High level expression and characterization of a novel thermostable, organic solvent tolerant, 1,3-regioselective lipase from Geobacillus sp. strain ARM

Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia.
Bioresource Technology (Impact Factor: 5.04). 07/2011; 102(13):6972-81. DOI: 10.1016/j.biortech.2011.03.083
Source: PubMed

ABSTRACT The mature ARM lipase gene was cloned into the pTrcHis expression vector and over-expressed in Escherichia coli TOP10 host. The optimum lipase expression was obtained after 18 h post induction incubation with 1.0mM IPTG, where the lipase activity was approximately 1623-fold higher than wild type. A rapid, high efficient, one-step purification of the His-tagged recombinant lipase was achieved using immobilized metal affinity chromatography with 63.2% recovery and purification factor of 14.6. The purified lipase was characterized as a high active (7092 U mg(-1)), serine-hydrolase, thermostable, organic solvent tolerant, 1,3-specific lipase with a molecular weight of about 44 kDa. The enzyme was a monomer with disulfide bond(s) in its structure, but was not a metalloenzyme. ARM lipase was active in a broad range of temperature and pH with optimum lipolytic activity at pH 8.0 and 65°C. The enzyme retained 50% residual activity at pH 6.0-7.0, 50°C for more than 150 min.

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Available from: Afshin Ebrahimpour, May 16, 2014
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    • "Many genes of thermophilic enzymes have been isolated in recent years from the genomes of Geobacillus for their further heterological expression in mesophylic bacteria. Among these enzymes, lipases are found (AbdellFatt tah, Gaballa, 2008; QuintanaaCastro et al., 2009; Cheong et al., 2011; Ebrahimpour et al., 2011; Balan et al., 2012). They take part in the decomposition of lignocellulosic biomass: βglucosidase (Shallom et al., RUSSIAN JOURNAL OF GENETICS: APPLIED RESEARCH Vol. 4 No. 3 2014 2005; BenDavid et al., 2007; Wagschal et al., 2009; Ratnadewi et al., 2013), endoglucanase (Ng et al., 2009), xylanase (Wu et al., 2006; Canakci et al., 2007, 2012; Gerasimova, Kuisiene, 2012; Liu et al., 2012; Verma et al., 2013), and many other proteins. "
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