Development of Intergeneric Conjugal Gene Transfer System in Streptomyces diastatochromogenes 1628 and Its Application for Improvement of Toyocamycin Production
Development of Intergeneric Conjugal Gene Transfer System in Streptomyces diastatochromogenes 1628 and Its Application for Improvement of Toyocamycin Production
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DOI：
10.1007/s00284-013-0461-z
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发表时间：
2014-02-01
影响因子：
2.6
通讯作者：
Yu, Xiaoping
中科院分区：
文献类型：
--
作者：
Ma, Zheng;Liu, Jinxiu;Yu, Xiaoping
异变色链霉菌（Streptomyces diastatochromogenes） 1628是一种能产生丰霉素（toyocamycin， TM）的菌株，在抑制农业病原菌的发展方面具有潜在的生物防治作用。在本研究中，利用属间偶联建立了一个高效的转化体系。这是通过优化实验参数实现的。在最佳条件下，每个受体的最大共轭频率为4.1 x 10（-4）。为了在S. diastatochromogenes 1628中异源表达编码玻璃体颤菌血红蛋白的基因vgb，我们将vgb置于组成启动子PermE（*）的控制下，构建质粒pIB139-vgb。利用上述建立的属间偶联将该质粒整合到S. diastatochromogenes 1628的染色体上。最终筛选出TM产量最高的菌株1628-VHB-23。结果表明，在不同的溶解氧条件下，vgb基因的表达均能显著促进S. disastatochromogenes 1628的细胞生长和TM的产生。特别是在有限好氧条件下，菌株1628-VHB-23在7-l发酵罐中的DCW和TM分别比野生菌株高33.3%和210%。
Streptomyces diastatochromogenes 1628, capable of producing toyocamycin (TM), has exhibited a potential biocontrol effect in inhibiting the development of phytopathogens in the agriculture field. In this study, an efficient transformation system was developed using the intergeneric conjugation. This was achieved by optimization of experimental parameters. Under optimal conditions, a maximal conjugation frequency of 4.1 x 10(-4) per recipient was obtained. In order to heterologously express the gene vgb encoding Vitreoscilla hemoglobin in S. diastatochromogenes 1628, we placed vgb under the control of the constitutive promoter PermE(*) and constructed plasmid pIB139-vgb. This plasmid was integrated into the chromosome of S. diastatochromogenes 1628 using intergeneric conjugation established above. Finally, strain 1628-VHB-23 with the highest TM production was screened. Results indicated that expression of vgb gene had always significantly promoted the cell growth and TM production in S. diastatochromogenes 1628 under different dissolved oxygen conditions. In particular, under the limited aerobic condition, strain 1628-VHB-23 obtained 33.3 % more DCW and produced 210 % more TM in 7-l fermentor as compared with the wild-type strain.