Enzymatic reactions in the presence of keto acids

Details Enzymatic reactions in the presence of keto acids Enzymatic reactions in the presence of keto acids

C - Chemistry, Metallurgy

12

P

C12P 13/02 (2006.01) C12P 21/06 (2006.01)

Patent

CA 2586128

Conversion in vitro of X-Gly to X-alpha-hydroxy-Gly or X-NH2 (X being a peptide or any other compound having a carbonyl group capable of forming a covalent bond with glycine) is accomplished enzymatically in the presence of keto acids, or salts or esters thereof, to provide a good yield without the necessity of catalase or similar enzymatic reaction enhancers. Peptidylglycine .alpha.-amidating monooxygenase (PAM) is a preferred enzyme for catalyzing the conversion. Alternatively, peptidylglycine .alpha.-hydroxylating monooxygenase (PHM) is utilized to convert X-Gly to X-alpha-hdroxy-Gly which may be recovered, or optionally may be simultaneously or sequentially converted to an amid by either a Lewis base or action of the enzyme peptidyl .alpha.- hydroxyglycine A-amidating lyase (PAL). Both PHM and PAL are functional domains of PAM.

Selon l'invention, la conversion in vitro de X-Gly en X-alpha-hydroxy-Gly ou X-NH2 (X représentant un peptide ou tout autre composé à groupe carbonyle capable de former une liaison covalente avec la glycine) s'effectue au niveau enzymatique en présence d'acides cétoniques ou de sels ou d'esters associés, de manière à engendrer un meilleur rendement sans avoir recours à une catalase ou à des amplificateurs de réaction enzymatique. La peptidylglycine a-amidating monooxygénase (PAM) constitue un enzyme préféré servant à catalyser la conversion. En outre, la peptidylglycine a-hydroxylating monooxygénase (PHM) est utilisée pour convertir X-Gly en X-alpha-hydroxy-Gly qui peut être récupéré ou, facultativement, qui peut être simultanément ou séquentiellement converti en un amide soit par une base de Lewis soit par action de la peptidyl a-hydroxyglycine a-amidating lyase (PAL) enzymatique. La PHM et la PAL constituent des domaines fonctionnels de PAM.

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