caa a22 4500 465822150 CHVBK 20180323112138.0 cr unu---uuuuu 170327e19900301xx s 000 0 eng 10.1007/BF00164734 doi (NATIONALLICENCE)springer-10.1007/BF00164734 Subtilisin-catalysed peptide synthesis and transesterification in organic solvents [Elektronische Daten] [Anne Ferjancic, Antoine Puigserver, Hubert Gaertner] Summary: Subtilisin from Bacillus subtilis was modified with polyethylene glycol (PEG), or adsorbed either on celite or porous glass, or directly used as a suspended powder to catalyse peptide synthesis and transesterification reactions in organic solvents. The rather low yield of peptide synthesis probably resulted from the enzyme tendency to catalyse hydrolysis and transesterification side reactions. The kinetics of transesterification catalysed by PEG-subtilisin was consistent with a ping-pong mechanism modified by a hydrolytic branch. Initial rates of transesterification were found to be dependent on alcohol and organic base concentrations in the reaction mixture. The high affinity of benzyloxycarbonyl-l-serine-methyl ester for the enzyme indicated that a change in substrate specificity of subtilisin occurred in organic phase. The 50-fold increase in the rate of synthesis of benzyloxycarbonyl-l-serine-l-phenylalanine amide which was observed when PEG-subtilisin was used instead of immobilized or powdered enzyme, suggested that a higher flexibility of the polypeptide chain modified by the covalent attachment of a number of soluble PEG moieties occurred in organic solvents. This also resulted in a lower stability of PEG-subtilisin at high temperature. Springer-Verlag, 1990 Ferjancic Anne Centre de Biochimie et de Biologie Moléculaire, Centre National de la Recherche Scientifique, BP 71, F-13402, Marseille Cedex 9, France aut Puigserver Antoine Centre de Biochimie et de Biologie Moléculaire, Centre National de la Recherche Scientifique, BP 71, F-13402, Marseille Cedex 9, France aut Gaertner Hubert Centre de Biochimie et de Biologie Moléculaire, Centre National de la Recherche Scientifique, BP 71, F-13402, Marseille Cedex 9, France aut Applied Microbiology and Biotechnology Springer-Verlag 32/6(1990-03-01), 651-657 0175-7598 32:6<651 1990 32 253 https://doi.org/10.1007/BF00164734 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF00164734 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Ferjancic Anne Centre de Biochimie et de Biologie Moléculaire, Centre National de la Recherche Scientifique, BP 71, F-13402, Marseille Cedex 9, France aut NATIONALLICENCE 700 1- Puigserver Antoine Centre de Biochimie et de Biologie Moléculaire, Centre National de la Recherche Scientifique, BP 71, F-13402, Marseille Cedex 9, France aut NATIONALLICENCE 700 1- Gaertner Hubert Centre de Biochimie et de Biologie Moléculaire, Centre National de la Recherche Scientifique, BP 71, F-13402, Marseille Cedex 9, France aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer-Verlag 32/6(1990-03-01), 651-657 0175-7598 32:6<651 1990 32 253 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer