Cloning, expression and characterization of a novel esterase (E29) from a marine bacterium Altererythrobacter luteolus SW109T

Abstract:

[Objective] We cloned, expressed and characterized a novel esterase E29 from a marine bacterium. [Methods] An assumed esterase gene, which was predicted via the genome of Altererythrobacter luteolus SW109^T and amplified by PCR, was cloned into expression vector pSMT3. The recombinant plasmid was transformed into Escherichia coli BL21(DE3). Subsequently, E29 was obtained via heterogenetic expression and characterized. [Results] The amino acids sequence analysis revealed E29 represents a new member of Family II of lipolytic enzyme. According to biochemical characterization, the maximum hydrolysis activity was obtained using p-nitrophenyl butyrate as substrate, at 45 ℃ and pH 8.5. The activity of E29 decreased in solution containing 10 mmol/L Co²⁺ or Mn²⁺, or 15% isopropanol or acetonitrile. The esterase was inactivated in 1% SDS solution. The addition of glycerol can promote the activity of E29 strikingly. [Conclusion] E29 is a novel marine esterase. Due to its high enzyme activity, wide substrates selectivity as well as tolerance of some organic solvents and metal irons, E29 has potential application in industry.