食品与发酵工业 >

2019 , Vol. 45 >Issue 8: 22 - 28

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.019584

研究报告

来源于Wenyingzhuangia属海洋细菌的一种β-琼胶酶的克隆表达及性质研究

  • 田雪健 ,
  • 申晶晶 ,
  • 常耀光
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  • 1(中国海洋大学 食品科学与工程学院,山东 青岛,266003)
硕士研究生(常耀光副教授为通讯作者,E-mail:changyg@ouc.edu.cn)。

收稿日期: 2018-12-10

  修回日期: 2018-12-28

  网络出版日期: 2019-05-14

基金资助

国家自然科学基金(31671883);国家自然科学基金(U1606403)

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Cloning, expression, and characterization of a β-agarase from marine bacterium Wenyingzhuangia fucanilytica

  • TIAN Xuejian ,
  • SHEN Jingjing ,
  • CHANG Yaoguang
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  • 1(College of Food Science and Engineering, Ocean University of China, Qingdao 266003, China)

Received date: 2018-12-10

  Revised date: 2018-12-28

  Online published: 2019-05-14

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摘要

琼胶是一种重要的海洋多糖,其寡糖被证实具有丰富的生理调节功能。该研究拟从微生物基因组出发,利用生物信息学及分子生物学技术发掘新型琼胶酶。以海洋细菌Wenyingzhuangia fucanilytica CZ1127T为实验材料,从该菌的基因组中发掘出1条潜在的GH16家族β-琼胶酶编码基因序列aga16A,通过分子克隆、异源表达及分离纯化,证实其编码的蛋白Aga16A_Wf具有琼胶酶活力。酶学性质及作用方式分析表明,Aga16A_Wf的最适反应温度(50 ℃)高于琼胶的凝胶温度,且该酶展现出适冷性;Aga16A_Wf的动力学常数Km为3.6 g/L,Kcat为287.88 s-1;该酶为内切型水解酶,最小作用底物为四糖,最小产物为二糖。Aga16A_Wf可作为高效制备琼胶寡糖的工具酶,该酶的获取为琼胶及琼胶寡糖的进一步研究与应用提供了良好支撑。

关键词： β-琼胶酶; 克隆表达; 琼胶寡糖

本文引用格式

田雪健 , 申晶晶 , 常耀光 . 来源于Wenyingzhuangia属海洋细菌的一种β-琼胶酶的克隆表达及性质研究[J]. 食品与发酵工业, 2019 , 45(8) : 22 -28 . DOI: 10.13995/j.cnki.11-1802/ts.019584

Abstract

Agarose is an important marine polysaccharide and its oligosaccharides were considered to have a series of bioactive functions. A gene, aga16A, encoding a putative GH16 amily β-agarase was recovered from the genome of Wenyingzhuangia funcanilytica CZ1127T, and heterologously expressed and purified. The recombinant Aga16A_Wf was confirmed to be an agarase with the optimum reaction temperature of 50 ℃, higher than the gelling temperature of agar. Besides, the enzyme was active under cold environment. Its kinetic constants Km and Kcat were 3.6 g/L and 287.88 s-1, respectively. Aga16A_Wf was an endo-acting enzyme, its smallest substrate and product were tetrasaccharide and disaccharide, respectively. Aga16A_Wf could be served as a favorable tool to produce agar oligosaccharides efficiently. It is also useful for further studies on agar and its oligosaccharides.

Key words： β-agarase; cloning and expression; agar oligosaccharides

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