The 3-Methyl-2-benzothiazolone Hydrazone Test.  Sensitive New Methods for the Detection, Rapid Estimation, and Determination of Aliphatic Aldehydes

Sawîckî, Eugene; Hauser, Thomas R.; Stanley, T. W.; Elbert, W.C.

doi:10.1021/ac60169a028

Cited by 402 publications

(61 citation statements)

References 12 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The assay was adapted from the procedure described in (25). Twenty Ixl of solutions of glutamate 1-semialdehyde were mixed with 100 pl of 0.4% 3-methyl-2-benzothiazolone hydrazone.…”

Section: Assay Of Aldehyde Functionmentioning

confidence: 99%

See 1 more Smart Citation

Biosynthesis of Δ-aminolevulinate in greening barley leaves. IX. Structure of the substrate, mode of gabaculine inhibition, and the catalytic mechanism of glutamate 1-semialdehyde aminotransferase

Hoober

Kahn

Ash

et al. 1988

Carlsberg Res. Commun.

101

View full text Add to dashboard Cite

Glutamic acid 1-semialdehyde hydrochloride was synthesized and purified. Its prior structural characterization was extended and confirmed by ~H NMR spectroscopy and chemical analyses. In aqueous solution at pH 1 to 2 glutamic acid 1-semialdehyde exists in a stable hydrated form, but at pH 8.0 it has a half-life of 3 to 4 rain. Spontaneous degradation of the material at pH 8.0 generated some undefined condensation products, but coincidentally a significant amount isomerized to 5-aminolevulinate. At pH 6.8 to 7.0, glutamate 1-semialdehyde is sufficiently stable to permit routine and reproducible assay for glutamate l-semialdehyde aminotransferase activity. Only about 20% of the enzyme extracted from chloroplasts was sensitive to inactivation by gabaculine with no pretreatment. However, when the enzyme was exposed to 5-aminolevulinate, levulinate or 4,5-dioxovalerate in the abs~ence of glutamate I-semialdehyde, it was completely inactivated by gabaculine; 4,6-dioxoheptanoate had no effect on the enzyme. These results lead to the hypothesis that the aminotransferase exists in the chloroplast in a complex with pyridoxamine phosphate, which must be converted to the pyridoxal form before it can form a stable adduct with gabaculine. We propose that the enzyme catalyzes the conversion of glutamate 1-semialdehyde to 5-aminolevulinate via 4,5-diaminovalerate.

show abstract

“…The assay was adapted from the procedure described in (25). Twenty Ixl of solutions of glutamate 1-semialdehyde were mixed with 100 pl of 0.4% 3-methyl-2-benzothiazolone hydrazone.…”

Section: Assay Of Aldehyde Functionmentioning

confidence: 99%

“…Elution ofglutamic acid 1-semialdehyde from Dowex 50W-X8 columns was detected by its reaction with the aldehyde-specific reagent 3-methyl-2-benzothiazolone hydrazone (25), reaction with ninhydrin, and enzymatic conversion to 5-aminolevulinate. analyzer.…”

Section: Properties Of Glutamic Acid L-semialdehydementioning

confidence: 99%

Biosynthesis of Δ-aminolevulinate in greening barley leaves. IX. Structure of the substrate, mode of gabaculine inhibition, and the catalytic mechanism of glutamate 1-semialdehyde aminotransferase

Hoober

Kahn

Ash

et al. 1988

Carlsberg Res. Commun.

101

View full text Add to dashboard Cite

show abstract

“…The control columns contained gel beads without immobilized material. The FA concentrations were tested for about three weeks in the outlet gas phase with a Formaldehyde Gas Detector (Model FP-40 Riken Keiki, Japan) and also in the aqueous column phase by a standard photometric method using a reaction with 1% chromotropic acid (Sawicki et al, 1961), as well as by the amperometric FdDHbased biosensor (Sigawi, 2010).…”

Section: Fa Removal From Indoor Airmentioning

confidence: 99%

Formaldehyde Oxidizing Enzymes and Genetically Modified Yeast Hansenula polymorpha Cells in Monitoring and Removal of Formaldehyde

Sibirny¹,

Demkiv²,

Sigawi³

et al. 2011

Waste Water - Evaluation and Management

View full text Add to dashboard Cite

“…Cell population densities were determined with a Coulter Counter (model ZBI, Coulter Electronics). The concentration of GSA was estimated by the modified 3-methyl-2-benzothiazolone assay (10,15).…”

Section: Other Proceduresmentioning

confidence: 99%

Biosynthesis of Tetrapyrrole Pigment Precursors

Avissar

Beale²

1989

Plant Physiol.

View full text Add to dashboard Cite

The aminotransferase that catalyzes the formation of 6-aminolevulinic acid from glutamate-1-semialdehyde or from glutamate in a reconstituted enzyme system was isolated and partially purified from Chlorella vulgaris. The apparent molecular weight of the aminotransferase was determined by Sephadex G-100 and Ultrogel AcA 54 gel filtration to be 60,000 ± 5,000. Catalytic activity of the aminotransferase required pyrixodal phosphate (PALP). The cofactor could not be removed by gel filtration after exposure of the enzyme to PALP. Aminotransferase was inhibited by gabaculine (3-amino-2,3-dihydrobenzoic acid). The concentration of gabaculine required for half maximal inhibition was about 0.05 micromolar. Aminotransferase activity could be regained upon the removal of gabaculine by gel filtration and supplementing the assay medium with PALP. Neither the inhibitory action of gabaculine nor its reversibility was affected by preincubation of the enzyme with the keto acids levulinate and &-aminolevulinic acid.the reaction to inhibitors that are considered to be pyridoxal antagonists, such as gabaculine and aminooxyacetate (5, 10, 13, 21). However, previous attempts to demonstrate directly a dependence of enzyme activity on PALP were unsuccessful (6,20). The apparent lack of dependence of the aminotransferase on PALP was explained as resulting from the existence of a tightly bound pyridoxal or pyridoxamine phosphate that is not removed from the enzyme by the various purification procedures employed during the preparation of the enzyme extract (5).We now report the direct demonstration of the dependence of aminotransferase activity in vitro on added PALP and the positive effect ofPALP on the recovery ofaminotransferase activity following preincubation of the enzyme with gabaculine.

show abstract

The 3-Methyl-2-benzothiazolone Hydrazone Test. Sensitive New Methods for the Detection, Rapid Estimation, and Determination of Aliphatic Aldehydes

Abstract: Sensitive new analytical methods for water-soluble aliphatic aldehydes has been introduced. Versatile spot plate, paper, silica gel, and colorimetric modifications of the procedure are of

Cited by 402 publications

References 12 publications

Biosynthesis of Δ-aminolevulinate in greening barley leaves. IX. Structure of the substrate, mode of gabaculine inhibition, and the catalytic mechanism of glutamate 1-semialdehyde aminotransferase

Biosynthesis of Δ-aminolevulinate in greening barley leaves. IX. Structure of the substrate, mode of gabaculine inhibition, and the catalytic mechanism of glutamate 1-semialdehyde aminotransferase

Formaldehyde Oxidizing Enzymes and Genetically Modified Yeast Hansenula polymorpha Cells in Monitoring and Removal of Formaldehyde

Biosynthesis of Tetrapyrrole Pigment Precursors

Contact Info

Product

Resources

About