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L-asparaginase from Bacillus flexus strain SS: Isolation, Screening, Production Process Optimization, Purification, and Anticancer Activity

DIR@IMTECH: CSIR-Institute of Microbial Technology

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Title L-asparaginase from Bacillus flexus strain SS: Isolation, Screening, Production Process Optimization, Purification, and Anticancer Activity
 
Creator Chand, S.
Mihooliya, K. N.
Sahoo, D. K.
Prasad, J. P.
Sharma, G.
 
Subject QR Microbiology
 
Description L-asparaginase has been widely accepted as a standard anticancer drug for acute lymphoblastic leukaemia (ALL). Presently in L-asparaginase biotherapeutic applications, the main focus is developing new L-asparaginase with minimal or without any glutaminase activity to reduce the associated adverse drug reactions. In this study, Bacillus flexus strain SS (NCBI GenBank Accession Number MN420983) was identified as a promising producer of L-asparaginase. L-asparaginase production was optimized by response surface methodology statistical bioprocess modelling, and enzyme yield of 25.08 IU/mL was reached at the bioreactor scale. The purification of L-asparaginase included ammonium sulfate precipitation, ion-exchange chromatography and size exclusion chromatography yielding 5.27-fold purification. The SDS-PAGE and CE-SDS revealed the monomeric L-asparaginase with molecular weight of 33 kDa. The purified enzyme was highly specific to substrate L-Asn and free from glutaminase activity. The anticancer activity of purified L-asparaginase was found specific against tumor cell lines SKBR3 (IC50 = 0.8 µg/mL), WIL2-S (IC50 = 16.2 µg/mL), and TF-1 (IC50 = 47 µg/mL), but not against negative control HUVEC cells. Therefore, L-asparaginase from B. flexus SS with no glutaminase activity could be a potential new candidate for anticancer drug for ALL with reduced adverse effects.
 
Publisher Springer
 
Date 2022-07-12
 
Type Article
PeerReviewed
 
Relation https://link.springer.com/article/10.1134/S0003683822040032
http://crdd.osdd.net/open/3055/
 
Identifier Chand, S. and Mihooliya, K. N. and Sahoo, D. K. and Prasad, J. P. and Sharma, G. (2022) L-asparaginase from Bacillus flexus strain SS: Isolation, Screening, Production Process Optimization, Purification, and Anticancer Activity. APPLIED BIOCHEMISTRY AND MICROBIOLOGY, 58 (4). pp. 416-427.