Armillariella tabescens EJLY2098β-甘露聚糖酶的诱导、纯化及酶学性质分析

中国生物工程杂志

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Armillariella tabescens EJLY2098β-甘露聚糖酶的诱导、纯化及酶学性质分析

姚冬生黄小葵刘大岭谢春芳胡熔

暨南大学生命科学院生物技术实验室暨南大学生命科学院生物技术实验室暨南大学生命科学院生物技术实验室暨南大学生命科学院生物技术实验室暨南大学生命科学院生物技术实验室

Inducement, Purification and Characterization of β-mannanase from Armillariella tabescens EJLY2098

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摘要： Armillariella tabescens EJLY2098经魔芋精粉诱导，可产β-甘露聚糖酶，再用正交实验优化诱导培养基，结果在培养基为魔芋精粉2％、蛋白胨1％、土豆汁25％、KH2PO4 0.3％、MgSO4·7H2O 0.15％、维生素B1 0.01％时可诱导出较高活性的酶。用DEAE－阴离子交换色谱从培养上清分离纯化β-甘露聚糖酶，出现2个活性组份。活性组份P2 的SDS-PAGE分析，发现为电泳纯的一条带，分子量约78.9kD。 HPTLC分析， P2为内切β-甘露聚糖酶；酶反应的最适温度为60℃，最适pH 为4.0～6.0。保温30min的半失活温度t1/2为63℃，在pH 4.5～6.0之间稳定性较好。Na+和Ba2+对其有激活作用，等电点pI约为4.0～4.1。本研究获得了一株产β-甘露聚糖酶的新菌种，为进一步用基因工程方法克隆并构建具有完整自主知识产权的重组β-甘露聚糖酶基因工程菌提供了一个重要的基础工作。

Abstract: Armillariella tabescens EJLY2098 was capable of secreting β-mannanase by induced with konjac. A 34 orthogonal design was applied to determine the optimum medium of inducing mannanase by Armillariella tabescens EJLY2098. The results suggested that Armillariella tabescens EJLY2098 secreted the high activity enzyme by the optimum medium, which composed of 2% konjac, 1% peptone,25% potato juice.，0.3% KH2PO4,15% MgSO4·7H2O, 0.01％ VitB1。Purified by DEAE-anion exchange chromatography, two eluting peaks(P1 and P2)had activity of β-mannanase were obtained, and one of them(named β-mannanase P2) was a single band on the SDS-PAGE, and the molecular weight of β-mannanase P2 was 78.9kD.The isoelectric point of β-mannanase P2 was estimated to be 4.0~4.1. The optimum activity for the enzyme was found at 60℃ and pH4.0～6.0, and the enzyme was stable between pH4.5~6.0. The activity ofβ-mannanase P2 were enhanced by Na+ and Ba2+.Thisβ-mannanase can be used in feed industy and in this works, the authors obtained a new fungi secreting β-mannanase, the work improved an important base for cloning mannanase gene and constructing recombined microbe expressed β-mannanase .

收稿日期: 2006-01-27 出版日期: 2006-07-25

通讯作者: 刘大岭

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	黄小葵
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引用本文:

姚冬生,黄小葵,刘大岭,谢春芳,胡熔. Armillariella tabescens EJLY2098β-甘露聚糖酶的诱导、纯化及酶学性质分析[J]. 中国生物工程杂志, .

. Inducement, Purification and Characterization of β-mannanase from Armillariella tabescens EJLY2098. China Biotechnology, .

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https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2006/V26/I07/0

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