Microbial production of succinate derived products
Microbes and methods used to convert renewable carbon sources such as glucose, sucrose, biomass hydrolysate, methanol or formate, to succinate-derived products, such as fumarate or malate, which are desirable products having many uses.
1. An engineered bacteria having
a deletion of one or more fumarase enzymes and able to convert succinate to fumarate; and
an overexpressed succinyl co-A ligase and succinical-semialdehyde dehydrogenase and succinic semialdehyde reductase (EC 1.1.1.B47) and able to convert succinate to 4-hydroxybutyrate.
2. The bacteria of claim 1 , further comprising a) overexpressed pyruvate carboxylase and b) deletions of i) ldhA, ii) adhE, iii) iclR and iv) ackA or pta or ackA-pta.
3. A method of making a succinate-derived product, comprising:
a. growing an engineered bacteria of claim 1 in a culture medium;
b. allowing said bacteria to anaerobically make succinate;
c. allowing said bacteria to convert succinate to a succinate-derived product under aerobic conditions; and
d. isolating said succinate-derived product from said bacteria, said medium, or both.
4. The method of claim 3 , wherein said bacteria has overexpressed pyruvate carboxylase.
5. The method of claim 4 , wherein said bacteria has a deletion of i) ldhA, ii) adhE, iii) iclR and iv) ackA or pta or ackA-pta.
6. The method of claim 3 , wherein said bacteria has a deletion of i) ldhA, ii) adhE, iii) iclR and iv) ackA or pta or ackA-pta.
7. The method of claim 3 , wherein said bacteria comprises:
a. overexpressed pyruvate carboxylase;
b. a deletion of i) ldhA, ii) adhE, iii) iclR and iv) ackA or pta or ackA-pta; and
c. said bacteria produces at least 1.2 moles of fumarate per mole of glucose.