Study on production, purification and Characterisation of l-asparaginase from Escherichia coli and pseudomonas aeruginosa

Rai, Padmalatha S (2013) Study on production, purification and Characterisation of l-asparaginase from Escherichia coli and pseudomonas aeruginosa. International journal of pharmaceutical, chemical and biological sciences, 3 (3). pp. 565-570. ISSN 2249-9504

[img] PDF
sreepad.pdf - Published Version
Restricted to Registered users only

Download (253kB) | Request a copy

Abstract

Water sample microbes were isolated and screened for the potential producers of Lasparaginase using modified M-9 medium plate method. The single discrete colonies which have exhibited clear pink zone surrounding microbial colonies after 48 hours incubation indicate L-asparaginase producing cultures. This can easily be detected by the change in pH of the medium using phenol red. These colonies (L-Asparaginase positive cultures) were picked up and grown in the modified Czapeck Dox agar. E coli and Pseudomonas aeruginosa are optimised for comparative study and are inoculated in modified M-9 medium as production media. The largest quantity of L-asparaginase was produced when pseudomonas aeruginosa was grown in the modified M-9 medium for 52 hrs. Enzyme activity and specificity was calculated for all the crude and purified samples of the two different organisms. The maximum enzyme activity and specific activity was observed in Pseudomonas aeruginosa by using LAspargine as substrate. The activity of the Pseudomonas increased to 602 U/g as compare to specific activity of L-asparaginase from E coli 520 U/g. The molecular weight of the protein was found to be 75KDa. Characterisation of the enzyme showed optimum pH 7.0 for both E coli and Pseudomonas aeruginosa. Optimum temperature was 55`C.

Item Type: Article
Uncontrolled Keywords: Ammonium sulphate; Ion-exchange chromatography; modified M-9 medium.
Subjects: Life Sciences > MLSC Manipal
Depositing User: KMC Manipal
Date Deposited: 20 Jun 2014 10:00
Last Modified: 20 Jun 2014 10:00
URI: http://eprints.manipal.edu/id/eprint/139856

Actions (login required)

View Item View Item