Abstract
Lipolytic enzymes from marine microbes have been the focus of intense and growing research. The bioluminescence bacterium Vibrio fischeri was produced esterase enzyme when the medium contained specific substrate. The esterase was purified from the concentrated culture supernatant. The most active fractions were obtained using the technique of precipitation with 1N HCl. The precipitated fraction was purified by ion exchange chromatography (DEAE-Cellulose) and gel filteration chromatography (Sephadex G200). The purified active fraction exhibiting final specific activity of 300U/mg and characterized; the optimum pH was 7.5, the optimum temperature was 30°C. The enzyme was very stable at the temperature 30°C and at wide range of pH. The enzyme was monomeric protein having molecular mass of 37 KDa estimated by native PAGE assay.
Highlights
Lipolytic enzymes from marine microbes have been the focus of intense and growing research
Material obtained in either of these ways was serially diluted in seawater complete (SWC) broth and the samples were spreaded on SWC (0.38M NaCl, 0.02M MgCl2.6H2O, 0.025M MgSO4.7H2O, 8mM KCl, 0.5% peptone, 0.3% yeast extract, 2% agar and 0.3% glycerol) agar plates and identified the luminous bacteria based on the work of Reichelt and Baumann (1973) and VHA (Vibrio Hareyi Agar) differential media (Harris et al, 1996)
Total viable luminous count range was varied from 4 to 18 CFU/ml. The habitats of this squid species must receive a significant input of cells of symbiotic V. fischeri (Lee and Ruby, 1994)
Summary
Lipolytic enzymes from marine microbes have been the focus of intense and growing research. The bioluminescence bacterium Vibrio fischeri was produced esterase enzyme when the medium contained specific substrate. The species is restricted to the marine environment and has a specific requirement for sodium ion for growth (Reichelt and Baumann, 1973). It occurs both free living in sea water (Ruby and Nealson, 1976) and as the specific luminous symbiont of the monocentrid fish and squid. The molecular and catalytic properties of this protein from mammalian sources have been well studied, only limited investigations have been made in to properties of isolated microbial esterases. The objective of this study is to focus on the esterase production of V. fischeri isolated from squid and on the purification and partial characterization of its esterase
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