Recombinant plants and microorganisms having a reverse glyoxylate shunt
AssigneeThe Regents of the University of California
InventorsJames C. Liao, Luisa Gronenberg, Samuel Mainguet, Sio Si Wong, Avinash Srivastava
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Abstract
Provided are microorganisms and plants that express or overexpress enzymes that catalyze the conversion of a four carbon metabolite (malate) to acetyl-CoA. Also provided are methods of generating such organisms and plants and methods of synthesizing biomass, biofuel, oil, chemicals and biochemicals using such organisms and plants.
Claims38
A recombinant microorganism comprising a metabolic pathway for the synthesis of acetyl-CoA and isocitrate from a four-carbon substrate using a pathway comprising one or more polypeptides having malate thiokinase activity, malyl-CoA lyase activity and/or isocitrate lyase activity.
3-6. (canceled)
19-20. (canceled)
(canceled)
A cell-free system for converting a 4-carbon substrate to isocitrate and two acetyl-CoAs comprising ATP and CoA and:
- (i) an enzyme the converts malate to malyl-CoA;
- (ii) an enzyme the converts malyl-CoA to glycosylate and acetyl-CoA;
- (iii) an enzyme that converts isocitrate to citrate;
- and
- (iv) an enzyme that converts citrate to oxaloacetate.
A recombinant microorganism for producing 1-butanol, wherein the microorganism comprises:
- (i) an enzyme the converts malate to malyl-CoA;
- (ii) an enzyme the converts malyl-CoA to glycosylate and acetyl-CoA;
- (iii) an enzyme that converts isocitrate to citrate;
- (iv) an enzyme that converts citrate to oxaloacetate;
- (v) an enzyme that converts acetyl-CoA to acetoacetyl-CoA, and at least one enzyme that converts (a) acetoacetyl-CoA to (R)- or (S)-3-hydroxybutyryl-CoA and (R)- or (S)-3-hydroxybutyryl-CoA to crotonyl-CoA,
- (vi) an enzyme that converts crotonyl-CoA to butyryl-CoA;
- and
- (vii) an enzyme that converts butyryl-CoA to butyraldehyde and butyraldehyde to 1-butanol.
A recombinant plant engineered to express one or more polypeptides having activity selected form the group consisting of malate thiokinase activity, malyl-CoA lyase activity, pyruvate:ferrodoxin oxidoreductase activity and fumarase reductase activity and wherein the recombinant plant produces more acetyl-CoA compared to a wild-type of parental plant.
36-41. (canceled)
45-49. (canceled)
52-67. (canceled)
Dependencies are inferred from the claim text.