 were determined. The gene of CueO was cloned into pET-28a(+) by genetic engineering methods and could be soluble expressed in Escherichia coli. The effects of temperature, pH and substrate concentration on AFB1 degradation rate were studied. The best mediator for recombinant laccase was syringaldehyde (SA). The recombinant laccase displayed the highest degradation rate for AFB1 at 37°C in phosphate buffer (pH 7.0). The Km and Vmax value were 1.99×10-3 μmol and 11.31 pmol/(min·mg), respectively. The degradation products of AFB1 by recombinant expressed CueO were significantly less toxic to liver cells.

Key words： Pediococcus acidilactici; multicopper oxidase; aflatoxin B1; laccase; enzymatic properties

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