|Formulation:||In 3.2 M ammonium sulphate|
|Stability:||Minimum 1 year at 4oC. Check vial for details.|
|Enzyme Activity:||Other Activities|
|Synonyms:||phosphate acetyltransferase; acetyl-CoA:phosphate acetyltransferase|
|Concentration:||Supplied at ~ 3,000 U/mL|
|Expression:||Recombinant from Bacillus Subtilis|
Catalyses the reaction:
Coenzyme A + acetyl phosphate = acetyl-CoA + phosphate
|Specific Activity:||~ 2,000 U/mg (25oC, pH 7.4 on Coenzyme A)|
|Unit Definition:||One Unit of phosphotransacetylase activity is defined as the amount of enzyme required to release one µmole of acetyl-CoA per minute from Coenzyme A (0.44 mM) in Tris.HCl buffer (86 mM) pH 7.4 at 25oC.|
|Application examples:||Applications for the measurement of acetic acid in the food, fermentation, wine, beverage and dairy industries.|
High purity recombinant phosphotransacetylase (Bacillus subtilis) for use in research, biochemical enzyme assays and in vitro diagnostic analysis.
See analytical enzymes for a list of all our available enzyme products.
Overall Retention of Methyl Stereochemistry during B12-Dependent Radical SAM Methyl Transfer in Fosfomycin Biosynthesis.
McLaughlin, M. I., Pallitsch, K., Wallner, G., van der Donk, W. A. & Hammerschmidt, F. (2021). Biochemistry, 60(20), 1587-1596.
Methylcobalamin-dependent radical S-adenosylmethionine (SAM) enzymes methylate non-nucleophilic atoms in a range of substrates. The mechanism of the methyl transfer from cobalt to the receiving atom is still mostly unresolved. Here we determine the stereochemical course of this process at the methyl group during the biosynthesis of the clinically used antibiotic fosfomycin. In vitro reaction of the methyltransferase Fom3 using SAM labeled with 1H, 2H, and 3H in a stereochemically defined manner, followed by chemoenzymatic conversion of the Fom3 product to acetate and subsequent stereochemical analysis, shows that the overall reaction occurs with retention of configuration. This outcome is consistent with a double-inversion process, first in the SN2 reaction of cob(I)alamin with SAM to form methylcobalamin and again in a radical transfer of the methyl group from methylcobalamin to the substrate. The methods developed during this study allow high-yield in situ generation of labeled SAM and recombinant expression and purification of the malate synthase needed for chiral methyl analysis. These methods facilitate the broader use of in vitro chiral methyl analysis techniques to investigate the mechanisms of other novel enzymes.Hide Abstract