14-02-2019 дата публикации
Номер: US20190048375A1
Принадлежит:
The present invention relates to processes to make neosaxitoxin, and analogues and variants thereof, and intermediates in the production of neosaxitoxin in recombinant host cells. Neosaxitoxin and the analogues and variants thereof may be used in the production of pharmaceutical compositions. 1. A process for producing neosaxitoxin in a host cell , the process comprising the steps: (i) S-adenosylmethionine,', '(ii) arginine', '(iii) acetyl-CoA, malony-CoA or propionyl-CoA, and', '(iv) carbamoyl phosphate,, '(A) culturing a host cell which comprises nucleic acid molecules encoding a PPTase and encoding the Sxt polypeptides A, B, D, G, H, I, S, T, U, V, W and X in a culture medium in the presence of the substratesand wherein the host cell does not comprise nucleic acid molecules encoding the Sxt polypeptides C, F, J, K, L, M, P, Q, R and ORF24,under conditions which are suitable for the production of neosaxitoxin; and optionally(B) isolating and/or purifying neosaxitoxin from the host cells or from the culture medium.2. A process for producing neosaxitoxin or an analogue or variant thereof , the process comprising the steps: (i) S-adenosylmethionine,', '(ii) arginine', '(iii) acetyl-CoA, malony-CoA or propionyl-CoA, and', '(iv) carbamoyl phosphate,, '(A) contacting the substrateswith Sxt A, B, D, G, H, I, S, T, U, V, W and X polypeptides, in a reaction medium, and optionally(B) isolating and/or purifying neosaxitoxin or an analogue or variant thereof from the reaction medium.3. A process as claimed in claim 2 , wherein the reaction medium additionally comprises a PPTase.4. A process for producing neosaxitoxin or an analogue or variant thereof in a host cell claim 2 , the process comprising the steps: (i) S-adenosylmethionine,', '(ii) arginine', '(iii) acetyl-CoA, malony-CoA or propionyl-CoA, and', '(iv) carbamoyl phosphate,, '(A) culturing a host cell which comprises nucleic acid molecules encoding the Sxt polypeptides A, B, D, G, H, I, S, T, U, V, W and X in a ...
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