Strains and Genes that Facilitate the Genetic Manipulation of Hyperthermophiles
- Detailed Technology Description
- ApplicationGeneticallymodified hyperthermophile organisms and tools for their development. Organismssuitable for the production of biofuels, hydrogen and other chemicals throughdegradation of biomass ProblemsAddressed Itwould be difficult to overestimate the contribution of genetic manipulation tothe study of any bio-logical system, and it is an essential tool for themetabolic engineering of biosynthetic and substrate utilization pathways. Thisis particularly true for the archaea since, in spite of their environmental andindustrial importance, coupled with their unique molecular features, muchremains to be learned about their biology. The marine hyperthermophilicanaerobe Pyrococcus furiosus is of special interest not only for its ability togrow optimally at 100° C and the implications of this trait for its biology butalso for industrial applications of its enzymes, as well as its capacity toproduce hydrogen efficiently in a highly efficient manner. The development ofgenetic systems in the archaea, in general, presents many unique challengesgiven the extreme growth requirements of many of these organisms. To date,genetic systems of various levels of sophistication have been developed forrepresentatives of all ma-jor groups of archaea, including halophiles,methanogens, thermoacidophiles, and hyperthermophiles. One of the mostsignificant barriers to genetic manipulation of archaea, in general, andhyperthermophiles, in particular, is the lack of selectable markers. Antibioticselection strategies used in mesophilic bacteria are typically ineffectivebecause the molecular machineries of archaea are not affected by theantibiotic.
- *Abstract
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TechnologySummary
Universityof Georgia researchers have developed methods for transforming P. furiosus withany given polynucleotide. The number of transformants may be at least103/μg(DNA) and such number can be even higher than 105/ μg(DNA). Thepolynucleotide can be circular or linear. Using this method, sever-almodifications of P. furiosus have been developed. Plasmids of the invention arestable and remain unchanged for more than 100 generations of the recipientorganism. Highly competent strains of P. furiosus have been developed and thosehave been transformed at frequencies much higher than wild type (e.g.,DSM3638). Isolated P. furiosus of the invention are suitable to undergodiverse, industrially useful modifications, for the controlled production ofenzymes, production of chemicals (including H2).
Advantagesand Some Potential Applications
Facile and reproducible leading to a very high titer oftransformants
Applicable to other hyperthermophiles and, potentially,to other extremophiles
Provides strains (e.g. COM1) that are suitable to undergosubsequent modifications of interest to the end user
Inventors
Prof. Michael W.W. Adams and Prof JanetWestpheling
TechnologyDevelopment and IP Status
Lab scale (prototype stage)
Issued US Patent 8,927,254
- Country/Region
- USA
