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Purification and characterization of extracellular lipase from a thermotolerant strain: Bacillus subtilis TTP-06

DIR@IMTECH: CSIR-Institute of Microbial Technology

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Title Purification and characterization of extracellular lipase from a thermotolerant strain: Bacillus subtilis TTP-06
 
Creator Kaur , Manpreet
Kumar, Rakesh
Katoch, Poonam
Gupta, Reena
 
Subject QR Microbiology
 
Description In current study, lipase from a thermotolerant Bacillus subtilis TTP-06 was purified in a stepwise manner by using ammonium sulfate precipitation and column chromatography. Thenceforth, it was subjected to sodium dodecyl sulfate- and native-polyacrylamide gel electrophoresis to check the homogeneity of the purified enzyme. The ideal substrate concentration, pH, temperature, reaction duration and lipase specificity were identified. With a yield of 11.02%, purified lipase displayed activity of 8.51 U/mg. Thenceforward, the homogeneously purified enzyme was considered to be a homo-dimer of 30 kDa subunits. Enzyme had Km and Vmax value of 9.498 mM and 19.92 mol mg-1 min-1, respectively. Additionally, the matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) method was used to investigate the purified lipase and estimate its 3-D structure, which revealed a catalytic triad of serine, aspartate and histidine.
 
Publisher SPRINGER LINK
 
Date 2023-10
 
Type Article
PeerReviewed
 
Relation https://link.springer.com/article/10.1007/s13205-023-03717-6
http://crdd.osdd.net/open/3182/
 
Identifier Kaur , Manpreet and Kumar, Rakesh and Katoch, Poonam and Gupta, Reena (2023) Purification and characterization of extracellular lipase from a thermotolerant strain: Bacillus subtilis TTP-06. 3 Biotech, 13 (10). ISSN 2190-572X