Pacawadee TirawongsarojRutchadaporn SriprangPiyanun HarnpicharnchaiTaksawan ThongaramVerawat ChampredaSutipa TanapongpipatKusol PootanakitLily EurwilaichitrThailand National Center for Genetic Engineering and BiotechnologyMahidol University2018-07-122018-07-122008-01-01Journal of Biotechnology. Vol.133, No.1 (2008), 42-49016816562-s2.0-36249002945https://repository.li.mahidol.ac.th/handle/20.500.14594/18994Functional screening for lipolytic enzymes from a metagenomic library (origin: Jae Sawn hot spring, Thailand) resulted in isolation of a novel patatin-like phospholipase (PLP) and an esterase (Est1). PLP contained four conserved domains similar to other patatin-like proteins with lipid acyl hydrolase activity. Likewise, sequence alignment analysis revealed that Est1 can be classified as a family V bacterial lipolytic enzyme. Both PLP and Est1 were expressed heterologously as soluble proteins in E. coli and exhibited more than 50% of their maximal activities at alkaline pH, of 7-9 and 8-10, respectively. In addition, both enzymes retained more than 50% of maximal activity in the temperature range of 50-75 °C, with optimal activity at 70 °C and were stable at 70 °C for at least 120 min. Both PLP and Est1 exhibited high Vmaxtoward p-nitrophenyl butyrate. The enzymes had activity toward both short-chain (C4and C5) and long chain (C14and C16) fatty acid esters. The isolated enzymes, are therefore, different from other known patatin-like phospholipases and esterases, which usually show no activity for substrates longer than C10. We suggest that PLP and EstA enzymes are novel and have a; b potential use in industrial applications. © 2007 Elsevier B.V. All rights reserved.Mahidol UniversityBiochemistry, Genetics and Molecular BiologyArticleSCOPUS10.1016/j.jbiotec.2007.08.046