Enzymatic degradation of cyclic 2,3-diphosphoglycerate to 2,3-diphosphoglycerate in Methanobacterium thermoautotrophicum
- Boston Coll., Chestnut Hill, MA (United States)
2,3-Diphosphoglycerate (2,3-DPG) has been found to be the product of the enzymatic degradation of cyclic 2,3-diphosphoglycerate (cDPG) in the archaebacterium Methanobacterium thermoautotrophicum {Delta}H. Although 2,3-DPG has not previously been detected as a major soluble component of M. thermoautotrophicum, large pools accumulated at an incubation temperature of 50C. Under these conditions, cellular activity was significantly decreased; a return of the culture to the optimum growth temperature restored the 2,3-DPG pool back to original low levels and caused steady-state cDPG levels to increase again. While {sup 13}CO{sub 2}-pulse/{sup 12}CO{sub 2}-chase experiments at 50C showed that the cDPG turned over, the appearance of 2,3-DPG at NMR-visible concentrations required at least 10 h. Production of 2,3-DPG in vivo was prevented by exposure of the cells to O{sub 2}. the enzyme responsible for this hydrolysis of cDPG was purified by affinity chromatography and appears to be a 33-kDa protein. Activity was detected in the presence of oxygen and was enhanced by a solution of 1 M KCl, 25 mM MgCl{sub 2}, and dithiothreitol. Both K{sub m} and V{sub max} have been determined at 37C; kinetics also indicate that in vitro the product, 2,3-DPG, is an inhibitor of cDPG hydrolysis. These findings are discussed in view of a proposed role for cDPG in methanogens.
- DOE Contract Number:
- FG02-91ER20025
- OSTI ID:
- 5640111
- Journal Information:
- Biochemistry; (United States), Journal Name: Biochemistry; (United States) Vol. 31:11; ISSN 0006-2960; ISSN BICHA
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
62 RADIOLOGY AND NUCLEAR MEDICINE
BACTERIA
BIOCHEMICAL REACTION KINETICS
BIODEGRADATION
CARBOHYDRATES
CARBON 12
CARBON 13
CARBON COMPOUNDS
CARBON DIOXIDE
CARBON ISOTOPES
CARBON OXIDES
CHALCOGENIDES
CHEMICAL REACTIONS
CHEMICAL SHIFT
DECOMPOSITION
ENZYMATIC HYDROLYSIS
ESTERS
EVEN-EVEN NUCLEI
EVEN-ODD NUCLEI
HYDROLYSIS
ISOTOPES
KINETICS
LIGHT NUCLEI
LIPIDS
LYSIS
MAGNETIC RESONANCE
METHANOGENIC BACTERIA
MICROORGANISMS
NUCLEAR MAGNETIC RESONANCE
NUCLEI
ORGANIC COMPOUNDS
ORGANIC PHOSPHORUS COMPOUNDS
OXIDES
OXYGEN COMPOUNDS
PHOSPHOLIPIDS
REACTION KINETICS
RESONANCE
SOLVOLYSIS
STABLE ISOTOPES