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AssigneeArcher Daniels Midland Company
InventorsOwen Ryan, Alexander Chou, Jesús Seco Moral, Marina Canellas, Jamie Ryding
Estimating the term…
Described herein is a novel enzymatic pathway and acid tolerant Schizosaccharomyces pombe strain exhibiting the same, for producing 3-hydroxypropionic acid by fermentation wherein the S. pombe strain includes nucleotide sequences that encode, and which are operably linked to promoters to express, non-native enzymes that exhibit an oxaloacetate decarboxylase (ODC) activity that converts oxaloacetate to 3-oxopropionate and a 3-oxopropionate reductase (OPR) activity that converts the 3-opropionate to 3-hydroxypropianate. ODC and OPR enzymes are not known to exist in nature, nor is an enzymatic path for making 3-hydroxypropionic acid but particular enzymes exhibiting the requisite activities are herein identified by sequence. A further enhancement is to also overexpress an enzyme that has at least one of a pyruvate carboxylase activity, a phosphoenolpyruvate carboxy kinase activity, and a phosphoenolpyruvate carboxylase activity, which increases the level of oxaloacetate in the cell leading to greater 3-HP production.
An acid tolerant yeast strain comprising nucleotide sequences that include a promoter sequence operable in the microorganism operably linked to coding sequences that encode non-native enzymes that exhibit a) an oxaloacetate decarboxylase (ODC) activity that converts oxaloacetate to 3-oxopropionate;
An acid tolerant yeast strain comprising a nucleotide sequences that include a promoter sequence operable in the microorganism operably linked to coding sequences that encode non-native enzymes that exhibit a) an oxaloacetate decarboxylase (ODC) activity that converts oxaloacetate to 3-oxopropionate, wherein the ODC activity is provided by an enzyme having a sequence selected from the group consisting of SEQ ID NOS:
A method of determining whether a candidate enzyme increases oxaloacetate production in a microorganism comprising, expressing the candidate enzyme in the microorganism while simultaneously expressing an exogenous malate dehydrogenase gene in the microorganism and measuring the production of malic acid in the microorganism wherein an increase of malic acid production is determinative of whether the candidate enzyme increases oxaloacetate production in the microorganism.
Dependencies are inferred from the claim text.