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Development of molecular driven screening for desulfurizing microorganisms targeting the dszB desulfinase gene
Research in Microbiology ( IF 2.5 ) Pub Date : 2021-08-08 , DOI: 10.1016/j.resmic.2021.103872
Emmanuel Duval 1 , Cristiana Cravo-Laureau 2 , Line Poinel 3 , Robert Duran 2
Affiliation  

COnsensus DEgenerate Hybrid Oligonucleotide Primers (CODEHOP) were developed for the detection of the dszB desulfinase gene (2′-hydroxybiphenyl-2-sulfinate desulfinase; EC 3.13.1.3) by polymerase chain reaction (PCR), which allow to reveal larger diversity than traditional primers. The new developed primers were used as molecular monitoring tool to drive a procedure for the isolation of desulfurizing microorganisms. The primers revealed a large dszB gene diversity in environmental samples, particularly in diesel-contaminated soil that served as inoculum for enrichment cultures. The isolation procedure using the dibenzothiophene sulfone (DBTO2) as sole sulfur source reduced drastically the dszB gene diversity. A dszB gene closely related to that carried by Gordonia species was selected. The desulfurization activity was confirmed by the production of desulfurized 2-hydroxybiphenyl (2-HBP). Metagenomic 16S rRNA gene sequencing showed that the Gordonia genus was represented at low abundance in the initial bacterial community. Such observation highlighted that the culture medium and conditions represent the bottleneck for isolating novel desulfurizing microorganisms. The new developed primers constitute useful tool for the development of appropriate cultural-dependent procedures, including medium and culture conditions, to access novel desulfurizing microorganisms useful for the petroleum industry.



中文翻译:

开发针对 dszB 脱硫酶基因的脱硫微生物的分子驱动筛选

开发了一致的简并杂合寡核苷酸引物 (CODEHOP),用于通过聚合酶链式反应 (PCR) 检测dsz B 脱硫酶基因 (2'-羟基联苯-2-亚磺酸脱硫酶;EC 3.13.1.3),它可以揭示比聚合酶链反应更大的多样性。传统引物。新开发的引物被用作分子监测工具来驱动分离脱硫微生物的程序。引物揭示了环境样品中的大量dsz B 基因多样性,特别是在用作富集培养物的柴油污染土壤中。使用二苯并噻吩砜 (DBTO 2 ) 作为唯一硫源的分离程序大大降低了dsz B 基因的多样性。一个dsz选择与Gordonia物种携带的B基因密切相关的B基因。脱硫活性通过脱硫的 2-羟基联苯 (2-HBP) 的生成得到证实。宏基因组 16S rRNA 基因测序显示Gordonia属在初始细菌群落中的丰度较低。这样的观察突出表明,培养基和条件是分离新型脱硫微生物的瓶颈。新开发的引物构成了开发适当的培养依赖程序(包括培养基和培养条件)的有用工具,以获取对石油工业有用的新型脱硫微生物。

更新日期:2021-10-17
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