Soluble reduced nicotinamide adenine dinucleotide oxidase from Bacillus cereus T spores and vegetative cells. II. Properties

1969 ◽  
Vol 15 (11) ◽  
pp. 1313-1317 ◽  
Author(s):  
D. H. Ashton ◽  
L. C. Blankenship
Keyword(s):  
Bacillus Cereus ◽  
Nicotinamide Adenine Dinucleotide ◽  
Vegetative Cell ◽  
Thermal Inactivation ◽  
Dipicolinic Acid ◽  
Heat Treatments ◽  
Nicotinamide Adenine ◽  
Flavin Mononucleotide ◽  
Vegetative Cells ◽  
Reduced Nicotinamide Adenine Dinucleotide

Two soluble reduced nicotinamide adenine dinucleotide (NADH2) oxidases purified from extracts of Bacillus cereus T spores were compared with vegetative ceil soluble NADH2 oxidase. The minor spore component and vegetative cell soluble NADH2 oxidase reacted equally well with riboflavin or flavin mononucleotide (FMN), were inhibited by 15 mM dipicolinic acid (DPA), and possessed similar thermal inactivation characteristics at 80 °C. Activity of the major spore component was stimulated by a factor of 3.6 when riboflavin replaced FMN as the coenzyme. The major spore component was not inhibited by DPA and resisted heat treatments which inactivated vegetative cell soluble NADH2 oxidase. These observations indicate that the minor spore component and vegetative cell soluble NADH2 oxidase are identical while the major spore component is a distinct protein.

Soluble reduced nicotinamide adenine dinucleotide oxidase from Bacillus cereus T spores and vegetative cells. I. Purification

1969 ◽  
Vol 15 (11) ◽  
pp. 1309-1312 ◽  
Author(s):  
D. H. Ashton ◽  
L. C. Blankenship
Keyword(s):  
Bacillus Cereus ◽  
Nicotinamide Adenine Dinucleotide ◽  
Gel Electrophoresis ◽  
Oxidase Activity ◽  
Vegetative Cell ◽  
Nicotinamide Adenine ◽  
Vegetative Cells ◽  
Cell Extracts ◽  
Cell Enzyme ◽  
Reduced Nicotinamide Adenine Dinucleotide

Spores and vegetative cells of Bacillus cereus T were disrupted by two procedures and soluble extracts prepared from the ruptured cells. Reduced nicotinamide adenine dinucleotide (NADH2) oxidases were purified from the extracts by ammonium sulfate fractionation, ion exchange on hydroxylapatite, and preparative acrylamide gel electrophoresis. The electrophoresis step revealed the presence of two distinct components with NADH2 oxidase activity in soluble extracts of spores while vegetative cell extracts contained only one. The faster moving component in spore extracts constituted about 30% of the NADH2 oxidase activity recovered and was identical with the vegetative cell enzyme in electrophoretic mobility on acrylamide gel. The slower moving spore component accounted for 70% of recovered activity and was found in soluble extracts regardless of the procedure used to rupture spores.

A disulfide reductase in spores of Bacillus cereus T

1971 ◽  
Vol 17 (10) ◽  
pp. 1273-1277 ◽  
Author(s):  
Leroy C. Blankenship ◽  
J. R. Mencher
Keyword(s):  
Molecular Weight ◽  
Bacillus Cereus ◽  
Nicotinamide Adenine Dinucleotide ◽  
Nicotinamide Adenine Dinucleotide Phosphate ◽  
Nicotinamide Adenine ◽  
Small Molecular Weight ◽  
Ph Optimum ◽  
Nitrobenzoic Acid ◽  
Disulfide Reductase ◽  
Reduced Nicotinamide Adenine Dinucleotide

An enzyme obtained from Bacillus cereus T spores which catalyzes the reduction of the disulfide, 5, 5′-dithiobis (2-nitrobenzoic acid) (DTNB), has been partially purified and characterized. The enzyme required either reduced nicotinamide adenine dinucleotide phosphate (NADPH2) or reduced nicotinamide adenine dinucleotide (NADH2) as electron donor. It had a pH optimum of 8, was destroyed by heating at 70C for 5 min, and was stimulated by Ca2+ and Mg2+. No other small molecular weight disulfides were found to be substrates for the enzyme.

Sign in / Sign up

Export Citation Format

Share Document