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从秘鲁尾矿中分离出的 的基因组特征及其氰化物降解酶 CynD 的评估。

Genomic Characterization of Isolated from Mine Tailings in Peru and Evaluation of Its Cyanide-Degrading Enzyme CynD.

机构信息

Facultad de Ciencias Biológicas, Universidad Ricardo Palmagrid.441904.c, Lima, Peru.

Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, Brazil.

出版信息

Appl Environ Microbiol. 2022 Jul 26;88(14):e0091622. doi: 10.1128/aem.00916-22. Epub 2022 Jun 28.

Abstract

Understanding the biochemistry and metabolic pathways of cyanide degradation is necessary to improve the efficacy of cyanide bioremediation processes and industrial requirements. We have isolated and sequenced the genome of a cyanide-degrading strain from water in contact with mine tailings from Lima, Peru. This strain was classified as Bacillus safensis based on 16S rRNA gene sequencing and core genome analyses and named B. safensis PER-URP-08. We searched for possible cyanide-degradation enzymes in the genome of this strain and identified a putative cyanide dihydratase (CynD) gene similar to a previously characterized CynD from Bacillus pumilus C1. Sequence analysis of CynD from and allow us to identify C-terminal residues that differentiate both CynDs. We then cloned, expressed in Escherichia coli, and purified recombinant CynD from PER-URP-08 (CynD) and showed that in contrast to CynD from C1, this recombinant CynD remains active at up to pH 9. We also showed that oligomerization of CynD decreases as a function of increased pH. Finally, we demonstrated that transcripts of CynD in PER-URP-08 are strongly induced in the presence of cyanide. Our results suggest that the use of PER-URP-08 and CynD as potential tool for cyanide bioremediation warrants further investigation. Despite being of environmental concern around the world due to its toxicity, cyanide continues to be used in many important industrial processes. Thus, searching for cyanide bioremediation methods is a matter of societal concern and must be present on the political agenda of all governments. Here, we report the isolation, genome sequencing and characterization of cyanide degradation capacity of a bacterial strain isolated from an industrial mining site in Peru. We characterize a cyanide dehydratase (CynD) homolog from one of these bacteria, Bacillus safensis PER-URP-08.

摘要

了解氰化物降解的生物化学和代谢途径对于提高氰化物生物修复过程的效率和工业需求是必要的。我们已经从秘鲁利马的尾矿接触水中分离并测序了一种能够降解氰化物的菌株的基因组。该菌株基于 16S rRNA 基因测序和核心基因组分析被分类为 Bacillus safensis,并命名为 B. safensis PER-URP-08。我们在该菌株的基因组中搜索可能的氰化物降解酶,并鉴定出一个类似先前从 Bacillus pumilus C1 中鉴定出的氰化物二水合酶(CynD)的推定基因。对 和 中 CynD 的序列分析使我们能够识别区分这两种 CynD 的 C 末端残基。然后,我们克隆、在大肠杆菌中表达并纯化了来自 PER-URP-08 的重组 CynD(CynD),并表明与来自 C1 的 CynD 不同,该重组 CynD 在高达 pH 9 的条件下仍保持活性。我们还表明,随着 pH 值的增加,CynD 的寡聚化程度降低。最后,我们证明了 PER-URP-08 中的 CynD 转录物在存在氰化物的情况下强烈诱导。我们的结果表明,PER-URP-08 和 CynD 的使用作为氰化物生物修复的潜在工具值得进一步研究。尽管氰化物由于其毒性而在全世界引起关注,但它仍在许多重要的工业过程中使用。因此,寻找氰化物生物修复方法是一个关乎社会的问题,必须成为所有政府政治议程的一部分。在这里,我们报告了从秘鲁一个工业采矿场分离的一株细菌的氰化物降解能力的分离、基因组测序和表征。我们从这些细菌之一的 Bacillus safensis PER-URP-08 中鉴定出一种氰化物脱水酶(CynD)同源物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0404/9317851/7660807ddfac/aem.00916-22-f001.jpg

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