Metabolic engineering of arabinose-fermenting yeast cells

摘要

The invention relates to an eukaryotic cell expressing nucleotide sequences encoding the ara A, ara B and ara D enzymes whereby the expression of these nucleotide sequences confers on the cell the ability to use L-arabinose and/or convert L-arabinose into L-ribulose, and/or xylulose 5-phosphate and/or into a desired fermentation product such as ethanol. Optionally, the eukaryotic cell is also able to convert xylose into ethanol.

主权项

1. A eukaryotic cell capable of expressing the following nucleotide sequences, wherein the expression of these nucleotide sequences confers on the cell the ability to use L-arabinose and/or to convert L-arabinose into L-ribulose, and/or xylulose 5-phosphate and/or into a desired fermentation product:
(a) a nucleotide sequence encoding an arabinose isomerase (araA), wherein said nucleotide sequence is selected from the group consisting of:
i. nucleotide sequences encoding an araA, said araA comprising an amino acid sequence that has at least 90% sequence identity with the amino acid sequence of SEQ ID NO:1,ii. nucleotide sequences comprising a nucleotide sequence that has at least 90% sequence identity with the nucleotide sequence of SEQ ID NO:2,iii. nucleotide sequences the complementary strand of which hybridizes to a nucleic acid molecule of sequence of (i) or (ii) under moderate conditions; (b) a nucleotide sequence encoding a L-ribulokinase (araB), wherein said nucleotide sequence is selected from the group consisting of:
i. nucleotide sequences encoding an araB, said araB comprising an amino acid sequence that has at least 90% sequence identity with the amino acid sequence of SEQ ID NO:3,ii. nucleotide sequences comprising a nucleotide sequence that has at least 90% sequence identity with the nucleotide sequence of SEQ ID NO:4,iii. nucleotide sequences the complementary strand of which hybridizes to a nucleic acid molecule of sequence of (i) or (ii) under moderate conditions; (c) a nucleotide sequence encoding an L-ribulose-5-P-4-epimerase (araD), wherein said nucleotide sequence is selected from the group consisting of:
i. nucleotide sequences encoding an araD, said araD comprising an amino acid sequence that has at least 90% sequence identity with the amino acid sequence of SEQ ID NO:5,ii. nucleotide sequences comprising a nucleotide sequence that has at least 90% sequence identity with the nucleotide sequence of SEQ ID NO:6,iii. nucleotide sequences the complementary strand of which hybridizes to a nucleic acid molecule of sequence of (i) or (ii) under moderate conditions; wherein moderate conditions is defined as conditions that allow a nucleic acid sequences of at least 50 nucleotides to hybridize at a temperature of about 45° C. in a solution comprising about 1 M salt and washing at room temperature in a solution comprising about 1 M salt.