在吸水链霉菌中表达透明颤菌血红蛋白基因提高子囊霉素产量  

Enhancement of Ascomycin Production by Expression of Vitreoscilla Hemoglobin Gene in Streptomyces hygroscopicus

作　　者：余志拓 沈晓放[1] 吴远杰[1] 杨松柏[1] 陈少欣[1] YU Zhituo SHEN Xiaofang WU Yuanjie YANG Songbai CHEN Shaoxin(State Key Lab. of New Drug and Pharmaceutical Process, Shanghai Institute of Pharmaceutical Industry, China State Institute of Pharmaceutical Industry, Shanghai 201203)

机构地区：[1]中国医药工业研究总院上海医药工业研究院创新药物与制药工艺国家重点实验室,上海201203

出　　处：《中国医药工业杂志》2017年第6期844-849,共6页Chinese Journal of Pharmaceuticals

基　　金：国家"重大新药创制"科技重大专项(2014ZX09201001-005-001)

摘　　要：为了解决子囊霉素(1)发酵过程低溶氧的问题,本研究将透明颤菌(Vitreoscilla)血红蛋白基因(vgb)整合至1生产菌株吸水链霉菌(Streptomyces hygroscopicus)中,构建工程菌SIPI-FK520-2,通过RT-PCR分析检测到了vgb基因的转录表达,进一步利用CO结合差光谱分析也证实了工程菌中透明颤菌血红蛋白(VHb)具有生物活性。最后在发酵罐进行该工程菌的发酵培养研究,结果表明,与野生菌相比,工程菌发酵过程溶氧水平明显改善,1产量提高了38%,达到1 269.7 mg/L。In order to solve the shortage of dissolved oxygen in the fermentation process of ascomycin （1）,the Vitreoscilla hemoglobin gene （vgb） which encodes Vitreoscilla hemoglobin （VHb） was integrated into 1 producing strain Streptomyces hygroscopicus. The codon-optimized vgb gene was ligased with plasmid pSET-M, making it under the control of constitutive erythromycin resistance gene promoter （ermE＊）, to yield an recombinant plasmid pSET-vgb.Then, vgb gene was introduced into the genome of S. hygroscopicus SIPI-FK520-1 through conjugal transfer to construct an engineering strain named SIPI-FK520-2. The RT-PCR analysis showed that vgb gene had been successfully expressed in the recombinant engineering strain. The biological activity of VHb in the recombinant strain was further detected and confirmed by CO-binding difference spectroscopy. Finally, the strain was cultured in a 5 L bioreactor. The results indicated that the level of dissolved oxygen in fermentation process was significantly improved, and the yield of 1 was increased by 38% compared to the wild strain, up to 1 269.7 mg/L.

关 键 词：子囊霉素 透明颤菌血红蛋白 吸水链霉菌 基因工程 溶氧 大环内酯抗生素 抗菌药 免疫抑制药 

分 类 号：TQ469[化学工程—制药化工]