Studies on the biosynthesis of bialaphos (SF-1293). Part X. Purification and characterization of citrate synthase from Streptomyces hygroscopicus SF-1293 and comparison of its properties with those of 2-phosphinomethylmalic acid synthase.

Shimotohno, K; Imai, Shoichi; Murakami, Takeshi; Seto, Haruo

doi:10.1271/bbb1961.54.463

Cited by 10 publications

(6 citation statements)

References 2 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Furthermore, CitA subunits migrated with an apparent molecular mass on SDS-PAGE (45 kDa) (Fig. 1) consistent with that described for other citrate synthases (19,26,36,39). In order to isolate the citA gene, purified CitA was subjected to sequential Edman N-terminal degradation.…”

Section: Resultssupporting

confidence: 72%

“…Reaction kinetics of CitA-catalyzed conversion of acetyl-CoA and oxaloacetic acid to citrate were analyzed as described previously (13a). The apparent K m values for acetyl-CoA (32 M) and oxaloacetate (17 M) were similar to those citrate synthases from other gram-positive bacteria and eukaryotes but were quite distinct from the low affinities of citrate synthases from gram-negative organisms (i.e., GltA from E. coli and citrate synthase isoenzyme I from Pseudomonas aeruginosa (7,19,26,36,39).…”

Section: Resultsmentioning

confidence: 69%

“…S. coelicolor CitA activity was also unaffected by NAD ϩ (5 mM), ADP (5 mM), isocitrate (10 mM), or EDTA (0.05 mM). Interestingly, and in contrast to the citrate synthase of S. hygroscopicus (39), S. coelicolor CitA activity was not inhibited by ␣-ketoglutarate (10 mM).…”

Section: Resultsmentioning

confidence: 88%

“…Inhibition of S. coelicolor CitA by ATP was less pronounced (40% inhibition at 10 mM ATP) (19,20) than that reported for B. subtilis CitZ (50% inhibition at 0.81 mM ATP). Likewise, citrate synthases of S. hygroscopicus and C. glutamicum are only weakly inhibited by ATP (7,39). S. coelicolor CitA activity was also unaffected by NAD ϩ (5 mM), ADP (5 mM), isocitrate (10 mM), or EDTA (0.05 mM).…”

Section: Resultsmentioning

confidence: 99%

“…1). Ionexchange chromatography (DEAE, Q-Sepharose, MonoQ), followed by affinity chromatography (Matrex gel red A) (19,26,36,39), enriched the enzyme ca. 70-fold (Table 1).…”

Section: Resultsmentioning

confidence: 99%

See 4 more Smart Citations

Role of Acid Metabolism in Streptomyces coelicolor Morphological Differentiation and Antibiotic Biosynthesis

et al. 2001

View full text Add to dashboard Cite

Studies of citrate synthase (CitA) were carried out to investigate its role in morphological development and biosynthesis of antibiotics in Streptomyces coelicolor. Purification of CitA, the major vegetative enzyme activity, allowed characterization of its kinetic properties. The apparent K m values of CitA for acetyl coenzyme A (acetyl-CoA) (32 M) and oxaloacetate (17 M) were similar to those of citrate synthases from other gram-positive bacteria and eukaryotes. CitA was not strongly inhibited by various allosteric feedback inhibitors (NAD ؉ , NADH, ATP, ADP, isocitrate, or ␣-ketoglutarate). The corresponding gene (citA) was cloned and sequenced, allowing construction of a citA mutant (BZ2). BZ2 was a glutamate auxotroph, indicating that citA encoded the major citrate synthase allowing flow of acetyl-CoA into the tricarboxylic acid (TCA) cycle. Interruption of aerobic TCA cycle-based metabolism resulted in acidification of the medium and defects in morphological differentiation and antibiotic biosynthesis. These developmental defects of the citA mutant were in part due to a glucose-dependent medium acidification that was also exhibited by some other bald mutants. Unlike other acidogenic bald strains, citA and bldJ mutants were able to produce aerial mycelia and pigments when the medium was buffered sufficiently to maintain neutrality. Extracellular complementation studies suggested that citA defines a new stage of the Streptomyces developmental cascade.

show abstract

Section: Resultssupporting

confidence: 72%

Section: Resultsmentioning

confidence: 69%

Section: Resultsmentioning

confidence: 88%

Section: Resultsmentioning

confidence: 99%

“…1). Ionexchange chromatography (DEAE, Q-Sepharose, MonoQ), followed by affinity chromatography (Matrex gel red A) (19,26,36,39), enriched the enzyme ca. 70-fold (Table 1).…”

Section: Resultsmentioning

confidence: 99%

See 3 more Smart Citations

Role of Acid Metabolism in Streptomyces coelicolor Morphological Differentiation and Antibiotic Biosynthesis

et al. 2001

View full text Add to dashboard Cite

show abstract

Identification and characterization of a novel citrate synthase from Streptomyces diastaticus No. 7 strain M1033

Cao

Song

et al. 2015

Biotech and App Biochem

View full text Add to dashboard Cite

Citrate synthase (CS) is a key enzyme of the tricarboxylic acid cycle and is widely distributed among prokaryotes and eukaryotes. Here, we report for the first time, the cloning, expression, and characterization of a novel CS from Streptomyces diastaticus No. 7 strain M1033 (SdCS). Gel filtration chromatography and matrix-assisted laser-desorption ionization-time-of-flight mass spectrometry (MALDI-TOF-MS) analyses indicate that SdCS forms homodimers with a molecular mass of approximately 100.0 kDa. The predicted amino acid sequence of SdCS is highly similar to those of bacterial homodimeric type I CSs. The pH and temperature optima for SdCS activity were 8.0 and 35 °C, respectively. The half-life (t1/2 ) of SdCS was 10 Min at 50 °C and was increased to 210 Min in the presence of oxaloacetate. The kinetic parameters of SdCS (kcat = 262.8 and 230.7 s(-1); Km = 58.4 and 11.2 µM for acetyl-CoA and oxaloacetate, respectively) were comparable to those of dimeric CSs isolated from Gram-positive bacteria and eukaryotes. Moreover, SdCS activity was inhibited by ATP and ADP and stimulated by AMP. These findings provide a foundation for further investigations on the three-dimensional structure and mechanism of catalysis of SdCS.

show abstract

Ecophysiological studies on citrate-synthase: (I) enzyme regulation of selected crustaceans with regard to temperature adaptation

Vetter

1995

J Comp Physiol B

View full text Add to dashboard Cite

Studies on the biosynthesis of bialaphos (SF-1293). Part X. Purification and characterization of citrate synthase from Streptomyces hygroscopicus SF-1293 and comparison of its properties with those of 2-phosphinomethylmalic acid synthase.

Cited by 10 publications

References 2 publications

Role of Acid Metabolism in Streptomyces coelicolor Morphological Differentiation and Antibiotic Biosynthesis

Role of Acid Metabolism in Streptomyces coelicolor Morphological Differentiation and Antibiotic Biosynthesis

Identification and characterization of a novel citrate synthase from Streptomyces diastaticus No. 7 strain M1033

Ecophysiological studies on citrate-synthase: (I) enzyme regulation of selected crustaceans with regard to temperature adaptation

Contact Info

Product

Resources

About