Phosphoketolases for improved production of acetyl coenzyme A-derived metabolites, isoprene, isoprenoid precursors, and isoprenoids
AssigneesDanisco US Inc., The Goodyear Tire & Rubber Company
InventorsZachary Q. Beck, Jeffrey W. Munos, Derek H. Wells, Jian Yao
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Abstract
This present invention relates to cultured recombinant cells comprising a heterologous phosphoketolase (PKL) polypeptide that are capable of increased production of acetyl coenzyme A-derived metabolites, as well as methods for producing and using the same. In some embodiments, the recombinant cells further comprise one or more mevalonate (MVA) pathway polypeptides for the production of isoprenoid precursors, isoprene and isoprenoids.
Claims20
A recombinant cell capable of increased carbon flux through the phosphoketolase pathway, wherein the recombinant cell comprises:
- (i) a heterologous nucleic acid sequence encoding a polypeptide having phosphoketolase activity, wherein the polypeptide comprises at least 65% sequence identity to SEQ ID NO:8 or SEQ ID NO:11 and (ii) one or more nucleic acids encoding one or more polypeptides of the complete mevalonate (MVA) pathway, wherein the recombinant cell is a recombinant Escherichia coli cell, and wherein
- (1) said recombinant cell comprising said polypeptide having phosphoketolase activity of (i) has a Performance Index value of greater than 1.0 in one or more of the following parameters:
- (a) cell growth on glucose, (b) cell growth on xylose, (c) production of intracellular acetyl-phosphate or (d) cell growth on glucose-6-phosphate, wherein the Performance Index value is calculated as activity per unit relative to a corresponding cell expressing a phosphoketolase from E. gallinarum or
- (2) said polypeptide having phosphoketolase activity of (i) has a Performance Index value of greater than 1.0 in one or more of the following parameters:
- (e) protein solubility, (f) protein expression, or (g) fructose-6-phosphate (F6P) Specific Activity, wherein the Performance Index value is calculated as activity per unit relative to a phosphoketolase from E. gallinarum.
Dependencies are inferred from the claim text.