title: Substitution of Val72 residue alters the enantioselectivity and activity of Penicillium expansum lipase.
abstract: Error-prone PCR was used to create more active or enantioselective variants of Penicillium expansum lipase (PEL). A variant with a valine to glycine substitution at residue 72 in the lid structure exhibited higher activity and enantioselectivity than those of wild-type PEL. Site-directed saturation mutagenesis was used to explore the sequence-function relationship and the substitution of Val72 of P. expansum lipase changed both catalytic activity and enantioselectivity greatly. The variant V72A, displayed a highest enantioselectivity enhanced to about twofold for the resolution of (R, S)-naproxen (E value increased from 104 to 200.7 for wild-type PEL and V72A variant, respectively). In comparison to PEL, the variant V72A showed a remarkable increase in specific activity towards p-nitrophenyl palmitate (11- and 4-fold increase at 25 and 35 °C, respectively) whereas it had a decreased thermostability. The results suggest that the enantioselective variant V72A could be used for the production of pharmaceutical drugs such as enantiomerically pure (S)-naproxen and the residue Val 72 of P. expansum lipase plays a significant role in the enantioselectivity and activity of this enantioselective lipase.
authors.affiliation: College of Life Sciences, Fujian Normal University, Fuzhou 350108, China. biotlh@gmail.com
authors.forename: Lianghua
authors.initials: L
authors.lastname: Tang
authors.forename: Min
authors.initials: M
authors.lastname: Su
authors.forename: Ling
authors.initials: L
authors.lastname: Zhu
authors.forename: Liying
authors.initials: L
authors.lastname: Chi
authors.forename: Junling
authors.initials: J
authors.lastname: Zhang
authors.forename: Qiong
authors.initials: Q
authors.lastname: Zhou
mesh_terms: D019943:Amino Acid Substitution; D008049:Lipase / Q000235:genetics* / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed / Q000379:methods; D009288:Naproxen / Q000494:pharmacology; D010168:Palmitates / Q000378:metabolism; D010407:Penicillium / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D013329:Structure-Activity Relationship; D013379:Substrate Specificity
chemical_list: D010168:Palmitates; D009288:Naproxen; D008049:Lipase