氧化葡萄糖酸杆菌WSH-003中D-山梨醇脱氢酶关键基因的表达分析  

Analysis of D-Sorbitol Dehydrogenase Gene in Gluconobacter oxydans WSH-003

作　　者：胡于东 堵国成[1,2] 陈坚[1,2] 周景文[1,2] HU Yudong DU Gocheng CHEN Jia ZHOU Jingwen(Key Laboratory of Industrial Biotechnology, Ministry of Education, ,liangnan University, Wuxi 214122, China School of Biotcchnology, Jiangnan University, Wuxi 214122, China)

机构地区：[1]江南大学工业生物技术教育部重点实验室,江苏无锡214122 [2]江南大学生物工程学院,江苏无锡214122

出　　处：《食品与生物技术学报》2016年第6期623-628,共6页Journal of Food Science and Biotechnology

基　　金：国家863计划项目(2012AA022103)

摘　　要：G.oxydans WSH-003能够高效地将D-山梨醇氧化为L-山梨糖,是工业VC生产的重要菌株。全基因组测序显示,该菌中存在3种不同的D-山梨醇脱氢酶基因簇(sldh),但这3种基因簇在菌株氧化D-山梨醇过程中的表达情况还未确定。WYLY HX,首先利用BLAST(Basic Local Alignment Search Tool)软件在线比对,确定了3种酶,分别为一种FAD(flavin adenine dinucleotide)辅酶依赖的山梨醇脱氢酶(FAD-SLDH)和两种不同的PQQ(pyrroloquinoline quinone)依赖的山梨醇脱氢酶(PQQ-SLDH)。然后,利用新一代高通量转录组测序技术,对发酵过程5个时期(4、8、12、20 h和40 h)的菌体分别进行转录组测序(RNA-Seq)。结果表明,基因座位为O1G_RS0104395/O1G_RS0104400的sld AB2基因在5个时期的转录水平FPKM(Fragments Per Kilobase of exon model per Million mapped reads)值均很低,而基因座位为O1G_RS0105425/O1G_RS0105430的sld AB1和基因座位为O1G_RS0106820/O1G_RS0106825/O1G_RS0106830的sld SLC基因的FPKM值较高,且都随着D-山梨醇的快速氧化而逐渐增大。研究结果表明,sld AB1和sld SLC基因为氧化葡萄糖酸杆菌WSH-003中主要表达的D-山梨醇脱氢酶基因。本研究成果为菌株中D-山梨醇脱氢酶的后续研发提供了方向。Gluconobacter oxydans WSH-003 is an L-sorbose production strain,mainly used of industrial production of Vitamin C. It can effectively oxidize D-sorbitol into L-sorbose with a high yield. The whole genome sequence of G. oxydans WSH-003 reveals that there are three different gene clusters for D-sorbitol dehydrogenase, whereas the gene expression of these three gene clusters in this strain has not been studied. In this study, two D-sorbitol dehydrogenases were proved to be PQQ dependent D-sorbitol dehydrogenase （PQQ-SLDH） and one was FAD dependent D-sorbitol dehydrogenase （FAD-SLDH） by using the BLAST software in NCBI. The transcription analysis of strain cells from 4 h, 8 h, 12 h, 20 h and 40 h in fermentation cultivation were further studied using the high through-out RNA-Seq technology. The result showed that sldhAB2 on gene locus O1G_RS0104395/O1G_RS0104400 had a very low transcript level （FPKM, Fragments Per Kilobase of exon model per Million mapped reads） in all 5 time points,compared with the high FPKM of sldhAB1 on gene locus O1G_RS0105425/O1G_RS0105430 and sldSLC on gene locus O1G_RS0106820/O1G_RS0106825 /O1G_RS0106830. Here, we illustrated that the gene cluster sldSLC and sldA BI played an important role in the oxidization of D-sorbitol to L-sorbose, which may contribute to the further study ofD-sorbitol dehydrogenase genes in the strain.

关 键 词：转录组测序 维生素C L-山梨糖 sldh基因簇 

分 类 号：Q78[生物学—分子生物学]