背景:脂肪酶在各种工业应用中起着至关重要的作用,和微生物脂肪酶,尤其是细菌,具有显著的属性。随着人们越来越关注管道和炼油厂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:在本研究中,从伊拉克多个炼油厂收集的污染土壤样品中回收了40种细菌分离物。使用维泰克系统,细菌分离株被鉴定到物种水平,揭示只有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物中,甘露醇Ralstonia表现出最高的细胞外脂肪酶活性,如通过补充了罗丹明B的橄榄油板测定所确定的。通过16SrRNA基因测序实现了物种身份的确认,获得的序列保存在登录号LC772176.1下。进一步的序列分析显示,甘露醇Ralstonia菌株H230303-10_N19_7x_R2(CP011257.1,位置1,311,102和1,311,457)基因组中的单核苷酸多态性(SNP)。此外,通过使用热循环仪PCR进行扩增和测序来确认脂肪酶基因的存在.基因的序列分析,使用GeneiousPrime软件对齐,经鉴定的SNP(CP010799、CP049132、AY364601、CP011257和CP023537),并根据遗传特征构建了系统进化树。
结论:我们的发现强调了甘露糖醇作为脂肪酶生产的有希望的候选者的潜力,并有助于我们了解其遗传多样性和在碳氢化合物降解和工业过程中的生物技术应用。
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.
CONCLUSIONS: 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.