Gel-immobilized enzymes as promising biocatalysts: Results from Indo-Russian collaborative studies

Markvicheva, Elena A.; Lozinsky, Vladimir I.; Plieva, Fatima M.; Kochetkov, Konstantin A.; Rumsh, Lev D.; Zubov, Vitali P.; Maity, Jyotirmoy; Kumar, Rajesh; Parmar, Virinder S.; Belokon, Yury N.

doi:10.1351/pac200577010227

Pure Appl. Chem., 2005, Vol. 77, No. 1, pp. 227-236

http://dx.doi.org/10.1351/pac200577010227

Gel-immobilized enzymes as promising biocatalysts: Results from Indo-Russian collaborative studies

Elena A. Markvicheva¹, Vladimir I. Lozinsky², Fatima M. Plieva², Konstantin A. Kochetkov², Lev D. Rumsh¹, Vitali P. Zubov¹, Jyotirmoy Maity³, Rajesh Kumar³, Virinder S. Parmar³ and Yury N. Belokon²

¹ Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya St. 16/10, Moscow 117871, Russia
² Nesmeyanov Institute of Organoelement Compounds, Russian Academy of Sciences, Vavilov St. 28, Moscow, V-334, 117813, Russia
³ Bioorganic Laboratory, Department of Chemistry, University of Delhi, Delhi 110 007, India

Abstract: Chemo-enzymatic methods constitute a promising approach to obtain various biologically active compounds, including enantiomerically pure substances. Entrapment in gels is one of the most convenient methods to stabilize enzymes for their application in water/organic media. Proteases and lipases are widely used for enantioselective transformations of various organic compounds in water-poor media. In this study, chymotrypsin was entrapped into a composite poly(N-vinyl caprolactam)-calcium alginate (PVCL-CaAlg) and covalently attached to poly(vinyl alcohol) (PVA) cryogel beads. Lipase was immobilized by covalently attaching to aldehyde-bearing PVA cryogel beads. The activities of the entrapped biocatalysts were studied. Both entrapped α-chymotrypsin and lipase retained high activity in acetonitrile/water medium (water content 0.5–20 %) and displayed high storage stability for several months. The high operational stability of immobilized α-chymotrypsin and lipase in a cyclic process (up to 912 h in total) was also demonstrated. Gel-immobilized enzymes were successfully used to obtain optically pure L-phenylalanine (ee 98.6 and 83 % in the case of α‑chymotrypsin and lipase, respectively) by enantioselective hydrolysis of Schiff’s base of amino acid ethyl ester in an acetonitrile/water system.