Isolation and Characterization of a Family VII Esterase Derived from Alluvial Soil Metagenomic Library

Weixin Tao; Myung Hwan Lee; Jing Wu; Nam Hee Kim; Seon-Woo Lee

doi:10.1007/s12275-011-1102-5

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Isolation and Characterization of a Family VII Esterase Derived from Alluvial Soil Metagenomic Library: Weixin Tao ¹, Myung Hwan Lee ², Jing Wu ², Nam Hee Kim ², Seon-Woo Lee ^1,2; Journal of Microbiology 2011;49(2):178-185.
DOI: https://doi.org/10.1007/s12275-011-1102-5
Published online: May 3, 2011

¹Department of Medical Bioscience, Dong-A University, Busan 604-714, Republic of Korea, ²Department of Applied Biology, Dong-A University, Busan 604-714, Republic of Korea

Corresponding author: Seon-Woo Lee , Tel: +82-51-200-7551,

Received: 3 March 2011 • Accepted: 6 April 2011

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Abstract

A novel esterase gene, estDL30, was isolated from an alluvial metagenomic library using function-driven screening. estDL30 consisted of 1,524 nucleotides and encoded a 507-amino acid protein. Sequence analysis revealed that EstDL30 is similar to many type B carboxylesterases, containing a G-E-S-A-G pentapeptide with a catalytic Ser residue. Phylogenetic analysis suggested that EstDL30 belongs to the family VII lipases, together with esterases from Bacillus subtilis (P37967), Streptomyces coelicolor A3(2) (CAA22794), and Arthrobacter oxydans (Q01470). Purified EstDL30 showed its highest catalytic efficiency toward p-nitrophenyl butyrate, with a kcat of 229.3 s-1 and kcat/Km of 176.4 s-1mM-1; however, little activity was detected when the acyl chain length exceeded C8. Biochemical characterization of EstDL30 revealed that it is an alkaline esterase that possesses maximal activity at pH 8 and 40°C. The effects of denaturants and divalent cations were also investigated. EstDL30 tolerated well the presence of methanol and Tween 20. Its activity was strongly inhibited by 1 mM Cu2+ and Zn2+, but stimulated by Fe2+. The unique properties of EstDL30, its high activity under alkaline conditions and stability in the presence of organic solvents, may render it applicable to organic synthesis.

Keywords: alluvial soil;esterase;family VII lipase;metagenome

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Isolation and Characterization of a Family VII Esterase Derived from Alluvial Soil Metagenomic Library

J. Microbiol. 2011;49(2):178-185. Published online May 3, 2011

DOI: https://doi.org/10.1007/s12275-011-1102-5

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