高产核糖核酸马克斯克鲁维酵母的选育及发酵培养基优化  

Breeding and fermentation medium optimization of Kluyveromyces marxianus with high-yield ribonucleic acid

作　　者：郭灿 卢东霞 张鹏杰 陈叶福 肖冬光 郭学武 GUO Can;LU Dongxia;ZHANG Pengjie;CHEN Yefu;XIAO Dongguang;GUO Xuewu(Key Laboratory of Industrial Fermentation Microbiology,Ministry of Education,Tianjin Industrial Microbiology Key Laboratory,College of Biotechnology,Tianjin University of Science and Technology,Tianjin 300457,China)

机构地区：[1]天津科技大学,生物工程学院,工业发酵微生物教育部重点实验室,天津市工业微生物重点实验室,天津300457

出　　处：《中国酿造》2024年第12期88-95,共8页China Brewing

基　　金：黔科合平台人才(CXTD[2023]029)。

摘　　要：核糖核酸(RNA)是生物体的重要组成部分,被广泛应用于食品和制药工业。该研究以可利用乳糖的马克斯克鲁维酵母(Kluyveromyces marxianus)TY-20为亲本菌株,采用同源重组技术过表达己糖转运蛋白编码基因HXT14、HXT15,构建重组菌株TY20-HXT14、TY20-HXT15,以期提高菌株的乳糖利用率与RNA含量。在此基础上,以胞内RNA含量为评价指标,采用单因素试验及响应面试验对重组菌株TY20-HXT14的培养基组成进行优化。结果表明,重组菌株TY20-HXT14的生长性能较好、胞内RNA含量最高,其在乳糖培养基中培养至对数期(OD_(600 nm)=1.000)时,胞内RNA含量较亲本菌株TY-20提高14.48%。重组菌株TY20-HXT14产RNA的最佳培养基组成:乳糖添加量14.8%,酵母浸粉添加量1.04%,磷酸二氢钾添加量为0.31%,硫酸镁添加量0.2%。在此培养基组成下,重组菌株TY20-HXT14的胞内RNA含量达到18.04%,与优化前相比(14.23%)提高了26.77%。Ribonucleic acid(RNA)is an important building block of living organisms and widely used in the food and pharmaceutical industries.In this study,recombinant strains TY20-HXT14 and TY20-HXT15 were constructed using lactose-capable Kluyveromyces marxianus TY-20 as the parent strain,and hexose transporter coding genes HXT14 and HXT15 were overexpressed by homologous recombination technology,in order to improve the lactose utilization and RNA content of the strain.On this basis,using the intracellular RNA content as evaluation index,the medium composition of the recombinant strain TY20-HXT14 was optimized by single factor tests and response surface tests.The results showed that the recombinant strain TY20-HXT14 had optimal growth performance and the highest intracellular RNA content.When cultured in lactose medium to the logarithmic stage(OD_(600 nm)=1.000),the intracellular RNA content of TY20-HXT14 was 14.48%higher than that of the parental strain TY-20.The optimal medium composition for RNA production of the recombinant strain TY20-HXT14 was lactose addition 14.8%,yeast extract powder addition 1.04%,KH2PO_(4) addition 0.31%and MgSO_(4) addition 0.2%.Under the optimal medium composition,the intracellular RNA content of the recombinant strain TY20-HXT14 reached 18.04%,which was 26.77%higher than that before optimization(14.23%).

关 键 词：核糖核酸 马克斯克鲁维酵母 同源重组 培养基优化 乳糖 

分 类 号：TQ926[轻工技术与工程—发酵工程]