OxlT sequence and its use

Chemistry: molecular biology and microbiology – Micro-organism – per se ; compositions thereof; proces of... – Bacteria or actinomycetales; media therefor

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4353201, 435325, 435193, 435232, 536 231, 536 232, 536 237, C12N 120

Patent

active

061626325

ABSTRACT:
A DNA sequence encoding a transport protein responsible for the physical movement of oxalate and formate across the plasma membrane is provided. The protein, OxlT, can be isolated from the gram negative anaerobe, Oxalobacter formigenes. The protein is part of the three protein oxalate/formate antiport exchange system that allows for conversion of extracellular oxalate to formate+CO.sub.2.
The OxlT coding sequence can be used to provide a metabolic energy system to the cell. The system comprises the OxlT transport protein in combination with oxalylCoA decarboxylase and formyl-CoA transferase. Expression cassettes can be designed to express all or part of the system in a target host. Such energy generating systems can be used in industrial settings that utilize fermentation. The energy generating systems of the invention extend culture longevity and increase product yield.
In another aspect of the invention, the OxlT coding sequence can be used in pharmaceutical compositions for manipulating serum oxalic acid levels in patients in need thereof. In this manner, liposomes comprising the energy generating system can be developed and administered to humans and other animals.

REFERENCES:
H.L. Lung et al., "Cloning and Expression of the Oxalyl-CoA Decarboxylase Gene From the Bacterium, Oxalobacter formigenes: Prospects for Gene Therapy to Control Ca-Oxalate Kidney Stone Formation", Amer. J. Kidney Diseases XVII(4): 381-385, Apr. 1991.
K. Abe et al. "Exchange of Aspartate and Alanine", J.Biol. Chem. 271(6): 3079-3084, Feb. 1996.
Laine et al., "Stable Transformation of the Gram-Positive Phytopathogenic Bacterium Clavibacter michiganensis subsp. sepedonicus with Several Cloning Vectors," Applied and Environmental Microbiology, vol. 62, No. 5 (May 1996), pp. 1500-1506.
Fu et al. "Evaluation of Secondary Structure of OxlT, the Oxalate Transporter of Oxalobacter formigenes, by Circular Dichroism Spectroscopy," The Journal of Biological Chemistry, vol. 272, No. 4 (Jan. 24, 1997), pp. 2129-2135.
Ruan et al. "Identification, Purification, and Reconstitution of OxlT, the Oxalate:Formate Antiport Protein of Oxalobacter formigenes," The Journal of Biological Chemistry, vol. 267, No. 15 (May 25, 1992), pp. 10537-10543.

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