食品科学 ›› 2024, Vol. 45 ›› Issue (22): 73-84.doi: 10.7506/spkx1002-6630-20240228-147

• 生物工程 • 上一篇    下一篇

产褐藻胶裂解酶菌株S10的鉴定、全基因组测序及分析

刘芳, 舒志强, 王共明, 井月欣, 赵云苹, 徐英江, 矫春娜, 张健   

  1. (1.山东省海洋资源与环境研究院,烟台市海珍品质量安全控制与精深加工重点实验室,山东 烟台 264006;2.上海海洋大学食品学院,上海 201306)
  • 出版日期:2024-11-25 发布日期:2024-11-05
  • 基金资助:
    山东省现代农业产业技术体系藻类产业创新团队建设项目(SDAIT-26-05); 山东省自然科学基金重点项目(ZR2020KC034)

Identification, Whole-Genome Sequencing and Analysis of Vibrio alginolyticus S10, a Strain Producing Alginate Lyase

LIU Fang, SHU Zhiqiang, WANG Gongming, JING Yuexin, ZHAO Yunping, XU Yingjiang, JIAO Chunna, ZHANG Jian   

  1. (1. Yantai Key Laboratory of Quality and Safety Control and Deep Processing of Marine Food, Shandong Marine Resource and Environment Research Institute, Yantai 264006, China; 2. College of Food, Shanghai Ocean University, Shanghai 201306, China)
  • Online:2024-11-25 Published:2024-11-05

摘要: 为了详细解析从仿刺参肠道中分离出1 株具有较高酶活力高产褐藻胶裂解酶菌株S10的基因组序列信息,进而深入挖掘与褐藻胶裂解酶相关的基因资源,本研究通过形态学观察和16S rRNA序列分析对菌株S10进行菌种鉴定,然后利用Illumina二代测序技术和第三代高通量PacBio测序平台对菌株S10进行全基因组测序,并使用相关软件对测序数据进行基因组组装、基因预测及基因功能注释等。此外,根据注释结果对菌株S10中所含的3 组假定褐藻胶裂解酶基因序列进行生物信息学分析及结构预测。结果表明,菌株S10被鉴定为Vibrio alginolyticus,基因组总长度为5 397 046 bp,GC含量为44.59%,由2 条染色体和1 条质粒组成。预测共有4 936 个编码基因,127 个tRNA基因和37 个rRNA基因。在直系同源集、基因本体论、京都基因与基因组百科全书和碳水化合物活性酶数据库中分别注释到4 039、3 163、3 104 个和96 个功能基因。此外,在菌株S10中发现了3 组潜在的褐藻胶裂解酶基因alg4755、alg4756和alg4760。生物信息学分析结果表明,褐藻胶裂解酶Alg4755、Alg4756和Alg4760均属于多糖裂解酶家族7(polysaccharide lyases,PL7)蛋白,具有3 个PL7家族高度保守的基序(R*E*R、Q(I/V)H、Y*KAG*Y*Q)。综上,S10菌株全基因组测序及分析对该菌的高效产酶机制研究和新型褐藻胶裂解酶的挖掘具有重要意义,可为后期酶的表达及工业化应用提供理论基础。

关键词: 褐藻胶裂解酶;溶藻弧菌;全基因组测序;基因功能注释;生物信息学

Abstract: Strain S10, isolated from the intestine of Apostichopus japonicus, is able to produce alginate lyase at a high yield. In order to scrutinize its genome sequence and to explore gene resources related to alginate lyase, strain S10 was identified through morphological observation and 16S rRNA sequence analysis. Then, its whole genome was sequenced using Illumina second-generation sequencing technology and PacBio, the third-generation high-throughput sequencing platform. The sequencing data were used for genome assembly, gene prediction, and gene function annotation. In addition, based on the annotation results, bioinformatics analysis and structure prediction were performed on three sets of putative alginate lyase gene sequences in strain S10. The results showed that strain S10 was identified as Vibrio alginolyticus, with a total genome length of 5 397 046 bp and a GC content of 44.59%. The genome was composed of two chromosomes and one plasmid. A total of 4 936 protein-coding genes, 127 tRNA genes and 37 rRNA genes were predicted. Totally 4 039, 3 163, 3 104 and 96 functional genes were annotated in the Clusters of Orthologous Groups (COG), Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) and Carbohydrate-Active EnZymes (CAZy) databases, respectively. In addition, three groups of potential alginate lyase genes, alg4755, alg4756 and alg4760, were found in strain S10. Bioinformatics analysis showed that alginate lyases Alg4755, Alg4756 and Alg4760 all belonged to the polysaccharide lyase family 7 (PL7) and had three highly conserved motifs of the PL7 family, R*E*R, Q(I/V)H and Y*KAG*Y*Q. In summary, the sequencing and analysis of the whole genome of S10 strain is of great significance for understanding the mechanism of efficient enzyme production by this strain and for exploring new alginate lyases, and also provides a theoretical basis for follow-up research on enzyme expression and industrial application.

Key words: alginate lyase; Vibrio alginolyticus; whole genome sequencing; gene functional annotation; bioinformatics

中图分类号: