食品科学 ›› 2022, Vol. 43 ›› Issue (18): 166-175.doi: 10.7506/spkx1002-6630-20211126-326

• 生物工程 • 上一篇    

莱鲍迪甙C高效转化细菌Paenarthrobacter ilicis CR5301全基因组测序及关键糖苷酶分析

李洪飞,孙大庆,曹龙奎   

  1. (1.黑龙江八一农垦大学 国家杂粮工程技术中心,黑龙江 大庆 163319;2.东北石油大学化学化工学院,黑龙江 大庆 163318)
  • 发布日期:2022-09-28
  • 基金资助:
    黑龙江省普通本科高等学校青年创新人才培养计划项目(UNPYSCT-2017112); 牡丹江市科学技术计划项目(Z2017n0086;Z2018n010); 黑龙江八一农垦大学自然科学人才支持计划项目(ZRCPY202005); 黑龙江八一农垦大学学成、引进人才科研启动计划项目(XDB202009)

Whole Genome Sequencing and Key Glycosidase Analysis of Paenarthrobacter ilicis CR5301, an Efficient Rebaudioside C-converting Bacterium

LI Hongfei, SUN Daqing, CAO Longkui   

  1. (1. National Coarse Cereals Engineering Research Center, Heilongjiang Bayi Agricultural University, Daqing 163319, China; 2. College of Chemistry and Chemical Engineering, Northeast Petroleum University, Daqing 163318, China)
  • Published:2022-09-28

摘要: 为全面了解Paenarthrobacter ilicis CR5301的遗传背景,深入解析CR5301转化莱鲍迪甙C(rebaudioside C,RC)的关键酶,采用二代Illumina HiSeq和三代Nanopore相结合的测序方法,对CR5301进行全基因组测序和关键酶预测分析。结果显示,CR5301基因组为1 个闭合环状染色体DNA分子,不含质粒,基因组序列全长4 748 281 bp,GC含量62.92%,共编码4 458 个基因,同时含有4 个基因岛、1 个前噬菌体和14 个CRISPR-Cas编码序列。CR5301是P. ilicis物种第1个测定基因组完成图的菌株,也是Paenarthrobacter菌属已知基因组最大的菌株。基因组共线性分析和16S rRNA基因系统进化树分析结果一致表明,P. ilicis CR5301与P. aurescens具有更近的亲缘关系,而与P. ureafaciens的亲缘关系较远。7 个蛋白质数据库综合注释分析发现,P. ilicis CR5301基因组共含有523 个碳水化合物活性酶基因,其中18 个糖苷酶基因可能是CR5301转化RC的关键酶基因。最后,通过ProtParam、SOPMA生物信息学软件,预测了18 个糖苷酶的物化性质和二级结构。这些结果为CR5301的RC转化机制研究提供了清晰、完整的遗传信息,并且为P. ilicis物种广泛的生物学研究提供了完整、可靠的参考基因组序列,对今后P. ilicis物种生物学研究具有重要的参考价值和普遍的借鉴意义。

关键词: 微生物转化;莱鲍迪甙C;Paenarthrobacter ilicis CR5301;基因组;糖苷酶

Abstract: In order to comprehensively understand the genetic background of Paenarthrobacter ilicis CR5301 and dissect the key enzymes of CR5301 for transforming rebaudioside C (RC), the whole genome of CR5301 was sequenced using next-generation Illumina HiSeq and third-generation Nanopore-based DNA sequencing, and the key enzymes for RC convertion were predicted. The results showed that CR5301’s genome was a closed circular chromosomal DNA molecule without plasmids. The genome sequence was 4 748 281 bp in length, with a GC content of 62.92%. The genome encoded a total of 4 458 genes, including 4 gene islands, 1 prophage and 14 CRISPR-Cas coding sequences. CR5301 is the first strain of P. ilicis whose genome has been completely determined, and it is also the strain with the largest known genome of the genus Paenarthrobacter. P. ilicis CR5301 was more closely related to P. aurescens, but more distantly related to P. ureafaciens, according to genome collinearity and 16S rRNA gene-based phylogenetic tree analysis. Comprehensive annotation analysis against seven protein databases showed that P. ilicis CR5301’s genome contained 523 carbohydrate active enzyme genes, of which 18 glycosidase genes may be the key enzyme genes for the transformation of RC by CR5301. Finally, the physicochemical properties and secondary structures of the 18 glycosidases were predicted by the bioinformatics software ProtParam and SOPMA. These results provide clear and complete genetic information for understanding the mechanism of RC transformation by CR5301, and provide a complete and reliable reference genome sequence for extensive biological studies on P. ilicis.

Key words: microbial transformation; rebaudioside C; Paenarthrobacter ilicis CR5301; genome; glycosidases

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