TY - JOUR

T1 - Monolithic bioreactors

T2 - Effect of chymotrypsin immobilization on its biocatalytic properties

AU - Ponomareva, E. A.

AU - Kartuzova, V. E.

AU - Vlakh, E. G.

AU - Tennikova, T. B.

N1 - Funding Information: The work was supported by Russian Fund of Basic Researches (grant RFBR #08-08-00876-a ), State Support of Leading Scientific Schools (grant #4391.2008.3 ) and by personal financing of Dr. E. Vlakh by Russian Science Support Foundation (RSSF, 2009). The authors are grateful personally to Dr. Aleš Štrancar and BIA Separations d.o.o. (Ljublajana, Slovenia) for kindly providing CIM monolithic minidisks, as well as to Drs. N.D. Ivanova, G.A. Platonova and I.V. Kalashnikova (all from IMC RAS) for fruitful discussions.

PY - 2010/2/15

Y1 - 2010/2/15

N2 - The effect of different modes of α-chymotrypsin attachment to the surface of methacrylate-based ultrashort monolithic minicolumns on enzyme activity has been studied. The immobilization of protease was carried out via direct covalent binding of chymotrypsin, as well as via its attachment through small and polymer spacers. It was established that the lowest enzyme activity against N-benzoyl-l-tyrosine ethyl ester was found for bioreactor obtained via direct attachment of chymotrypsin to the surface of GMA-EDMA minidisks, whereas the highest parameter close to that determined for dissolved enzyme was found in the case of bioreactor prepared by the introduction of copolymer of 2-deoxy-N-methacryloylamido-d-glucose with N-vinylpyrrolidone and acrolein as a long and flexible polymer spacer. Additionally, the effect of flow rate of substrate recirculation on bioconversion efficiency was examined. Independently on immobilization method, the increase of flow rate led to the raise of biocatalytic efficiency.

AB - The effect of different modes of α-chymotrypsin attachment to the surface of methacrylate-based ultrashort monolithic minicolumns on enzyme activity has been studied. The immobilization of protease was carried out via direct covalent binding of chymotrypsin, as well as via its attachment through small and polymer spacers. It was established that the lowest enzyme activity against N-benzoyl-l-tyrosine ethyl ester was found for bioreactor obtained via direct attachment of chymotrypsin to the surface of GMA-EDMA minidisks, whereas the highest parameter close to that determined for dissolved enzyme was found in the case of bioreactor prepared by the introduction of copolymer of 2-deoxy-N-methacryloylamido-d-glucose with N-vinylpyrrolidone and acrolein as a long and flexible polymer spacer. Additionally, the effect of flow rate of substrate recirculation on bioconversion efficiency was examined. Independently on immobilization method, the increase of flow rate led to the raise of biocatalytic efficiency.

KW - Enzyme immobilization

KW - Polymer monolithic sorbents

KW - Polymer spacer

UR - http://www.scopus.com/inward/record.url?scp=75349107235&partnerID=8YFLogxK

U2 - 10.1016/j.jchromb.2010.01.001

DO - 10.1016/j.jchromb.2010.01.001

M3 - Article

C2 - 20106728

AN - SCOPUS:75349107235

VL - 878

SP - 567

EP - 574

JO - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences

JF - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences

SN - 1570-0232

IS - 5-6

ER -