公开(公告)号：US10927399B2

公开(公告)日：2021-02-23

申请号：US14045683

申请日：2013-10-03

Applicant: DSM IP ASSETS B.V.

Inventor： Ulrike Maria Mueller ,  Liang Wu ,  Lourina Madeleine Raamsdonk ,  Aaron Adriaan Winkler

IPC: C12N1/16 ,  C12N9/04 ,  C12Q1/32 ,  C07K14/33 ,  C12N9/02 ,  C12P7/16

Abstract: The present invention relates to a method of identifying a heterologous polypeptide having enzymatic activity for converting pyruvate, acetaldehyde or acetate into acetyl-CoA in (the cytosol of) a yeast cell comprising: a) providing a mutated yeast cell comprising a deletion of at least one gene of the (PDH) by-pass, selected from the genes encoding the enzymes pyruvate decarboxylase (PDC), acetaldehyde dehydrogenase (ALD), and acetyl-CoA synthetase (ACS); b) transforming said mutated yeast cell with an expression vector comprising a heterologous nucleotide sequence encoding a candidate polypeptide having potential enzymatic activity for converting pyruvate, acetaldehyde or acetate into acetyl-CoA; c) testing said recombinant mutated yeast cell for its ability to grow on minimal medium containing glucose as sole carbon source, and d) identifying said candidate polypeptide as a heterologous polypeptide having enzymatic activity for converting pyruvate, acetaldehyde or acetate into acetyl-CoA in (the cytosol of) said yeast cell when growth of said cell is observed. The invention further relates to a method of producing a fermentation production such as butanol.