What is the KM and Vmax of acid phosphatase?
The phosphatase had a Km of 0.24 mM and a Vmax of 2.76 μmol/min/mg with PLP.
What is the Vmax of alkaline phosphatase?
The Km and Vmax values were 0.5 x 10-3 M and 20 x 10-6 M/min, respectively. Inhibition studies showed that ALP activity was competitively inhibited by 0.67 mM sodium hydrogen orthophosphate (NaH2PO4) and the inhibition constant (Ki) was 0.9 x 10-3 M.
What is the KM value of alkaline phosphatase?
8.4 x 10
Alkaline phosphatase has a Km of 8.4 x 10−4.
What is the range of acid phosphatase enzyme in serum?
In adults and elderly persons, the normal findings for acid phosphatase are 0.13-0.63 U/L (Roy, Brower, Hayden; 37°C) or 2.2-10.5 U/L (SI units). Normal findings in children are 8.6-12.6 U/mL (30°C), while normal findings in newborns are 10.4-16.4 U/mL (30°C).
What does km measure in enzyme kinetics?
For practical purposes, Km is the concentration of substrate which permits the enzyme to achieve half Vmax. An enzyme with a high Km has a low affinity for its substrate, and requires a greater concentration of substrate to achieve Vmax.”
Why is Vmax rarely reached in physiological conditions?
Reaching Vmax requires saturating concentrations of substrate, and such high substrate concentrations are rarely reached in physiological conditions. Enzymes in physiological conditions are often binding alternative substrates, limiting the reaction velocity below Vmax for the preferred substrate.
How does alkaline phosphatase affect P nitrophenol?
p-Nitrophenol competitively inhibits the hydrolysis of p-nitrophenyl phosphate by alkaline phosphatase. Twenty microliters of enzyme preparation were added to 0.77, 0.193, 0.077, and 0.039 mM substrate in 7.0 ml of 1.0 M Tris-HCl buffer at pH 8.0 in the presence or absence of 30 μM p-nitrophenol.
What is P NPP?
PNPP (p-Nitrophenyl Phosphate, Disodium Salt) is a widely used substrate for detecting alkaline phosphatase in ELISA applications. When alkaline phosphatase and PNPP are reacted, a yellow water-soluble reaction product is formed. This reaction product absorbs light at 405nm. The substrate is offered in several formats.
What is normal range of serum alkaline phosphatase in adults?
The normal range for serum ALP level is 20 to 140 IU/L , but this can vary from laboratory to laboratory.
What is the normal value of prostatic ACP?
0 to 0.6 U/L
Prostatic AP = 0 to 0.6 U/L.
What does acid phosphatase measure?
Acid phosphatase is a ubiquitous lysosomal enzyme that hydrolyses organic phosphates at an acid pH. A variety of enzymatic and immunologic techniques have been employed to measure acid phosphatase. In the past, various substrates and inhibitors were utilized to increase specificity and sensitivity.
How is the km and Vmax of the acidic phosphatase determined?
Figure 4-1 Acidic phosphatase catalyzes the reaction Because of the hyperbolic shape of Vo against [S] plots, Vmax can only be determined from an extrapolation of the asymptotic approach of Vo to some limiting value as [S] increases indefinitely; and Km is derived from that value of [S] giving Vo = Vmax/2.
What is the maximum activity of a phosphatase?
PRINCIPLE: Phosphatases are enzymes that hydrolyze phosphate monoesters and release inorganic phosphate. Alkaline phosphatases (orthophosphoric monoester phosphohydrolases) are the 1 3. phosphate hydrolases that have a maximum activity at a relatively high pH (>7.0). This enzyme is widespread and occurs in both eukaryotic and prokaryotic cells.
Where can you find alkaline phosphatase in the body?
ALKALINE PHOSPHATASE ENZYME Alkaline phosphatase is a ubiquitous enzyme that can be isolated from bone, kidney, intestine, plasma, liver, spleen, plants and microorganisms. It catalyzes the removal of a phosphate that is attached to an alcohol, resulting in the generation of a free phosphate (Pi) and an alcohol.
How big is an alkaline phosphatase in daltons?
In this laboratory, we will be using alkaline phosphatase from bovine (cow) intestine. This particular alkaline phosphatase has a molecular weight of 138,000 daltons (a dalton is a unit of mass approximately equal to the mass of a hydrogen atom).