一株链霉菌的鉴定及其产格尔德霉素的发酵工艺研究  

Identification of a Streptomyces Strain and Study on the Fermentation Process of Geldanamycin Production

作　　者：杨鹭 袁源 方志锴[2] 林如[2] 江红[1,2] 周剑 YANG Lu;YUAN Yuan;FANG Zhi-kai;LIN Ru;JIANG Hong;ZHOU Jian(The School of Pharmacy,Fujian Medical University,Fuzhou 350122;Fujian Key Laboratory of Screening for Novel Microbial Products,Fujian Institute of Microbiology,Fuzhou 350007)

机构地区：[1]福建医科大学药学院,福州350122 [2]福建省微生物研究所福建省新药(微生物)筛选重点实验室,福州350007

出　　处：《生物技术通报》2024年第6期299-309,共11页Biotechnology Bulletin

基　　金：福建省社会发展引导性重点项目(2021Y0044);福建省属公益类科研院所基本科研专项(2023R1005004)。

摘　　要：【目的】对链霉菌Streptomyce sp.FIM18-0592进行菌种鉴定,并对其胞外格尔德霉素产量进行发酵工艺优化,旨在提高产量并降低发酵成本。【方法】通过形态特征、培养特征和生理生化特性,结合16S rDNA序列分析构建系统发育树进行菌种鉴定;采用单因素试验优化培养条件及培养基配方,进一步使用最陡爬坡试验和响应面试验优化培养基配方的含量。【结果】通过对链霉菌FIM18-0592的形态特征、培养特征及生理生化特征进行初步培养观察发现,其在ISP2等培养基上生长较好,气生菌丝旺盛,产黑色素。结合16S rDNA分子鉴定,确定该菌为格尔德霉素链霉菌(Streptomyces geldanamycininus)。通过单因素试验对发酵条件以及培养基配方进行优化,得到最适宜菌株发酵的培养条件为转速140 r/min、装液量12%(体积分数)、接种量7%(体积分数)、培养时间144 h。最佳的碳源、氮源、无机盐分别为葡萄糖、黄豆饼粉和硫酸铵。采用最陡爬坡试验和响应面优化试验确定了其最优的发酵培养基为:葡萄糖10.42%、黄豆饼粉1.68%、硫酸铵0.3%、乳酸0.3%、甘油4%、硫酸镁0.1%、碳酸钙0.4%,在此条件下,格尔德霉素的发酵效价达到2887μg/mL,较原始发酵工艺效价提高了66%。【结论】链霉菌FIM18-0592为格尔德霉素链霉菌,通过对其发酵工艺进行优化显著提高格尔德霉素的产量,为格尔德霉素及其衍生物的开发和利用奠定基础。【Objective】Streptomyces sp.FIM18-0592 was identified and its fermentation process was optimized for extracellular geldanamycin yield with the aim of increasing the yield and reducing the fermentation cost.【Method】Strain identification was carried out by morphological features,culture characteristics and physiological and biochemical properties,combined with 16S rDNA sequence analysis to construct a phylogenetic tree;a one-factor test was used to optimize the culture conditions and media formulations,and the contents of the media formulations were further optimized by the steepest-climbing test and the response surface test.【Result】Through preliminary culture observation of the morphological characteristics,culture characteristics and physiological and biochemical characteristics of Streptomyces geldanamycininus FIM18-0592,we found that it grew well on ISP2 and other media,with vigorous aerial mycelium and melanin production.Combined with 16S rDNA molecular identification,the Streptomyces sp.was identified as Streptomyces geldanamycininus.The fermentation conditions and medium formulation were optimized by one-factor test,and the most optimal culture conditions for the fermentation of the strain were speed of 140 r/min,liquid content of 12%,inoculum concentration of 7%(V/V),and fermentation time of 144 h.The optimal carbon source,nitrogen source,and mineral salt were glucose,soybean cake powder and NH4HPO4,respectively.The optimal fermentation medium was determined by steepest-climbing test and response surface optimization test as follows:Glucose 10.42%,soybean cake powder 1.68%,(NH4)2SO40.3%,lactic acid 0.3%,glycerol 4%,magnesium sulphate 0.1%,and calcium carbonate 0.4%.Under these conditions,the fermentation titer of geldanamycin reached 2887μg/mL,which was 66%higher than the original fermentation process.【Conclusion】Streptomyces sp.FIM18-0592 belongs to Streptomycete geldanamycininus,whose fermentation process is optimized to significantly increase the yield of geldanamycin,laying the foundati

关 键 词：格尔德霉素 菌种鉴定 发酵优化 单因素优化 响应面试验 形态特征 培养特性 

分 类 号：TQ927[轻工技术与工程—发酵工程]