Microbiological production method for...

C - Chemistry – Metallurgy – 12 – N

Patent

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C12N 15/52 (2006.01) C07K 5/06 (2006.01) C12N 1/21 (2006.01) C12N 9/00 (2006.01) C12N 15/09 (2006.01) C12N 15/62 (2006.01) C12P 21/02 (2006.01)

Patent

CA 2365594

The present invention relates to a new, microbiological method for the production of .alpha.-L-aspartyl-L-phenylalanine (Asp-Phe) from the substrates L-aspartic acid (L-Asp) and L-phenylalanine (L-Phe) wherein the substrates are contacted, in the presence of ATP, with a non-ribosomal dipeptide synthetase comprising two minimal modules connected by one condensation domain wherein the N- resp. C-terminal modules are recognising L-Asp and L-Phe, respectively, and the latter module is covalently bound at its N- terminal end to the condensation domain, and wherein each of these minimal modules is composed of an adenylation domain and a 4'-phosphopantetheinyl cofactor containing thiolation domain, and that the Asp-Phe formed is recovered. The present invention also relates to novel DNA fragments or combination of DNA fragments encoding a new Asp-Phe dipeptide synthetase, micro-organisms containing such DNA fragments, as well as to the new Asp-Phe dipeptide synthetases itself.

La présente invention concerne une nouvelle méthode microbiologique de production d'.alpha.-L-aspartyl-L-phénylalanine (Asp-Phe) à partir des substrats acide L-aspartique (L-Asp) et L-phénylalanine (L-Phe) dans laquelle les substrats sont mis en contact, en la présence d'ATP, avec une synthétase dipeptidique non ribosomique contenant deux modules minimaux reliés par un domaine de condensation dans lequel les modules C-terminaux N-resp reconnaissent L-Asp et L-Phe respectivement, et le dernier module est lié de manière covalente au niveau de son extrémité N-terminale au domaine de condensation, chacun de ces modules minimaux étant composés d'un domaine d'adénylation et d'un domaine de thiolation contenant un cofacteur 4'-phosphopantétheinyle, et le Asp-Phe formé est récupéré. La présente invention concerne également de nouveaux fragments d'ADN ou une combinaison de fragemts d'ADN codant une nouvelle synthétase dipeptidique Asp-Phe, des micro-organismes contenant ces fragments d'ADN, ainsi que les nouvelles synthétases dipeptidiques Asp-Phe elles-mêmes.

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