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PRODUCTION, PARTIAL CHARACTERIZATION, AND POTENTIAL DIAGNOSTIC USE OF SALICYLATE HYDROXYLASE FROM PSEUDOMONAS-PUTIDA UUC-1

Biomedical Sciences Research Institute Computer Science Research Institute Environmental Sciences Research Institute Nanotechnology & Advanced Materials Research Institute

Banat, Ibrahim, Marchant, A, Singh - Nee Nigam, Poonam, Gaston, SJS, Kelly, BA and Marchant, R (1994) PRODUCTION, PARTIAL CHARACTERIZATION, AND POTENTIAL DIAGNOSTIC USE OF SALICYLATE HYDROXYLASE FROM PSEUDOMONAS-PUTIDA UUC-1. ENZYME AND MICROBIAL TECHNOLOGY, 16 (8). pp. 665-670. [Journal article]

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Abstract

An unusual strain of Pseudomonas putida UUC-1 capable of growth at high salicylate concentration (10 gl(-1)) was investigated with the aim of developing an assay and a biosensor system for determining salicylate in body fluids by utilizing the salicylate hydroxylase enzyme. Medium and growth condition optimization were carried out under chemostat conditions. The highest biomass yield was at 4.0 gl(-1) salicylate, 25 degrees C, pH 6.5, and 0.2 h -1 dilution rate. Growth occurred at up to 0.45 h(-1) dilution rate, producing 236 Ul(-1) enzyme activity and an output of 424 Uh(-1) The activity and productivity were higher than any reported in the literature for this enzyme. It had a K-m value of 2.07 +/- 0.32 mu M and an M(r) of approximately 43,000. In addition its specific activity in the crude extract (0.8-0.9 U mg(-1) protein) was similar to the commercially available enzyme. No plasmid DNA was detected in this strain, and no salicylate-negative isolates were obtained when curing with mitomycin C. It is therefore proposed that our strain has a chromosomally located inducible salicylate hydroxylase gene that enables it to grow at high salicylate. This strain also offers a means of cheaply producing large quantities of the enzyme through standard fermentation techniques.

Item Type:Journal article
Faculties and Schools:Faculty of Life and Health Sciences
Faculty of Life and Health Sciences > School of Biomedical Sciences
Research Institutes and Groups:Biomedical Sciences Research Institute
Biomedical Sciences Research Institute > Infection and Immunity/Microbiology
Biomedical Sciences Research Institute > Pharmaceutical Science and Practice
ID Code:2801
Deposited By:Professor Roger Marchant
Deposited On:15 Dec 2009 10:25
Last Modified:16 May 2012 14:20

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