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.
Claims6
A Thermotoga maritima strain comprising at least one mutation in the ATP-binding component of a maltose ABC transporter, wherein the ATP-binding component of the maltose ABC transporter comprises the amino acid sequence of SEQ ID NO:15 and at least one mutation selected from V233S or V233F, wherein the T. maritima strain overproduces molecular hydrogen (H2) compared to a T. maritima strain lacking the at least one mutation.
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 and isolating the compensating T. maritima having an increase in H2, wherein the compensating mutation is in the ATP-binding component of a maltose ABC transporter, wherein the ATP-binding component of the maltose ABC transporter comprises the amino acid sequence of SEQ ID NO:15 and at least one mutation selected from V233S or V233F.
Dependencies are inferred from the claim text.