Methionine analogue synthesis
AssigneeenGenes Biotech GmbH
InventorsBirgit Wiltschi, Patrik Fladlscher, Lisa-Ramona Offner-Marko, Kathrin Heckenbichier, Rolf Breinbauer
Life of this patent
Estimating the term…
Abstract
The present invention relates to a method for preparing a methionine analogue, comprising contacting a host cell with L-homoserine and one or more nucleophiles and cultivating said host cell and said one or more nucleophiles of (a) in a fermentation broth to produce said methionine analogue, wherein said host cell encodes and stably expresses a L-homoserine-O-acetyl-transferase (HSAT) and an O-acetyl-L-homoserine sulfhydrylase (OAHS). The present invention also relates to host cells encoding and expressing such HSAT and OAHS, as well as uses thereof for producing a methionine analogue.
Claims20
Method for preparing a methionine analogue, comprising the following steps:
- (a) contacting a host cell with L-homoserine and one or more nucleophiles;
- and
- (b) cultivating said host cell and said one or more nucleophiles of (a) in a fermentation broth to produce said methionine analogue,
- wherein said host cell encodes and stably expresses a L-homoserine-O-acetyl-transferase (HSAT) and an O-acetyl-L-homoserine sulfhydrylase (OAHS), wherein said O-acetyl-L-homoserine sulfhydrylase (OAHS) has an amino acid sequence having up to 50 amino acid deviations compared to the amino acid sequence of SEQ ID NO:
- 1, and said OAHS is able to convert O-acetyl-L-homoserine to a methionine analogue.
Method of any one of the preceding claims, wherein said methionine analogue is an amino acid comprising an azido, alkyne, alkene, keto, aldehyde, norbornene, or furan moiety in its side chain.
Method of any one of the preceding claims, wherein said methionine analogue is selected from the group consisting of S-allyl-L-homocysteine ((2S)-2-amino-4-(prop-2-en-1-ylsulfanyl)butanoic acid;
- Sen or Sahc), S-propargyl-L-homocysteine (Syn), S-furyl-L-homocysteine (Fur), S-furfuryl-L-homocysteine (SFur), O-furfuryl-L-homoserine (OFur), O-allyl-L-homoserine (Oen), O-propargyl-L-homoserine (O-propargyloxy-L-homoserine, Oyn), L-azidohomoalanine (Aha), S-azidopropyl-L-homocysteine (Saz or SN3), O-norbornenemethoxy-L-homoserine ((2S)-2-amino-4-(bicyclo[2.2.1]hept-5-en-2-ylmethoxy)butanoic acid;
- Nom), O-norbornenoxy-L-homoserine ((2S)-2-amino-4-(bicyclo[2.2.1]hept-5-en-2-yloxy)butanoic acid;
- Nor), O-[2-(1,2,5-dithiazepan-5-yl)-2-oxoethoxy]-L-homoserine ((2S)-2-amino-4-[2-(1,2,5-dithiazepan-5-yl)-2-oxoethoxy]butanoic acid, SEA-Hse), O-(2-oxopropoxy)-L-homoserine ((2S)-2-amino-4-(2-oxopropoxy)butanoic acid;
- Opo), and O-(3-oxobutoxy)-L-homoserine ((2S)-2-amino-4-(3-oxobutoxy)butanoic acid;
- Obo).
Method of any one of the preceding claims, wherein said one or more nucleophiles are selected from the group consisting of thiols, thioesters, thioacetates, selenols, selenoacetates, norbornene, alcohols, azides.
Method of any one of the preceding claims, wherein said L-homoserine-O-acetyl-transferase (HSAT) has an amino acid sequence having up to 50 amino acid deviations compared to the amino acid sequence of SEQ ID NO:
- 2 or 3, and said HSAT is able to convert homoserine to O-acetyl-L-homoserine.
Method of any one of the preceding claims, wherein in a further step said methionine analogue is incorporated into a protein during protein biosynthesis.
Host cell encoding and stably expressing an L-homoserine-O-acetyl-transferase (HSAT) and an O-acetyl-L-homoserine sulfhydrylase (OAHS), wherein said O-acetyl-L-homoserine sulfhydrylase (OAHS) has an amino acid sequence having up to 50 amino acid deviations compared to the amino acid sequence of SEQ ID NO:
- 1, and said OAHS is able to convert O-acetyl-L-homoserine to a methionine analogue.
Dependencies are inferred from the claim text.