Loop grafting of Bacillus subtilis lipase A: inversion of enantioselectivity

Chemistry & Biology
Ykelien L BoersmaWim J Quax

Abstract

Lipases are successfully applied in enantioselective biocatalysis. Most lipases contain a lid domain controlling access to the active site, but Bacillus subtilis Lipase A (LipA) is a notable exception: its active site is solvent exposed. To improve the enantioselectivity of LipA in the kinetic resolution of 1,2-O-isopropylidene-sn-glycerol (IPG) esters, we replaced a loop near the active-site entrance by longer loops originating from Fusarium solani cutinase and Penicillium purpurogenum acetylxylan esterase, thereby aiming to increase the interaction surface for the substrate. The resulting loop hybrids showed enantioselectivities inverted toward the desired enantiomer of IPG. The acetylxylan esterase-derived variant showed an inversion in enantiomeric excess (ee) from -12.9% to +6.0%, whereas the cutinase-derived variant was improved to an ee of +26.5%. The enantioselectivity of the cutinase-derived variant was further improved by directed evolution to an ee of +57.4%.

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Citations

Sep 6, 2013·Journal of Industrial Microbiology & Biotechnology·Hong Seung YunYoung Je Yoo
Nov 4, 2011·International Journal of Phytoremediation·Joseph M Jez
Jun 14, 2012·Protein Engineering, Design & Selection : PEDS·Humberto FloresEnrique Morett
Jan 15, 2013·Journal of Biotechnology·Nagihan AkbulutFüsun Gümüşel
Nov 29, 2013·Biotechnology and Bioengineering·Hyun Tae HwangArvind Varma
Apr 24, 2012·Chemistry & Biology·Moshe GoldsmithDan S Tawfik
Jan 1, 2012·Computational and Structural Biotechnology Journal·Arnau BassegodaPilar Diaz
Jul 2, 2010·Chembiochem : a European Journal of Chemical Biology·Robert KouristUwe T Bornscheuer
Nov 24, 2011·Chembiochem : a European Journal of Chemical Biology·Jan G E van LeeuwenDick B Janssen
Nov 25, 2021·Environmental Microbiology Reports·Mousumi BhattacharyyaTapan K Dutta

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