Position: Home > Articles > Effects of Overexpressing Hemoprotein Gene from Sinorhizobium meliloti on Nosiheptide Yield of Streptomyces actuosus
Journal of Anhui Agricultural Sciences
2016,44
(24)
157-159+162
过表达根瘤血红蛋白基因对活跃链霉菌那西肽产量的影响
作 者：
莫秋燕;宋元达
单 位：
江南大学食品学院食品与健康研究中心
关键词：
根瘤菌血红蛋白;活跃链霉菌;那西肽
摘 要：
[目的]采用过表达根瘤血红蛋白基因改善活跃链霉菌的溶氧表达,以提高那西肽的产量。[方法]通过全基因组合成的方法得到根瘤菌血红蛋白基因(sm),连接到整合型质粒p IB139上,构建表达载体p IB139-sm。通过大肠杆菌-链霉菌结合转移的方法将sm基因整合到活跃链霉菌基因组上,通过PCR验证各转化子目的片段已整合到活跃链霉菌基因组上,并进行大量筛选得到重组菌株。[结果]经SDS-PAGE和CO结合差示光谱分析,结果显示,目的蛋白均已成功表达且具有相应的蛋白活性。摇瓶培养结果显示,Smf Hb表达对菌体生长和那西肽产量有明显的促进作用,过表达Smf Hb的重组菌株那西肽产量达1 245.7μg/m L,比原始菌株提高39.6%。[结论]过表达sm基因提高那西肽产量的作用优于vhb。
译 名：
Effects of Overexpressing Hemoprotein Gene from Sinorhizobium meliloti on Nosiheptide Yield of Streptomyces actuosus
作 者：
MO Qiu-yan;SONG Yuan-da;Food and Health Research Center,School of Food and Health,Jiangnan University;
关键词：
Sinorhizobium meliloti;;Streptomyces actuosus;;Nosiheptide
摘 要：
[Objective] To improve the dissolved oxygen expression of Streptomyces actuosus by overexpressing hemoprotein Gene from Sinorhizobium meliloti,and to enhance the yield of nosiheptide. [Method] Sinorhizobium meliloti hemoglobin gene( sm) was obtained through the whole gene synthesis method,which were cloned into the plasmid p IB139 to construct expression vectors p IB139-sm. The plasmid containing sm was introduced into Streptomyces actuosus through conjugal transfer between E. coli and Streptomces. The transformants were verified by PCR,and then were purified by several cycles of single-colony screen. [Result]The expression and activity of Smf Hb was confirmed by SDSPAGE and CO-difference spectrum analysis,respectively. Flask culture experiment results showed that the nosiheptide yield in Smf Hb-overexpressing strain( 1 245. 7 μg / m L) was increased by 39. 6%( P < 0. 05) compared with the control strain( 892. 5 μg / m L). [Conclusion]Overexpressing sm gene is superior to vhb in enhancing the yield of nosiheptide yield.
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