Enzymatic synthesis of a CCK-8 tripeptide fragment in organic media

M. Capellas, M. D. Benaiges, G. Caminal, G. Gonzalez, J. Lopez-Santín, P. Clapés

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    34 Citations (Scopus)

    Abstract

    The enzymatic synthesis of the tripeptide derivative Z-Gly-Trp-Met-OEt is reported. This tripeptide is a fragment of the cholecystokinin C-terminal octapeptide CCK-8. Studies on the α-chymotrypsin catalyzed coupling reaction between Z-Gly-Trp-R1 and Met-R2 have focused on low water content media, using deposited enzyme on inert supports such as Celite and polyamide. The effect of additives (polar organic solvents), the acyl-donor ester structure, the C-α protecting group of the nucleophile, enzyme loading, and substrate concentration were tested. The best reaction medium found was acetonitrile containing buffer (0.5%, v/v) and triethylamine (0.5%, v/v) using the enzyme deposited on Celite as catalyst (8 mg of α-chymotrypsin/g of Celite). A reaction yield of 81% was obtained with Z-Gly-Trp-OCam as acyl donor, at an initial concentration of 80 mM. The tripeptide synthesis was scaled up to the production of 2 g of pure tripeptide with an overall yield of 71%, including reaction and purification steps.
    Original languageEnglish
    Pages (from-to)700-708
    JournalBiotechnol. Bioeng.
    Volume50
    Issue number6
    DOIs
    Publication statusPublished - 20 Jun 1996

    Keywords

    • α-chymotrypsin
    • CCK-8
    • organic media
    • tripeptide synthesis

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