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Studies on efficient production of a novel l-asparaginase by a newly isolated Pseudomonas resinovoransIGS-131 and its heterologous expression in Escherichia coli

Authors :
Mihooliya, Kanti N.
Nandal, Jitender
Kumari, Alka
Nanda, Sidhanta
Verma, Himanshu
Sahoo, Debendra K.
Source :
3 Biotech; April 2020, Vol. 10 Issue: 4
Publication Year :
2020

Abstract

In the current study, the production of novel glutaminase free l-asparaginase from a new microbial source (Pseudomonas resinovoransIGS-131) is reported. Optimization of l-asparaginase production using conventional and statistical optimization techniques resulted in an enzyme yield of 37.63 IU/mL, which was 3.45-fold higher than the initial enzyme activity (i.e., 10.91 IU/mL). l-Asparaginase production from P. resinovoransIGS-131 was successfully carried out at the bioreactor level and investigations on the effect of agitation rates showed a maximum asparaginase yield of 38.88 IU/mL after 24 h fermentation at 400 rpm. The l-asparaginase gene from this source, showing 78% identity with a reported sequence in GenBank, was expressed in Escherichia colirosetta DE3. The molecular weight of the recombinant protein was determined as 35.6 kDa. Downstream processing of recombinant l-asparaginase resulted in a purified protein concentration of 62.53 mg/L, which showed good free radical scavenging activity of 62%. The current findings provide promising results for a process of l-asparaginase production from P. resinovoransIGS-131. Furthermore, the recombinant production of this enzyme could help in avoiding the complexity of down streaming processes associated with the purification of this enzyme from wild-type organisms.

Details

Language :
English
ISSN :
2190572X and 21905738
Volume :
10
Issue :
4
Database :
Supplemental Index
Journal :
3 Biotech
Publication Type :
Periodical
Accession number :
ejs52570755
Full Text :
https://doi.org/10.1007/s13205-020-2135-4