产黄青霉形态代谢工程研究进展  

Metabolic engineering of Penicillium chrysogenum morphology

作　　者：郑之明[1] 赵根海[1] 王鹏[1] 刘会[1] 胡以华[2] 蔡晓春[2] 王丽[1] 贡国鸿[1] 桂芳[1] 王辉[1] 

机构地区：[1]中国科学院合肥物质科学研究院、中国科学院离子束生物工程学重点实验室,合肥230031 [2]中国人民解放军电子工程学院、脉冲功率激光技术国家重点实验室,合肥230037

出　　处：《科学通报》2014年第21期2017-2032,共16页Chinese Science Bulletin

基　　金：国家高技术研究发展计划(2009AA02Z305,2014AA021704)资助

摘　　要：丝状真菌广泛应用于工业微生物发酵,其发酵形态的多样性和调控复杂性为业界广泛关注.本文对产黄青霉的形态、调控以及与产量相关性展开研究.通过一种有效菌丝形态图像处理方法,获得菌丝形态特征参数,并应用于产黄青霉的形态动力学分析;以低能离子注入和低温空气等离子体对产黄青霉进行了复合诱变选育,比较产黄青霉形态突变株与原始菌的形态差异,揭示菌丝形态与次级代谢产物合成的相关性;以Ⅲ类几丁质合成酶基因chs4为目的基因,通过RNA干扰构建形态突变株,发现形态突变株菌丝变得粗短、分枝增多,转化株青霉素产量明显高于原始菌,其中PcRNAi1-17和PcRNAi2-1的青霉素产量分别比原始菌提高27%和41%,为产黄青霉形态代谢工程研究奠定了一定的理论基础.Filamentous fungi are widely used in the field of industrial fermentation, and their morphological diversity and complex regulation during fermentation have attracted serious attention from researchers. Penicillium chrysogenum morphology, intervention and regulation, as well as the relationship between morphology and yield, were researched in this study. We have established an effective image processing method to measure several hyphal morphological parameters. These parameters were applied to quantitative analysis of morphology and the metabolic status of the product under different culture conditions. Simultaneously, morphological dynamics equations were also established. Low energy ion beam implantation and low temperature air plasma techniques were used in this work to mutate P. chrysogenum. To reveal the relationship between hyphal morphology and the production of secondary metabolites, quantitative analysis of hyphal morphological variations in the fermentation process was also carried out with mutant strains and the original strain. We constructed an RNAi vector to silence the class III chitin synthase gene chs4 to construct morphological mutants. All transformants except PcRNAiI-15 exhibited enhanced penicillin yields, especially PcRNAiI-17 and PcRNAi2-1, whose yields were 27% and 41% higher than the original strain. Our results provide a theoretical basis for understanding the regulatory mechanisms of penicillin production.

关 键 词：图像处理 诱变 形态突变株 青霉素产量 几丁质合成酶 RNA干扰 

分 类 号：TQ927[轻工技术与工程—发酵工程]