Cloning, Characterization, and Structural Modeling of an Extremophilic Bacterial Lipase Isolated from Saline Habitats of
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Cloning, Characterization, and Structural Modeling of an Extremophilic Bacterial Lipase Isolated from Saline Habitats of the Thar Desert Swati Verma 1 & Rajender Kumar 2 & Pradeep Kumar 3 & Deepak Sharma 3 & Hukam Gahlot 4 & Pushpender Kumar Sharma 5 & Gautam Kumar Meghwanshi 1 Received: 30 December 2019 / Accepted: 23 April 2020/ # Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract
Lipases have a characteristic folding pattern of α/β-hydrolase with mostly parallel βsheets, flanked on both sides by α-helixes in the structure. The active site is formed by a catalytic triad (serine, aspartic/glutamic acid, and histidine), which is highly conserved. In this study, we have used an integrated experimental and computational approach to identify the extremophilic microbial lipases from the saline habitats of the Thar Desert of Rajasthan. Lipase-producing bacteria were screened and a few samples showed significant lipase activity in both quantitative and qualitative experiments. 16S rRNA sequence analysis of the isolate F1 showed that its sequence is quite similar to that of Bacillus licheniformis and Bacillus haynesii, indicating that this isolate belongs to a new subspecies of Bacillus. The isolate F7 showed maximum sequence identity with Bacillus tequilensis strain 10b. The isolate F7 sequence analysis provided a clear testimony that it can be a new strain of Bacillus tequilensis. The F7 lipase exhibited optimal activity at 60 °C and pH 9. Structural modeling of the F7 lipase revealed that it has a highly conserved alpha/beta hydrolase fold at the sequence and structural level except for the Nterminal region. Interestingly, residue Glu128 was different from the template structure and showed the hydrogen bonding between the side chain of Glu128 and side chains of Asn35 and Gln152 amino acids. Besides, this amino acid also showed salt bridge interaction between Glu128–Lys101. These interactions may be assisting in preserving the stability and activity of lipase at high temperatures and in alkaline pH conditions. The information gathered from this investigation will guide in the rational designing of new more potential extremophilic lipase. Keywords Lipases . Extremophilic . Thar Desert . Arid regions . Bacillus tequilensis . Molecular modeling . Signal peptide . Thermoalkaliphilic Electronic supplementary material The online version of this article (https://doi.org/10.1007/s12010-02003329-3) contains supplementary material, which is available to authorized users.
* Gautam Kumar Meghwanshi [email protected] Extended author information available on the last page of the article
Applied Biochemistry and Biotechnology
Introduction Lipases (triacylglycerol acyl hydrolase, EC 3.1.1.3) are ubiquitous enzymes that catalyze the cleavage of ester bonds of fats (oils, animal fats, etc.) as well as other synthetic esters in the presence of water, and subsequent release of diacylglycerols, monoacylglycerols, free fatty acids, and glycerol in case of fats and other products in case of synthetic ester
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