生物技术通报 ›› 2021, Vol. 37 ›› Issue (3): 44-52.doi: 10.13560/j.cnki.biotech.bull.1985.2020-0800

• 研究报告 • 上一篇    下一篇

黑曲霉单宁酶基因Tan2克隆与表达

李红叶1(), 陈立佼1, 刘明丽1,3, 郭天杰1,3, 王道平2, 潘映红2, 赵明1,3()   

  1. 1.云南农业大学龙润普洱茶学院,昆明 650201
    2.中国农业科学院作物科学研究所/ 国家农作物基因资源与基因改良重大科学工程,北京 100081
    3.云南农业大学云南省药用植物生物学重点实验室 云南农业大学西南中药材种质创新与利用国家地方联合工程研究中心,昆明 650201
  • 收稿日期:2020-06-30 出版日期:2021-03-26 发布日期:2021-04-02
  • 作者简介:李红叶,女,硕士研究生,研究方向:茶树病害;E-mail:837356337@qq.com
  • 基金资助:
    国家自然科学基金项目(31760225);云南省中青年学术技术带头人后备人才培养项目(2017HB026)

Cloning and Expression of Tannase Gene Tan2 from Aspergillus niger

LI Hong-ye1(), CHEN Li-jiao1, LIU Ming-li1,3, GUO Tian-jie1,3, WANG Dao-ping2, PAN Ying-hong2, ZHAO Ming1,3()   

  1. 1. College of Longrun Pu-erh Tea,Yunnan Agricultural University,Kunming 650201
    2. Institute of Crop Sciences,Chinese Academy of Agricultural Sciences/National Key Facility for Crop Gene Resources and Genetic Improvement,Beijing 100081
    3. The Key Laboratory of Medicinal Plant Biology of Yunnan Province/National & Local Joint Engineering Research Center on Germplasm Innovation & Utilization of Chinese Medicinal Materials in Southwestern China,Yunnan Agricultural University,Kunming 650201
  • Received:2020-06-30 Published:2021-03-26 Online:2021-04-02

摘要:

单宁酶(Tannase,EC 3.1.1.20)能水解单宁中的酯键和羧酚酸键,产生没食子酸以及对应醇,在食品、饮料、饲料、制药、医药、化妆品等各类工业中应用广泛,也在普洱茶发酵中具有重要作用。从普洱茶发酵中分离的黑曲霉菌株PU001中克隆得到单宁酶基因Tan2,并连接到表达载体pCold-Ⅰ构建BL21-pCold Ⅰ原核冷诱导表达系统,转化至感受态大肠杆菌BL21中,SDS-PAGE与质谱鉴定均表明单宁酶Tan 2表达成功,旨在为进一步研究该酶在普洱茶发酵中的作用机制以及其他领域的应用奠定基础。

关键词: 单宁酶, 黑曲霉, 普洱茶

Abstract:

Tannase(EC 3.1.1.20)can hydrolyze the ester bond and carboxyphenolic acid bond in tannins to produce gallic acid and corresponding alcohols. It is widely used in food,beverage,feed,pharmacy,medicine,cosmetics and other industries,and also plays an important role in the fermentation of Pu-erh tea. In this work,tannase gene Tan 2 was cloned from Aspergillus niger PU001,which was previously isolated from fermentation in Pu-erh tea. It was ligated with expression vector pCold-I to construct the prokaryotic cold inducible expression system of BL21-pCold I,which was transformed into Escherichia coli BL21. SDS-PAGE and mass spectrometry identification showed that tannase Tan 2 was expressed successfully. This result provides a basis for further survey on the role of tannase in the fermentation of Pu-erh tea and application in other fields.

Key words: tannase, Aspergillus niger, Pu-erh tea