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首页> 外文期刊>Applied Microbiology and Biotechnology >Morphological changes induced by class III chitin synthase gene silencing could enhance penicillin production of Penicillium chrysogenum
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Morphological changes induced by class III chitin synthase gene silencing could enhance penicillin production of Penicillium chrysogenum

机译：III类几丁质合酶基因沉默诱导的形态变化可增强产黄青霉的青霉素产量

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Chitin synthases catalyze the formation of β-(1,4)-glycosidic bonds between N-acetylglucosamine residues to form the unbranched polysaccharide chitin, which is the major component of cell walls in most filamentous fungi. Several studies have shown that chitin synthases are structurally and functionally divergent and play crucial roles in the growth and morphogenesis of the genus Aspergillus although little research on this topic has been done in Penicillium chrysogenum. We used BLAST to find the genes encoding chitin synthases in P. chrysogenum related to chitin synthase genes in Aspergillus nidulans. Three homologous sequences coding for a class III chitin synthase CHS4 and two hypothetical proteins in P. chrysogenum were found. The gene which product showed the highest identity and encoded the class III chitin synthase CHS4 was studied in detail. To investigate the role of CHS4 in P. chrysogenum morphogenesis, we developed an RNA interference system to silence the class III chitin synthase gene chs4. After transformation, mutants exhibited a slow growth rate and shorter and more branched hyphae, which were distinct from those of the original strain. The results also showed that the conidiation efficiency of all transformants was reduced sharply and indicated that chs4 is essential in conidia development. The morphologies of all transformants and the original strain in penicillin production were investigated by light microscopy, which showed that changes in chs4 expression led to a completely different morphology during fermentation and eventually caused distinct penicillin yields, especially in the transformants PcRNAi1-17 and PcRNAi2-1 where penicillin production rose by 27 % and 41 %, respectively.
机译：几丁质合酶催化N-乙酰氨基葡萄糖残基之间形成β-（1,4）-糖苷键，形成无支链的多糖几丁质，这是大多数丝状真菌细胞壁的主要成分。几项研究表明，几丁质合酶在结构和功能上存在差异，并且在曲霉属的生长和形态发生中起着关键作用，尽管在黄青霉中对此主题的研究很少。我们使用BLAST在产黄青霉中发现与构巢曲霉中的几丁质合酶基因相关的编码几丁质合酶的基因。在产黄青霉中发现了三个编码Ⅲ类几丁质合酶CHS4的同源序列和两个假想蛋白。详细研究了该产物显示出最高的同一性并编码III类几丁质合酶CHS4的基因。为了研究CHS4在产黄青霉的形态发生中的作用，我们开发了一种RNA干扰系统来沉默III类几丁质合酶基因chs4。转化后，突变体的生长速度较慢，菌丝较短且分支较多，这与原始菌株不同。结果还表明，所有转化体的分生孢子效率都急剧降低，表明chs4在分生孢子的形成中至关重要。通过光学显微镜研究了所有转化子的形态和青霉素生产中的原始菌株，结果表明chs4表达的变化导致了发酵过程中形态的完全不同，并最终导致了不同的青霉素产量，尤其是在转化子PcRNAi1-17和PcRNAi2-中。 1青霉素产量分别增长27％和41％。

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