Characterization of the glycolytic enzyme enolase which is abundant in the hyperthermophilic archaeon, Pyrococcus furiosus
- Argonne National Lab., IL (United States)
- Georgia Univ., Athens, GA (United States)
High enolase activity, as measured by the conversion of 2-phosphoglycerate to phosphoenolphyruvate, was found in the cytoplasm of Pyrococcus (an anaerobic, hyperthermophilic archaeon that grows optimally at 100{degree}C). In this organism, the enzyme probably functions in a sugar fermentation pathway. The enzyme was purified to homogeneity. It had a temperature optimum of >90 {degree}C, and a pH optimum of 8.1. The enzyme was extremely thermostable with a half time for inactivation at 100{degree}C of 40 min. In contrast, an enolase from yeast was inactivated in 1 min at 88{degree}C. Both the P. furiosus and yeast enzymes required a metal ion for activity, but whereas the yeast enzyme has an absolute requirement for Mg{sup ++} the P. furiosus enolase was equally active in the presence of Mn{sup ++}. Both enzymes were competitively inhibited by citrate. P. furiosus enolase, as for mesophilic enolases, probably has a homodimeric structure with subunit M{sub r} greater than 45,000. A highly conserved sequence of eight amino acids in the N-terminal region was found in enolases from P. furiosus and a wide range of other organisms including bacteria, yeast, birds, and mammals.
- Research Organization:
- Argonne National Lab. (ANL), Argonne, IL (United States)
- Sponsoring Organization:
- USDOE, Washington, DC (United States); Department of Defense, Washington, DC (United States)
- DOE Contract Number:
- W-31109-ENG-38
- OSTI ID:
- 10124321
- Report Number(s):
- ANL/CMB/PP-81754; ON: DE94006835; CNN: Grant N00014-90-J-1894
- Resource Relation:
- Other Information: PBD: [1993]
- Country of Publication:
- United States
- Language:
- English
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