Danisco A/s

Jørn Dalgaard Mikkelson

Jørn Borch Søe

Arno De Kreij

The present invention relates to a method of producing a variant lipid acyltransferase enzyme by selecting a parent enzyme which is a lipid acyltransferase enzyme having the amino acid sequence motif GDSX, modifying one or more amino acids to produce a variant lipid acyltransferase, testing the variant lipid acyltransferase for activity on a galactolipid substrate, a phospholipid substrate and/or a triglyceride substrate, selecting a variant enzyme with an enhanced activity towards galactolipids compared with the parent enzyme, and/or preparing a quantity of the variant enzyme. In some embodiments, the variant lipid acyltransferase enzyme may include the amino acid sequence motif GDSX, wherein X is one or more of the following amino acid residues L, A, V, I, F, Y, X, Q, T N, M or S, and wherein the variant enzyme has one or more amino acid modifications compared with a parent sequence.

Lipid acyltransferases

The present invention relates to a method for the production of a variant lipid acyltransferase comprising the steps of: (i) selecting a parent enzyme which is a lipid acyltranserase enzyme; (ii) modifying one or more amino acids to produce a variant lipid acyltransferase; (iii) testing the activity of the variant lipid acyltransferase on a galactolipid and/or phospholipid and/or triglyceride substrate; (iv) selecting a variant enzyme with enhanced activity towards galactolipids compared with the parent enzyme; (v) providing acell; (vi) transforming thecell with a heterologous nucleotide sequence encoding said variant lipid acyltransferase; and (iii) expressing said variant lipid acyltransferase in the cell under the control of a promoter sequence. The variant lipid acyltransferase can undergo post-translations modification, truncation and/or clipping, i.e., to remove a signal peptide. In addition, the present invention further relates to the use ofto express a lipid acyltransferase, ahost cell comprising a heterologous lipid acyltransferase and a vector comprising a nucleotide sequence encoding a lipid acyltransferase operably linked to a promoter sequence homologous to

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Hvidovre, Denmark

Jørn Dalgaard Mikkelson