Production of poly-beta-hydroxybutyrate in transformed escherichia coli
AssigneeCenter for Innovative Technology
InventorDouglas E. Dennis
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Abstract
Methods are provided for enhancing the production of PHB from a transformed E. coli host which includes the genes coding for the PHB biosynthetic pathway. By inserting the genes coding for PHB into a host which includes a lactose utilization system, a low cost minimal medium including whey can be used as the fuel and carbon source for PHB production. A plasmid which codes for the PHB biosynthetic pathway plus four hundred extra bases on either side of the first and last genes in the pathway has been inserted into the host and has been shown to produce a larger amount of PHB accumulation in a shorter period of time than other plasmid constructs. CaCl.sub.2 has been shown to be an effective agglomerating agent for agglomerating PHB which has been produced in a transformed E. coli host.
Claims15
An Escherichia coli bacterial host transformed by a vector consisting essentially of a deoxyribonucleic acid sequence encoding the poly-beta-hydroxybutyrate biosynthetic pathway of Alcaligenes eutrophus wherein the bacterial host has a lactose utilization system and grows in minimal medium containing whey and wherein the bacterial host is Escherichia coli strain ATCC 68329.
A method for agglomerating poly-beta-hydroxybutyrate which has been intracellularly produced in a culture of Escherichia coli (strain ATCC 68329) bacterial host which has been transformed by a vector the vector containing a deoxyribonucleic acid sequence coding for the poly-beta-hydroxybutyrate biosynthetic pathway from Alcaligenes eutrophus, comprising the steps of:
- lysing said bacterial hosts to release said poly-beta-hydroxybutyrate;
- and
- adding to said released poly-beta-hydroxybutyrate a sufficient quantity of an ionic reagent selected from the group consisting of magnesium sulfate, magnesium chloride, magnesium acetate and calcium chloride, said sufficient quantity of said ionic reagent agglomerating granules of said poly-beta-hydroxybutyrate.
A plasmid designated as p4A and deposited with the American Type Culture Collection in Escherichia coli strain HMS 174 under accession number 68329.
A plasmid designated as pTZ-18U PHB and deposited with the American Type Culture Collection in Escherichia coli strain HMS174 under accession number 69064.
Dependencies are inferred from the claim text.