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几丁质和葡聚糖处理的异常威克汉姆酵母EG2中胞外几丁质酶和β-1,3-葡聚糖酶的抗真菌活性及表达模式

Antifungal activity and expression patterns of extracellular chitinase and β-1,3-glucanase in Wickerhamomyces anomalus EG2 treated with chitin and glucan.

作者信息

Hong Sin-Hyoung, Song Yong-Su, Seo Dong-Jun, Kim Kil-Yong, 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, South Korea.

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

出版信息

Microb Pathog. 2017 Sep;110:159-164. doi: 10.1016/j.micpath.2017.06.038. Epub 2017 Jun 28.

DOI:10.1016/j.micpath.2017.06.038
PMID:28668604
Abstract

In this study, the expression patterns of extracellular chitinase and β-1,3-glucanase from cultured Wickerhamomyces anomalus EG2 treated with chitin, glucan, and chemical chitinase inhibitors (kinetin, caffeine, and acetazolamide) were investigated using SDS-PAGE. Relationship between enzyme expression and antifungal activity from yeast plays a very important role for biocontrol of phytopathoges. To determine antifungal activity against phytopathogens, W. anomalus EG2 was shown to strongly inhibit hyphal growth of Fusarium oxysporum KACC 40032 and Rhizoctonia solani KACC 40111. Slight chitinase activity was observed 12 h after incubation in both PDB and YPD medium without colloidal chitin. The molecular weight of chitinase was approximately 124 kDa β-1,3-Glucanase isoenzyme (GN1 and GN2) was observed distinctly on SDS-PAGE gels when laminarin was used as a substrate. β-1,3-Glucanase isoenzyme was not observed when using glucan-containing high polymer complex (GHPC) as a substrate. Production of chitinase from W. anomalus EG2 was inhibited slightly by acetazolamide. Abnormal and cluster-shaped cells of W. anomalus EG2 were observed in both PDB and YPD medium treated with colloidal chitin. These results indicated that W. anomalus EG2 could be applied commercially as a biological control agent of phytopathogens and as a bioinhibitor of yeast cell growth.

摘要

在本研究中,使用SDS-PAGE研究了用几丁质、葡聚糖和化学几丁质酶抑制剂(激动素、咖啡因和乙酰唑胺)处理的培养异常威克汉姆酵母EG2的细胞外几丁质酶和β-1,3-葡聚糖酶的表达模式。酵母中酶表达与抗真菌活性之间的关系在植物病原菌的生物防治中起着非常重要的作用。为了确定对植物病原菌的抗真菌活性,异常威克汉姆酵母EG2被证明能强烈抑制尖孢镰刀菌KACC 40032和立枯丝核菌KACC 40111的菌丝生长。在不含胶体几丁质的PDB和YPD培养基中培养12小时后,观察到轻微的几丁质酶活性。几丁质酶的分子量约为124 kDa。当以海带多糖为底物时,在SDS-PAGE凝胶上可清楚地观察到β-1,3-葡聚糖酶同工酶(GN1和GN2)。当使用含葡聚糖的高聚物复合物(GHPC)作为底物时,未观察到β-1,3-葡聚糖酶同工酶。乙酰唑胺对异常威克汉姆酵母EG2几丁质酶的产生有轻微抑制作用。在用胶体几丁质处理的PDB和YPD培养基中均观察到异常威克汉姆酵母EG2的异常细胞和簇状细胞。这些结果表明,异常威克汉姆酵母EG2可作为植物病原菌的生物防治剂和酵母细胞生长的生物抑制剂进行商业应用。

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