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AssigneeDupont Nutrition Biosciences Aps
InventorsJanne Brunstedt, Jens Frisbæk Sorensen, Jørn Borch Søe, Charlotte Johansen Vedel, Birgitte Ø. Wittschieben, Jørn Dalgaard Mikkelsen, Charlotte Horsmans Poulsen, Lene B. Jensen, Lone B. Miller, Morten K. Larsen, Rikke H. Lorentzen, Charlotte R. Thoudahl, Marguerite A. Cervin, Richard R. Bott
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The present invention provides a method for preparing a variant lipid acyltransferase enzyme by expressing a nucleotide sequence encoding a lipid acyltransferase which may comprise at least one modification at a position(s) which corresponds in the encoded amino acid sequence to an amino acid(s) located in a) the canyon region of the enzyme (i.e. preferably amino acid residues 31, 27, 85, 86, 119, and 120); and/or b) insertion site 1 (i.e. amino acid residues 22-36) and/or c) insertion site 2 (i.e. amino acid residues 74-88), wherein the canyon region, insertion site 1 and/or insertion site 2 are defined as that region which when aligned based on primary or tertiary structure corresponds to the canyon region, insertion site 1 or insertion site 2 (or the corresponding amino acid residues taught above) of the enzyme shown herein as SEQ ID No. 16 or 6 in a host organism.
An isolated variant lipid acyltransferase polypeptide obtained by the method comprising expressing in a host organism a nucleotide sequence encoding a lipid acyltransferase which has at least 80% identity with the amino acid sequence of SEQ ID No. 6 and comprises at least one modification at a position(s) which corresponds in the encoded amino acid sequence to an amino acid(s) located at position 27, 31, 85, 119, 120, 122, 201, 235, 232, 245 and/or 236.
An isolated variant lipid acyltransferase polypeptide encoded by a nucleotide sequence-encoding a lipid acyltransferase which has at least 80% identity with the amino acid sequence of SEQ ID No. 6 and comprises at least one modification at a position(s) which corresponds in the encoded amino acid sequence to an amino acid(s) located at position 27, 31, 85, 119, 120, 122, 201, 235, 232, 245 and/or 236;
An isolated polypeptide which has lipid acyltransferase activity and comprises an amino acid sequence which is at least 80% identical with the amino acid sequence of SEQ ID No. 6 and comprises one or more modifications at one or more of the following positions:
An isolated variant lipid acyltransferase polypeptide comprising an amino acid sequence that is at least 70% identical to the lipid acyltransferase from Aeromonas salmonicida comprising the amino acid sequence of SEQ ID No. 6 or 16, wherein a substrate chain length specificity determining segment that lies immediately N-terminal of the Asp residue of the catalytic triad of said altered lipid acyltransferase has an altered length relative to said lipid acyltransferase from Aeromonas salmonicida comprising the amino acid sequence of SEQ ID No. 6 or 16.
Dependencies are inferred from the claim text.