US 2016/0340704 A1 · METHOD OF MAKING A BENZYLISOQUINOLINE ALKALOID (BIA) METABOLITE, ENZYMES THEREFORE

Abstract

There is provided a method of preparing a benzylisoquinoline alkaloid (BIA) metabolite comprising: a. culturing a host cell under conditions suitable for protein production, including a pH of between about 7 and about 10 said host cell comprising: b. a first heterologous coding sequence encoding a first enzyme involved in a metabolite pathway that converts (R,S)-norlaudanosoline into the metabolite; c. a second heterologous coding sequence encoding a second enzyme involved in a metabolite pathway that converts (R,S)-norlaudanosoline into the metabolite; d. a third heterologous coding sequence encoding a second enzyme involved in a metabolite pathway that converts (R,S)-norlaudanosoline into the metabolite; (d) adding (R,S)-norlaudanosoline to the cell culture; and recovering the metabolite from the cell culture

Claims111

Dependent claims

A method of preparing a benzylisoquinoline alkaloid (BIA) metabolite comprising:

(a) culturing a host cell under conditions suitable for protein production, including a first fermentation at a pH of between about 7 and about 10, and, optionally followed by a second fermentation at a pH between about 3 and about 6, said host cell comprising:
a. a first heterologous coding sequence encoding a first enzyme involved in a metabolite pathway that converts (R,S)-norlaudanosoline into the metabolite;
b. a second heterologous coding sequence encoding a second enzyme involved in a metabolite pathway that converts (R,S)-norlaudanosoline into the metabolite;
and
c. a third heterologous coding sequence encoding a third enzyme involved in a metabolite pathway that converts (R,S)-norlaudanosoline into the metabolite;
(b) adding a substrate that is (R,S)-norlaudanosoline, (R,S)-reticuline or (S)-stylopine, to the cell culture;
and
(c) recovering the metabolite from the cell culture.

5 more elements

Dependencies are inferred from the claim text.

Method of making a benzylisoquinoline alkaloid (bia) metabolite, enzymes therefore

Life of this patent

Method of making a benzylisoquinoline alkaloid (bia) metabolite, enzymes therefore

Life of this patent

Abstract

Claims111