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
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Analysis of D-Sorbitol Dehydrogenase Gene in Gluconobacter oxydans WSH-003
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    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sldAB2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sldAB1和基因座位为O1G_RS0106820/O1G_RS0106825 /O1G_RS0106830的sldSLC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,sldAB1和sldSLC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。

    Abstract:

    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 sldAB1 played an important role in the oxidization of D-sorbitol to L-sorbose,which may contribute to the further study of D-sorbitol dehydrogenase genes in the strain.

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胡于东,堵国成,陈坚,周景文.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[J].食品与生物技术学报,2016,35(6):623-628.

HU Yudong, DU Guocheng, CHEN Jian, ZHOU Jingwen. Analysis of D-Sorbitol Dehydrogenase Gene in Gluconobacter oxydans WSH-003[J]. Journal of Food Science and Biotechnology,2016,35(6):623-628.

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  • 在线发布日期: 2016-11-02
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