Determination of the scissile bond in the hydrolysis of α-d-ribofuranose 1-phosphate by alkaline phosphatase, acid phosphatase and formic acid and in its conversion to d-ribose 5-phosphate by phosphoglucomutase

1982 ◽  
Vol 704 (3) ◽  
pp. 427-436 ◽  
Author(s):  
Frank Jordan ◽  
Donald J. Kuo ◽  
Salvatore J. Salamone ◽  
Alice L. Wang
Keyword(s):  
Alkaline Phosphatase ◽  
Acid Phosphatase ◽  
Formic Acid ◽  
Scissile Bond ◽  
Hydrolysis Of

scholarly journals Phosphatase synthesis in Klebsiella (Aerobacter) aerogenes growing in continuous culture

1972 ◽  
Vol 127 (1) ◽  
pp. 87-96 ◽  
Author(s):  
P. G. Bolton ◽  
A. C. R. Dean
Keyword(s):  
Alkaline Phosphatase ◽  
Acid Phosphatase ◽  
Continuous Culture ◽  
Activity Profile ◽  
Glucose Effect ◽  
Ph Values ◽  
Nitrophenyl Phosphate ◽  
Wide Range ◽  
Rate Of Hydrolysis ◽  
Hydrolysis Of

1. Phosphatase synthesis was studied in Klebsiella aerogenes grown in a wide range of continuous-culture systems. 2. Maximum acid phosphatase synthesis was associated with nutrient-limited, particularly carbohydrate-limited, growth at a relatively low rate, glucose-limited cells exhibiting the highest activity. Compared with glucose as the carbon-limiting growth material, other sugars not only altered the activity but also changed the pH–activity profile of the enzyme(s). 3. The affinity of the acid phosphatase in glucose-limited cells towards p-nitrophenyl phosphate (Km 0.25–0.43mm) was similar to that of staphylococcal acid phosphatase but was ten times greater than that of the Escherichia coli enzyme. 4. PO43−-limitation derepressed alkaline phosphatase synthesis but the amounts of activity were largely independent of the carbon source used for growth. 5. The enzymes were further differentiated by the effect of adding inhibitors (F−, PO43−) and sugars to the reaction mixture during the assays. In particular, it was shown that adding glucose, but not other sugars, stimulated the rate of hydrolysis of p-nitrophenyl phosphate by the acid phosphatase in carbohydrate-limited cells at low pH values (<4.6) but inhibited it at high pH values (>4.6). Alkaline phosphatase activity was unaffected. 6. The function of phosphatases in general is discussed and possible mechanisms for the glucose effect are outlined.

An Automated Alkaline Phosphatase Assay with Phenolphthalein Monophosphate as Substrate

1967 ◽  
Vol 13 (4) ◽  
pp. 281-289 ◽  
Author(s):  
Kirsten Hviid
Keyword(s):  
Alkaline Phosphatase ◽  
Normal Values ◽  
Serum Alkaline Phosphatase ◽  
Colorimetric Determination ◽  
Alkaline Phosphatase Assay ◽  
Phosphatase Assay ◽  
Hydrolysis Of

Abstract The manual procedure of Babson et al. (1) for the determination of serum alkaline phosphatase has been automated. The assay is based on the colorimetric determination of phenolphthalein formed on hydrolysis of phenolphthalein monophosphate. The procedure utilizes 0.16 ml. of serum without dialysis. Blanks are required only for turbid sera. Results are compared with those obtained by the manual procedure, and data relating to sample interaction, precision, blank values, and normal values are presented.

428. The acid phosphatase of mammary tissue of the cow and the rat

1950 ◽  
Vol 17 (3) ◽  
pp. 306-311 ◽  
Author(s):  
J. E. C. Mullen
Keyword(s):  
Alkaline Phosphatase ◽  
Acid Phosphatase ◽  
Replacement Therapy ◽  
Marked Effect ◽  
Mammary Tissue ◽  
Optimum Ph ◽  
Rate Of Hydrolysis ◽  
Hydrolysis Of

1. Mammary tissue of the cow and the rat contains acid phosphatase. The respective pH optima are 5·5–5·8 and 6·0.2. The enzyme in cow mammary tissue is probably one of type AII in the Folley & Kay (10) classification.3. The acid phosphatase of cow mammary tissue is inhibited by a factor in raw cows' milk. This factor is destroyed by heat.4. On the basis of rate of hydrolysis of phenylphosphate at the optimum pH there is about 5 times more alkaline phosphatase in the mammary tissue of the cow than acid phosphatase.5. The effect of adrenalectomy and replacement therapy through the administration of cortical steroids has no marked effect on the acid phosphatase of rat mammary tissue.

THE DETERMINATION OF TRYPTOPHANE BY A MODIFIED GLYOXYLIC ACID METHOD EMPLOYING PHOTOELECTRIC COLORIMETRY

1938 ◽  
Vol 16b (10) ◽  
pp. 361-368 ◽  
Author(s):  
J. L. D. Shaw ◽  
W. D. McFarlane
Keyword(s):  
Formic Acid ◽  
Sodium Hydroxide ◽  
Reliable Method ◽  
Glyoxylic Acid ◽  
Acid Method ◽  
Photoelectric Colorimeter ◽  
Acid Reaction ◽  
Alkali Hydrolysis ◽  
Hydrolysis Of

An accurate and reliable method for the estimation of tryptophane is described. It is based on the glyoxylic acid reaction, and involves the use of the Evelyn photoelectric colorimeter. The technique makes it possible to ascertain readily whether the color being measured is due only to tryptophane.The method has been applied to casein, of which, if necessary, only 25 mg. is required. The tryptophane determination is readily accomplished on a solution obtained by dissolving the casein in 10 or 20% sodium hydroxide or 5% formic acid by heating for a few minutes. With respect to alkali hydrolysis of casein under pressure, tryptophane is unstable in the sodium hydroxide hydrolysis, but is very stable in the baryta hydrolysis. The age and source of the casein are shown to be factors causing variations in the tryptophane content of different samples of casein, a variability which has been observed by a few previous workers.

A label-free conjugated polymer-based fluorescence assay for the determination of adenosine triphosphate and alkaline phosphatase

2014 ◽  
Vol 38 (9) ◽  
pp. 4574-4579 ◽  
Author(s):  
Yanan Li ◽  
Yan Li ◽  
Xinyan Wang ◽  
Xingguang Su
Keyword(s):  
Alkaline Phosphatase ◽  
Adenosine Triphosphate ◽  
Conjugated Polymer ◽  
Fluorescence Assay ◽  
Label Free ◽  
Quenching Effect ◽  
Hydrolysis Of

A sensor was developed based on the quenching effect of Cu2+ on PPESO3 and the hydrolysis of ATP by ALP.

scholarly journals Restoring the Oxidase-Like Activity of His@AuNCs for the Determination of Alkaline Phosphatase

Biosensors ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 174
Author(s):  
Fanfan Xiao ◽  
Yuting Yu ◽  
Yang Wu ◽  
Lili Tian ◽  
Guoyan Zhao ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Self Assembly ◽  
Colorimetric Method ◽  
Serum Samples ◽  
Turn On ◽  
Alp Activity ◽  
Assembly Method ◽  
Diallyldimethylammonium Chloride ◽  
Hydrolysis Of

In this paper, we propose a simple colorimetric method for the sensitive and selective detection of alkaline phosphatase (ALP) activity based on the turn off/turn on oxidase mimic activity of His@AuNCs. His@AuNCs/graphene oxide hybrids (His@AuNCs/GO) were easily obtained using the self-assembly method with poly (diallyldimethylammonium chloride) (PDDA)-coated GO and showed high oxidase-like activity compared with His@AuNCs. We found that the pyrophosphate ion (P2O74−, PPi) could effectively inhibit the oxidase mimic activity of His@AuNCs/GO, and the hydrolysis of PPi by ALP restored the inhibited activity of His@AuNCs/GO, enabling them to efficiently catalyze the oxidation of 3,3′,5,5′-tetramethylbenzidine (TMB) to generate the blue oxidized product oxTMB. The intensity of the color showed a linear dependency with the ALP activity. ALP was detected in the linear range of 0–40 mU/mL with a low detection limit (LOD) of 0.26 mU/mL (S/N = 3). The proposed method is fast, easy, and can be applied to monitor the ALP activity in serum samples accurately and effectively, which suggests its practicability and reliability in the detection of ALP activity in clinical practice.

New Substrate for Alkaline Phosphatase in Milk

1967 ◽  
Vol 50 (3) ◽  
pp. 555-557
Author(s):  
Arthur L Babson ◽  
Sharon J Greeley
Keyword(s):  
Alkaline Phosphatase ◽  
Organic Solvents ◽  
False Positive ◽  
Pasteurized Milk ◽  
False Positive Reactions ◽  
Hydrolysis Of

Abstract Phcnolphthalein monophosphale is used as a new substrate for the determination of residual alkaline phosphatase in pasteurized milk. Hydrolysis of this compound yields phcnolphthalein and thus a red solution which can be compared visually with a standard prepared from the same milk. The three reagents required are stable and can be delivered directly from plastic dropping bottles. No extraction into organic solvents is necessary, and this method eliminates the chances for false positive reactions

DETERMINATION OF MICRO-AMOUNTS OF 5β-PREGNAN-3α-OL-20-ONE IN URINE USING INFRARED-SPECTROMETRY

1962 ◽  
Vol 41 (2) ◽  
pp. 234-246 ◽  
Author(s):  
H. J. van der Molen
Keyword(s):  
Infrared Spectrum ◽  
Quantitative Determination ◽  
Acid Hydrolysis ◽  
Chromatographic Separation ◽  
Pure Form ◽  
Infrared Spectrometry ◽  
Hydrolysis Of

ABSTRACT A procedure for the quantitative determination of 5β-pregnan-3α-ol-20-one in urine is described. After acid hydrolysis of the pregnanolone-conjugates in urine, the free steroids are extracted with toluene. Pregnanolone is isolated in a pure form as its acetate; after chromatographic separation of the free steroids on alumina, the fraction containing pregnanolone is acetylated and rechromatographed on alumina. Quantitative determination of the isolated pregnanolone-acetate is carried out with the aid of the infrared spectrum recorded by a micro KBr-wafermethod. The reliability of the method under various conditions is discussed under the headings, specificity, accuracy, precision and sensitivity. It is possible to determine 30–40 μg pregnanolone in a 24-hours urine portion with a precision of 25%.

EINE CHEMISCHE METHODE ZUR BESTIMMUNG VON 16-EPI-ÖSTRIOL IM URIN DES MENSCHEN

1963 ◽  
Vol 44 (1) ◽  
pp. 47-66 ◽  
Author(s):  
W. Nocke ◽  
H. Breuer
Keyword(s):  
Melting Point ◽  
Phosphoric Acid ◽  
Aqueous Alkali ◽  
Percentage Error ◽  
Organic Layer ◽  
Maximum Percentage ◽  
Chemical Determination ◽  
Routine Assay ◽  
Hydrolysis Of

ABSTRACT A method for the chemical determination of 16-epi-oestriol in the urine of nonpregnant women with a qualitative sensitivity of less than 0.5 μg/24 h is described. The separation of 16-epi-oestriol and oestriol is accomplished by converting 16-epi-oestriol into its acetonide, a reaction which is stereoselective for cis-glycols and therefore not undergone by oestriol as a trans-glycol. Following partition between chloroform and aqueous alkali, the acetonide of 16-epi-oestriol is completely separated with the organic layer whereas oestriol as a strong phenol remains in the alkaline phase. 16-epi-oestriol is chromatographed on alumina as the acetonide and determined as a Kober chromogen. This procedure can easily be incorporated into the method of Brown et al. (1957 b) thus making possible the simultaneous routine assay of oestradiol-17β, oestrone, oestriol and 16-epi-oestriol from one sample of urine. The specificity of the method was established by separation of 16-epi-oestriol from nonpregnancy urine as the acetonide, hydrolysis of the acetonide by phosphoric acid, isolation of the free compound by microsublimation and identification by micro melting point, colour reactions and chromatography. The accuracy of the method is given by a mean recovery of 64% for pure crystalline 16-epi-oestriol when added to hydrolysed urine in 5–10 μg amounts. The precision is given by s = 0.24 μg/24 h. For the duplicate determination of 16-epi-oestriol the qualitative sensitivity is 0.44 μg/24 h, the maximum percentage error being ± 100% The quantitative sensitivity (±25% error) is 1.7 μg/24 h.

INDIREKTE PAPIERCHROMATOGRAPHISCHE METHODE ZUR BESTIMMUNG DER HARNOESTROGENE

1961 ◽  
Vol 38 (4) ◽  
pp. 545-562 ◽  
Author(s):  
L. Kecskés ◽  
F. Mutschler ◽  
I. Glós ◽  
E. Thán ◽  
I. Farkas ◽  
...  
Keyword(s):  
Pregnant Women ◽  
Granulosa Cell ◽  
Iron Content ◽  
List Type ◽  
Granulosa Cell Tumour ◽  
Hydrolysis Of ◽  
Phenol Fraction ◽  
Qualitative Determination

ABSTRACT 1. An indirect paperchromatographic method is described for separating urinary oestrogens; this consists of the following steps: acidic hydrolysis, extraction with ether, dissociation of phenol-fractions with partition between the solvents. Previous purification of phenol fraction with the aid of paperchromatography. The elution of oestrogen containing fractions is followed by acetylation. Oestrogen acetate is isolated by re-chromatography. The chromatogram was developed after hydrolysis of the oestrogens 'in situ' on the paper. The quantity of oestrogens was determined indirectly, by means of an iron-reaction, after the elution of the iron content of the oestrogen spot, which was developed by the Jellinek-reaction. 2. The method described above is satisfactory for determining urinary oestrogen, 17β-oestradiol and oestriol, but could include 16-epioestriol and other oestrogenic metabolites. 3. The sensitivity of the method is 1.3–1.6 μg/24 hours. 4. The quantitative and qualitative determination of urinary oestrogens with the above mentioned method was performed in 50 pregnant and 9 non pregnant women, and also in 2 patients with granulosa cell tumour.

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