龟裂链霉菌zwf2基因敲入及阻断对土霉素合成影响.docVIP

龟裂链霉菌zwf2基因敲入及阻断对土霉素合成影响摘要：目的考察敲入及阻断zwf2基因对6-磷酸葡糖脱氢酶（G6PDH）活性、细胞生长、土霉素合成的影响。方法构建敲入转化子M4018-(zwf2)和阻断转化子M4018-Δzwf2后，以原始菌株为对照，比较敲入转化子菌株M4018-(zwf2)和阻断转化子菌株M4018-Δzwf2摇瓶发酵后G6PDH活性、细胞生长、土霉素合成等生物性质的变化。结果阻断转化子菌株M4018-Δzwf2的G6PDH活性是原始菌的50 %左右，敲入转化子菌株M4018-(zwf2) 的G6PDH活性则略高于原始菌；阻断转化子菌株M4018-Δzwf2土霉素生物合成水平比原始菌提高27 %，比敲入转化子菌株M4018-(zwf2)高约40 %；发酵结束时敲入转化子菌株M4018-(zwf2)的细胞干重高于其他2株菌，其它2株菌的细胞干重差异不大。结论zwf2阻断有利于土霉素的生物合成，zwf2增强则对细胞生长有利。 关键词：6-磷酸葡糖脱氢酶；zwf2；龟裂链霉菌M4018；土霉素生物合成 中图分类号：Q939.97文献标识码：A文章编号：1672-979X(2009)01-0007-04 Effects of zwf2 Gene Knock-in and Disruption on Oxytetracycline Biosynthesis in Streptomyces rimosus M4018 LIAO Yu-ling 1, LIU Zhi-yong2, TANG Zhen-yu2, GUO Mei-jin1,2*, ZHUANG Ying-ping2, ZHANG Si-liang2 (1. Department of Bioengineering, Jiangxi Agricultural University, Nanchang 330045, China; 2. State Key Laboratory of Bioreactor Engineering, East China University of Science Technology, Shanghai 200237,China) Abstract：Objective To invesgate the effects of zwf2 gene knock-in and disruption on the specific activity of glucose-6-phosphate dehydrogenase (G6PDH), cell growth and oxytetracycline (OTC) biosynthesis in Streptomyces rimosus M4018. Methods Taking parent strain S. rimosus M4018 as control, the changes of the specific acitivity of G6PDH, cell growth and OTC level in the media were measured after the mutants M4018-Δzwf2 and M4018-(zwf2) respectively obtained through gene disruption and knock-in were cultured in shaking flask. Results The specific acitivity of G6PDH in mutant M4018-Δzwf2 was only half of that in parent strain, while the specific acitivity of G6PDH in mutant M4018-(zwf2) was a bit higher than that in parent strain over the fermentation period. However, the OTC level in mutant M4018-Δzwf2was 27 % and 40 % higher than that in parent strain and mutant M4018-(zwf2), respectively. As for the cell growth, the dry cell weight of mutant M4018-(zwf2) was the highest and the difference between mutant M4018-Δzwf2 and parent strain w