Background

Lipases play a crucial role in various industrial applications, and microbial lipases, particularly those from bacteria, possess significant properties. With increasing concerns about the environmental and health impacts of hydrocarbons from pipelines and refineries, there is a growing need to mitigate the risks associated with these compounds.

Methods

In this study, 40 bacterial isolates were recovered from contaminated soil samples collected from multiple refineries across Iraq. Using the Vitek system, bacterial isolates were identified up to the species level, revealing that only 12 isolates exhibited lipase-producing capabilities.

Results

Among the lipase-producing isolates, Ralstonia mannitolilytica demonstrated the highest extracellular lipase activity, as determined by an olive oil plate assay supplemented with rhodamine B. Confirmation of the species identity was achieved through 16S rRNA gene sequencing, with the obtained sequence deposited under accession number LC772176.1. Further sequence analysis revealed single nucleotide polymorphisms (SNPs) in the genome of Ralstonia mannitolilytica strain H230303-10_N19_7x_R2 (CP011257.1, positions 1,311,102 and 1,311,457). Additionally, the presence of the lipase gene was confirmed through amplification and sequencing using a thermocycler PCR. Sequence analysis of the gene, aligned using Geneious Prime software, identified SNPs (CP010799, CP049132, AY364601, CP011257, and CP023537), and a phylogenetic tree was constructed based on genetic characterization.

Conclusion

Our findings highlight the potential of Ralstonia mannitolilytica as a promising candidate for lipase production and contribute to our understanding of its genetic diversity and biotechnological applications in hydrocarbon degradation and industrial processes.

中文翻译：

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从伊拉克石油污染土壤中分离出的脂解甘露醇拉尔斯顿菌的表型和遗传特征

背景

脂肪酶在各种工业应用中发挥着至关重要的作用，微生物脂肪酶，特别是来自细菌的脂肪酶，具有重要的特性。随着人们越来越关注管道和炼油厂产生的碳氢化合物对环境和健康的影响，越来越需要减轻与这些化合物相关的风险。

方法

在这项研究中，从伊拉克各地炼油厂收集的受污染土壤样本中回收了 40 种细菌分离株。使用 Vitek 系统，对细菌分离株进行了物种水平的鉴定，结果表明只有 12 个分离株表现出脂肪酶生产能力。

结果

在产生脂肪酶的分离株中，解甘露醇罗尔斯顿菌 (Ralstonia mannitolilytica) 表现出最高的细胞外脂肪酶活性，通过添加罗丹明 B 的橄榄油平板测定法测定。通过 16S rRNA 基因测序确认物种身份，获得的序列以登录号保藏LC772176.1。进一步的序列分析揭示了解甘露醇罗尔斯顿菌菌株 H230303-10_N19_7x_R2 的基因组中的单核苷酸多态性 (SNP)（CP011257.1，位置 1,311,102 和 1,311,457）。此外，通过使用热循环仪 PCR 进行扩增和测序，证实了脂肪酶基因的存在。使用 Geneious Prime 软件对基因进行序列分析，鉴定出 SNP（CP010799、CP049132、AY364601、CP011257 和 CP023537），并根据遗传特征构建系统发育树。

结论

我们的研究结果强调了解甘露醇罗尔斯顿菌作为脂肪酶生产的有前途候选者的潜力，并有助于我们了解其遗传多样性以及在碳氢化合物降解和工业过程中的生物技术应用。