Mutant microorganisms and methods of making and using
AssigneeNUtech Ventures
InventorsRaghuveer Singh, Derrick White, Paul Blum
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Abstract
This disclosure describes methods that allow for the uncoupling of microbial growth from product formation, which allows for maximal use of raw material and optimal end-product formation.
Claims23
A method of making a mutant microorganism that produces an increased amount of a biologically-produced metabolite relative to a microorganism lacking the mutation(s), the method comprising:
- disrupting, transiently, a gene in a microorganism, thereby producing a transiently disrupted microorganism;
- passaging the transiently disrupted microorganism a plurality of times under selective pressure to produce a compensating microorganism;
- screening the compensating microorganism for an increase in one or more metabolites;
- and
- isolating the compensating microorganism having an increase in the desired metabolite.
A Thermotoga maritima strain comprising at least one mutation, wherein the T. maritima strain overproduces molecular hydrogen (H2).
A method of increasing the yield of molecular hydrogen (H2) produced by Thermotoga maritima in culture, comprising:
- inactivating, transiently, the lactate dehydrogenase gene in the T. maritima using a selectable marker to produce an inactivated T. maritima;
- passaging the inactivated T. maritima a plurality of times under selective pressure to produce a compensating T. maritima, wherein the compensating T. maritima comprises a compensating mutation;
- screening the compensating T. maritima for an increase in H2;
- and
- selecting/isolating the compensating T. maritima having an increase in H2.
Dependencies are inferred from the claim text.