Saleem Farooq, Shabir Ahmad Ganai, Bashir Ahmad Ganai, Suma Mohan, Baba Uqab, Ruqeya Nazir
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引用次数: 6
Abstract
Lipases from Pseudomonas species are particularly useful due to their broader biocatalytic applications and temperature activity. In this study, we amplified the gene encoding wild-type cold-active lipase from the genome of psychrotrophic bacterium isolated from the Himalayan glacier. The isolated CRBC14 strain was identified as Pseudomonas sp. based on the 16S rRNA gene sequence. Lipase activity was determined by observing the hydrolysis zone on nutrient agar containing tributyrin (1%, v/v). The sequence analysis of cold-active lipase revealed a protein of 611 amino acids with a calculated molecular mass of 63.71 kDa. The three-dimensional structure of this lipase was generated through template-supported modeling. Distinct techniques stamped the model quality, following which the binding free energies of tributyrin and oleic acid in the complex state with this enzymatic protein were predicted through molecular mechanics generalized born surface area (MMGBSA). A relative comparison of binding free energy values of these substrates indicated tributyrin's comparatively higher binding propensity towards the lipase. Using molecular docking, we evaluated the binding activity of cold-active lipase against tributyrin and oleic acid. Our docking analysis revealed that the lipase had a higher affinity for tributyrin than oleic acid, as evidenced by our measurement of the hydrolysis zone on two media plates. This study will help to understand the bacterial diversity of unexplored Himalayan glaciers and the possible application of their cold-adapted enzymes.
期刊介绍:
Current Genetics publishes genetic, genomic, molecular and systems-level analysis of eukaryotic and prokaryotic microorganisms and cell organelles. All articles are peer-reviewed. The journal welcomes submissions employing any type of research approach, be it analytical (aiming at a better understanding), applied (aiming at practical applications), synthetic or theoretical.
Current Genetics no longer accepts manuscripts describing the genome sequence of mitochondria/chloroplast of a small number of species. Manuscripts covering sequence comparisons and analyses that include a large number of species will still be considered.