Recombinant host cells comprising phosphoketolases
AssigneeButamax Advanced Biofuels LLC
InventorsMichael Dauner, Lori Ann Maggio-Hall, Jean-Francois Tomb
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Abstract
The present invention is related to recombinant host cells comprising: (i) at least one deletion, mutation, and/or substitution in an endogenous gene encoding a polypeptide that converts pyruvate to acetaldehyde, acetyl-phosphate or acetyl-CoA; and (ii) a heterologous polynucleotide encoding a polypeptide having phosphoketolase activity. The present invention is also related to recombinant host cells further comprising (iii) a heterologous polynucleotide encoding a polypeptide having phosphotransacetylase activity.
Claims48
A recombinant host cell comprising:
- (i) at least one genetic modification in an endogenous gene encoding a polypeptide having pyruvate decarboxylase activity, wherein the at least one genetic modification reduces or eliminates pyruvate decarboxylase activity;
- (ii) a heterologous polynucleotide encoding a polypeptide having phosphoketolase activity;
- and
- (iii) a pyruvate-utilizing biosynthetic pathway, wherein said pyruvate-utilizing biosynthetic pathway is an isobutanol biosynthetic pathway comprising the substrate to product conversions:
- (a) pyruvate to acetolactate;
- (b) acetolactate to 2,3-dihydroxyisovalerate;
- (c) 2,3-dihydroxyisovalerate to 2-ketoisovalerate;
- (d) 2-ketoisovalerate to isobutyraldehyde;
- and
- (e) isobutyraldehyde to isobutanol;
- and
- wherein said recombinant host cell produces isobutanol.
A recombinant host cell comprising:
- (i) at least one genetic modification in an endogenous gene encoding a polypeptide having pyruvate decarboxylase activity, wherein the at least one genetic modification reduces or eliminates pyruvate decarboxylase activity;
- (ii) a heterologous polynucleotide encoding a polypeptide having phosphoketolase activity;
- and
- (iii) a pyruvate-utilizing biosynthetic pathway, wherein said pyruvate-utilizing biosynthetic pathway is a 2-butanone biosynthetic pathway comprising the substrate to product conversions:
- (i) pyruvate to acetolactate;
- (ii) acetolactate to acetoin;
- (iii) acetoin to 2,3-butanediol;
- (iv) 2,3-butanediol to 2-butanone;
- and
- wherein said recombinant host cell produces 2-butanone.
A recombinant host cell comprising:
- (i) at least one genetic modification in an endogenous gene encoding a polypeptide having pyruvate decarboxylase activity, wherein the at least one genetic modification reduces or eliminates pyruvate decarboxylase activity;
- (ii) a heterologous polynucleotide encoding a polypeptide having phosphoketolase activity;
- and
- (iii) a pyruvate-utilizing biosynthetic pathway, wherein said pyruvate-utilizing biosynthetic pathway is a 2-butanol biosynthetic pathway comprising the substrate to product conversions:
- (i) pyruvate to acetolactate;
- (ii) acetolactate to acetoin;
- (iii) acetoin to 2,3-butanediol;
- (iv) 2,3-butanediol to 2-butanone;
- (v) 2-butanone to 2-butanol;
- and
- wherein said recombinant host cell produces 2-butanol.
A method of improving the growth of a recombinant host cell comprising at least one genetic modification in an endogenous gene encoding a polypeptide having pyruvate decarboxylase activity, wherein the at least one genetic modification reduces or eliminates pyruvate decarboxylase activity, comprising:
- transforming said recombinant host cell with a heterologous polynucleotide encoding a polypeptide having phosphoketolase activity,
- wherein the recombinant host cell comprising the heterologous polynucleotide encoding a polypeptide having phosphoketolase activity has improved growth as compared to a control recombinant host cell without the heterologous polynucleotide encoding a polypeptide having phosphoketolase activity.
A method of reducing the requirement for an exogenous two-carbon substrate for the growth of a recombinant host cell comprising at least one genetic modification in an endogenous gene encoding a polypeptide having pyruvate decarboxylase activity, wherein the at least one genetic modification reduces or eliminates pyruvate decarboxylase activity, comprising:
- transforming said recombinant host cell with a heterologous polynucleotide encoding a polypeptide having phosphoketolase activity, wherein the recombinant host cell comprising the heterologous polynucleotide encoding a polypeptide having phosphoketolase activity has a reduced requirement for an exogenous two-carbon substrate as compared to a control recombinant host cell without the heterologous polynucleotide encoding a polypeptide having phosphoketolase activity.
Dependencies are inferred from the claim text.