植酸酶YiAPPA与生淀粉结合域SBD融合酶的构建及酶学性质分析

doi:10.13560/j.cnki.biotech.bull.1985.2017-0795

摘要/Abstract

摘要： 旨在获得酶学性质改良的植酸酶YiAPPA与生淀粉结合域SBD的融合酶。通过在植酸酶YiAPPA的C末端融合嗜热酸性α-淀粉酶GTamy的生淀粉结合域SBD，获得融合酶YiAPPA-SBD。酶学性质分析表明，YiAPPA-SBD的高温活性和热稳定性得到了提高，并获得了对生玉米淀粉的结合能力。其中YiAPPA-SBD于55-90℃范围内的相对酶活均高于YiAPPA的相对酶活；于80℃的半衰期提高约2倍；在生玉米淀粉浓度大于8%的条件下，YiAPPA-SBD对其结合率达到80%以上。并且YiAPPA-SBD保留有YiAPPA的其它优良酶学性质，最适反应pH为4.5，37℃的绝对酶活高达3 900 U/mg，具有优良的pH稳定性和蛋白酶抗性。

关键词: 植酸酶YiAPPA, 生淀粉结合域, 融合酶, 热稳定性, 生淀粉结合率

Abstract: This work aims to obtain a fusion enzyme with improved enzymatic properties comprised of phytase YiAPPA and raw-starch binding domain SBD. The raw-starch binding domain（SBD）of thermoacidophilic α-amylase GTamy was introduced into the C-terminal end of Yersinia intermedia phytase（YiAPPA）to generate a fusion enzyme YiAPPA-SBD. The fusion of YiAPPA with SBD resulted in improvements both in terms of thermal activity and stability. Besides，YiAPPA-SBD could absorb raw corn starch. Specifically，YiAPPA-SBD exhibited higher relative activity than YiAPPA within 55-90℃.The half-life of YiAPPA-SBD increased from 15 min to 30 min when incubated at 80℃. The adsorption efficiency of YiAPPA-SBD was 80% when the raw corn starch concentration was over 8%. Moreover，YiAPPA-SBD showed minor changes in its specific activity（3 900 U/mg）at optimal pH（pH 4.5），indicating that YiAPPA-SBD has solid pH stability and resistance to protease when compared to YiAPPA.

Key words: phytase YiAPPA, raw-starch binding domain, fusion enzyme, thermostability, raw-starch adsorption efficiency

袁林, 黄朝, 曾静, 郭建军, 张婷, 吕珺,. 植酸酶YiAPPA与生淀粉结合域SBD融合酶的构建及酶学性质分析[J]. 生物技术通报, 2018, 34(3): 200-207.

YUAN Lin, HUANG Zhao, ZENG Jing, GUO Jian-jun, ZHANG Ting, Lü Jun. Fusion of Phytase YiAPPA with the Raw-starch Binding Domain and Characterization of the Fusion Enzyme[J]. Biotechnology Bulletin, 2018, 34(3): 200-207.

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