一种黑曲霉α-L-阿拉伯呋喃糖苷酶克隆表达、性质分析和果汁澄清效果

doi:10.7506/spkx1002-6630-20201105-052

食品科学 ›› 2022, Vol. 43 ›› Issue (2): 83-92.doi: 10.7506/spkx1002-6630-20201105-052

一种黑曲霉α-L-阿拉伯呋喃糖苷酶克隆表达、性质分析和果汁澄清效果

彭程，肖文熙，倪辉，李利君，谭万森，林燕玲，李清彪

（1.集美大学食品与生物工程学院，福建厦门 361021；2.集美大学港口与环境工程学院，福建厦门 361021；3.福建省食品微生物与酶工程重点实验室，厦门市食品生物工程技术研究中心，福建厦门 361021）

出版日期:2022-01-25 发布日期:2022-01-29
基金资助:
国家自然科学基金重点项目（U1805235）；厦门市科技局项目（3502Z20203058）

Cloning, Expression and Characterization of α-L-Arabinofuranosidase from Aspergillus niger and Its Clarification Effect on Fruit Juice

PENG Cheng, XIAO Wenxi, NI Hui, LI Lijun, TAN Wansen, LIN Yanling, LI Qingbiao

(1. College of Food and Biological Engineering, Jimei University, Xiamen 361021, China; 2. College of Harbour and Environmental Engineering, Jimei University, Xiamen 361021, China; 3. Key Laboratory of Food Microbiology and Enzyme Engineering Technology of Fujian Province, Research Center of Food Biotechnology of Xiamen City, Xiamen 361021, China)

Online:2022-01-25 Published:2022-01-29

摘要/Abstract

摘要： 利用基因重组技术克隆黑曲霉（Aspergillus niger）的1 887 bp α-L-阿拉伯呋喃糖苷酶基因，在毕赤酵母SMD1168中诱导表达获得α-L-阿拉伯呋喃糖苷酶。结果表明，重组蛋白大小为85 kDa，纯化后其比活力为55.73 U/mg；最适反应温度和pH值分别为65 ℃和5.0；金属离子K＋对该酶活力有促进作用，而Cu2＋对酶活力有明显抑制作用；以4-硝基苯酚-α-L-阿拉伯呋喃糖苷为底物时测得Km值和Vmax值分别为2.31 mmol/L和625 μmol/（mL·min）；该酶对柑橘果汁具有显著澄清效果。本研究不仅丰富了α-L-阿拉伯呋喃糖苷酶资源库，同时为后续探究α-L-阿拉伯呋喃糖苷酶在果汁加工中的应用提供了参考。

关键词: 黑曲霉；α-L-阿拉伯呋喃糖苷酶；异源表达；酶学性质；澄清

Abstract: The sequence of the 1 887 bp long α-L-arabinofuranosidase gene in Aspergillus niger was cloned by using gene recombination, and was expressed in Pichia pastoris SMD1168. The recombinant protein had a molecular mass of 85 kDa and a specific activity of 55.73 U/mg. The optimal temperature and pH for the enzyme were 65 ℃ and 5.0, respectively. The enzymatic activity was promoted by Mn2+, while it was significantly inhibited by K+. When p-nitrophenyl-α-L-arabinofuranoside (pNPA) was used as the substrate, the Km and Vmax values of α-L-arabinofuranosidase were 2.31 mmol/L and 625 μmol/(mL·min), respectively. More interestingly, the purified α-L-arabinofuranosidase showed a significant clarification effect on citrus juice. This study not only enriches the resource library of α-L-arabinofuranosidase, but also provides a reference for juice processing with α-L-arabinofuranosidase.

Key words: Aspergillus niger; α-L-arabinofuranosidase; heterologous expression; enzymatic properties; clarification

中图分类号:

彭程，肖文熙，倪辉，李利君，谭万森，林燕玲，李清彪. 一种黑曲霉α-L-阿拉伯呋喃糖苷酶克隆表达、性质分析和果汁澄清效果[J]. 食品科学, 2022, 43(2): 83-92.

PENG Cheng, XIAO Wenxi, NI Hui, LI Lijun, TAN Wansen, LIN Yanling, LI Qingbiao. Cloning, Expression and Characterization of α-L-Arabinofuranosidase from Aspergillus niger and Its Clarification Effect on Fruit Juice[J]. FOOD SCIENCE, 2022, 43(2): 83-92.

一种黑曲霉α-L-阿拉伯呋喃糖苷酶克隆表达、性质分析和果汁澄清效果

Cloning, Expression and Characterization of α-L-Arabinofuranosidase from Aspergillus niger and Its Clarification Effect on Fruit Juice

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