Volume 20 No 20 (2022)
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Identification, PurificationAnd Insilico CharacterizationOf L-Glutaminase From Streptomyces Luteogriseus Isolated From Marine Isolate
Lavanya Kothapalli, Chandana Lakshmi M.V.V
Abstract
The hydrolytic conversion of l-glutamine to l-glutamate is carried out by glutaminases, a member of the large superfamily of serine-dependent beta-lactamases and penicillin-binding proteins. In this study, the purified Lglutaminases from Streptomyces luteogriseus isolated from marine sediments were purified and biochemically characterized. Neither d-glutamine nor l-asparagine were hydrolyzed by the purified glutaminase, which only exhibited strict L-glutamine specificity. L-glutaminase is an essential enzyme in the treatment of cancer when used in combination with other medications or drugs. Streptomyces luteogriseus, a newly isolated strain of Streptomyces species KLP-08, was found to be potentially producing extracellular L-glutaminase. The sequencing product was deposited in the GenBank database under the accession number ON623507. Using DEAE Sepharose CL-6B, ammonium sulfate precipitation, dialysis, and gel filtration chromatography, L-glutaminase was isolated from the crude enzyme for further characterization studies. For maximum L-glutaminase activity, the optimal pH, temperature, and incubation time were found to be 8.5, 40 °C, and 30 minutes, respectively. SDS–PAGE analysis revealed that the subunits of L-glutaminase have a molecular weight of approximately 50 kDa, making it the optimal substrate concentration. The L-glutaminase that was 3.38-fold purified had a final specific activity of 2.98U/mg protein. The production, purification, and characterization of L-glutaminase produced by Streptomyces luteogriseus, a newly isolated actinomycete from marine sediments, are summarized in greater detail for the first time in this study.
Keywords
Streptomyces luteogriseus, L-glutaminase, ammonium sulphate precipitation, dialysis, and gel filtration chromatography, Native PAGE, SDS-PAGE, 2D PAGE, MALDI-TOF/MS
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