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耐冷性Pedobacter sp. PR-M6几丁质酶的鉴定、纯化、表达模式及体外抗真菌活性

Identification, purification, and expression patterns of chitinase from psychrotolerant Pedobacter sp. PR-M6 and antifungal activity in vitro.

作者信息

Song Yong-Su, Seo Dong-Jun, Jung Woo-Jin

机构信息

Department of Agricultural Chemistry, Institute of Environmentally-Friendly Agriculture (IEFA), College of Agricultural and Life Science, Chonnam National University, Gwangju 61186, Republic of Korea.

Department of Agricultural Chemistry, Institute of Environmentally-Friendly Agriculture (IEFA), College of Agricultural and Life Science, Chonnam National University, Gwangju 61186, Republic of Korea.

出版信息

Microb Pathog. 2017 Jun;107:62-68. doi: 10.1016/j.micpath.2017.03.018. Epub 2017 Mar 19.

Abstract

In this study, a novel psychrotolerant chitinolytic bacterium Pedobacter sp. PR-M6 that displayed strong chitinolytic activity on 0.5% colloidal chitin was isolated from the soil of a decayed mushroom. Chitinase activity of PR-M6 at 25 °C (C25) after 6 days of incubation with colloidal chitin increased rapidly to a maximum level (31.3 U/mg proteins). Three chitinase isozymes (chiII, chiIII, and chiIV) from the crude enzyme at 25 °C (C25) incubation were expressed on SDS-PAGE gels at 25 °C. After purification by chitin-affinity chromatography, six chitinase isozymes (chiI, chiII, chiIII, chiIV, chiV, and chiVI) from C25-fractions were expressed on SDS-PAGE gels at 25 °C. Major bands of chitinase isozymes (chiI, chiII, and chiIII) from C4-fractions were strongly expressed on SDS-PAGE gels at 25 °C. Pedobacter sp. PR-M6 showed high inhibition rate of 60.9% and 57.5% against Rhizoctonia solani and Botrytis cinerea, respectively. These results indicated that psychrotolerant Pedobacter sp. PR-M6 could be applied widely as a microorganism agent for the biocontrol of agricultural phytopathogens at low temperatures.

摘要

在本研究中,从腐烂蘑菇的土壤中分离出一种新型耐冷几丁质分解细菌食杆菌属菌株PR-M6,该菌株在0.5%的胶体几丁质上表现出较强的几丁质分解活性。与胶体几丁质在25℃(C25)孵育6天后,PR-M6的几丁质酶活性迅速增加至最高水平(31.3 U/mg蛋白质)。25℃孵育的粗酶中的三种几丁质酶同工酶(chiII、chiIII和chiIV)在25℃的SDS-PAGE凝胶上表达。通过几丁质亲和层析纯化后,来自C25组分的六种几丁质酶同工酶(chiI、chiII、chiIII、chiIV、chiV和chiVI)在25℃的SDS-PAGE凝胶上表达。来自C4组分的几丁质酶同工酶(chiI、chiII和chiIII)的主要条带在25℃的SDS-PAGE凝胶上强烈表达。食杆菌属菌株PR-M6对立枯丝核菌和灰葡萄孢的抑制率分别高达60.9%和57.5%。这些结果表明,耐冷食杆菌属菌株PR-M6作为低温下农业植物病原菌生物防治的微生物制剂具有广泛的应用前景。

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