The isolation and characterization of 3-phosphoglycerate dehydrogenase from peas

1. 3-Phosphoglycerate dehydrogenase was purified 400-fold from crude extracts of etiolated pea epicotyls. 2. Michaelis constants were determined for all four substrates. 3. Loss of sensitivity to inhibition by l-serine occurs on purification. 4. The purified enzyme is inhibited by thiol-group reagents and, with N-ethyl-maleimide, protection is afforded by 3-phosphoglycerate though not by NAD+.

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Isolation and characterization of glyoxylate dehydrogenase from the fungus Sclerotium rolfsii

Biochemical Journal ◽

10.1042/bj2180113 ◽

1984 ◽

Vol 218 (1) ◽

pp. 113-118 ◽

Cited By ~ 23

Author(s):

A J Balmforth ◽

A Thomson

Keyword(s):

Affinity Chromatography ◽

Active Site ◽

Sclerotium Rolfsii ◽

Thiol Group ◽

Isolation And Characterization ◽

Crude Extracts ◽

Nad Oxidoreductase ◽

Inhibition Studies ◽

Dye Affinity Chromatography

Glyoxylate dehydrogenase (glyoxylate: NAD+ oxidoreductase) was purified 600-fold in three steps from crude extracts of the fungus Sclerotium rolfsii (Corticium rolfsii Curzi). Two of the purification steps involved dye-affinity chromatography. The enzyme is a tetramer of Mr 250 000, with identical subunits of Mr 57 000. Inhibition studies suggest that there is one essential thiol group per active site.

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Isolation and characterization of a low-molecular-weight substance activating head and bud formation in hydra

Development ◽

10.1242/dev.29.1.27 ◽

1973 ◽

Vol 29 (1) ◽

pp. 27-38

Author(s):

H. Chica Schaller

Keyword(s):

Molecular Weight ◽

Low Molecular Weight ◽

Bud Formation ◽

Isolation And Characterization ◽

Crude Extracts ◽

Head Formation ◽

Weight Substance

From crude extracts of hydra, a substance activating head formation was isolated and enriched at least 100000-fold. The molecular weight was determined to be approximately 900. Sensitivity against proteases suggests that it is a peptide. The substance acts at a concentration equivalent to the extract of 1 hydra per ml or at a concentration of less than 10 10M. In its highly purified form the substance activates head and bud formation.

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The purification and characterization of acetoacetyl-coenzyme A reductase from Azotobacter beijerinckii

Biochemical Journal ◽

10.1042/bj1210309 ◽

1971 ◽

Vol 121 (2) ◽

pp. 309-316 ◽

Cited By ~ 48

Author(s):

G. A. F. Ritchie ◽

P. J. Senior ◽

E. A. Dawes

Keyword(s):

Substrate Specificity ◽

Reaction Rate ◽

Acyl Carrier Protein ◽

Thiol Group ◽

Carrier Protein ◽

Purification And Characterization ◽

Michaelis Constants ◽

Coa Reductase ◽

Acetoacetyl Coa Reductase

A soluble acetoacetyl-CoA reductase (EC 1.1.1.36) was purified 54-fold from Azotobacter beijerinckii N.C.I.B. 9067 and the reaction product identified as d(−)-β-hydroxybutyryl-CoA. The Michaelis constants for acetoacetyl-CoA, NADPH and NADH were determined and the reaction rate was found to be some fivefold greater with NADPH than with NADH. At neutral pH the equilibrium greatly favours the formation of the reduced product. Substrate specificity was in the order: acetoacetyl-CoA>acetoacetylpantetheine>acetoacetyl-(acyl-carrier protein). The enzyme possesses a functional thiol group, suffers inactivation by oxygen and is inhibited by thiol-blocking reagents. Inhibition by p-chloromercuribenzoate is reversed by excess of dithiothreitol, which also protects the enzyme from inactivation by oxygen.

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Isolation and Characterization of Marine Endophytic Fungi from Seaweeds, and Bioactivity of their Crude Extracts

Journal of Pure and Applied Microbiology ◽

10.22207/jpam.13.3.15 ◽

2019 ◽

Vol 13 (3) ◽

pp. 1451-1460 ◽

Cited By ~ 1

Author(s):

Feroze Ahamed ◽

Marudhamuthu Murugan

Keyword(s):

Endophytic Fungi ◽

Isolation And Characterization ◽

Crude Extracts

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HPLC and NMR Studies of Phenoxazone Alkaloids from Pycnoporus Cinnabarinus

Natural Product Communications ◽

10.1177/1934578x0900400409 ◽

2009 ◽

Vol 4 (4) ◽

pp. 1934578X0900400 ◽

Cited By ~ 2

Author(s):

Daniel A. Dias ◽

Sylvia Urban

Keyword(s):

Antitumor Activity ◽

Spectroscopic Characterization ◽

2D Nmr ◽

Nmr Studies ◽

Pycnoporus Cinnabarinus ◽

Ergosterol Peroxide ◽

Isolation And Characterization ◽

Crude Extracts ◽

Antiviral Activities

Crude extracts of the red-orange, bracket fungus Pycnoporus cinnabarinus, collected from five distinct Australian localities were subjected to a chemical and biological profiling study. Subsequent detailed investigation of two of these specimens resulted in the isolation of the new phenoxazone alkaloid, pycnoporin (8), together with cinnabarin (1), tramesanguin (2), and cinnabarinic acid (3). Ergosterol peroxide (11) was also identified from one of the specimens studied. Compounds 1-3 and 8 were elucidated by detailed spectroscopic analyzes, which included the application of elevated temperature-controlled NMR experiments. In addition to the isolation and characterization of 8, this study describes the first successful HPLC purification strategy and complete 2D NMR spectroscopic characterization of compounds 1-3. Also reported are the antioxidant and anti-inflammatory activities of the crude extracts of one of the P. cinnabarinus specimens. Compounds 1-3, 8 and 11 displayed varying degrees of antitumor activity while ergosterol peroxide (11) also showed slight antimicrobial and antiviral activities. This is the first report documenting the significant antitumor activity of cinnabarin (1).

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