Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.1.4.3 (phospholipase C)
 18,461 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

A new colorimetric determination for serum phospholipid is described. Firstly, serum phospholipid is incubated with phospholipase C from Bacillus cereus, and then the released diglyceride and triglyceride are hydrolyzed completely to fatty acid and glycerol by lipoprotein lipase from Pseudomonas fluorescens. Secondly, the glycerol produced is enzymatically determined by glycerol dehydrogenase in the presence of NAD+, using phenazine methosulfate-nitro blue tetrazolium as color reagents. The absorbance at 570 nm is recorded. The amount of the glycerol from phospholipid is calculated by subtracting the amount of glycerol from triglyceride from the amount of total glycerol. The present method requires only 20 microliter of serum and a 40 min incubation and is highly reproducible. The results obtained show good correlation with those obtained by a chemical method (correlation coefficient, 0.925) or the phospholipase D-choline oxidase method (correlation coefficient, 0.936). These results strongly suggest that the proposed method can be utilized as a routine clinical test.
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PMID:An enzymic determination for serum phospholipid. 70 86

Two alternative procedures are described for the quantitative determination of phosphatidylcholine in a flow-injection system utilizing immobilized enzymes. Phospholipase C from Bacillus cereus and phospholipase D from cabbage were covalently bound to the surface of controlled-pore glass beads and the enzyme-derivatized beads were packed in small columns. In the first procedure, the phospholipase C column was connected with a second column containing coimmobilized alkaline phosphatase and choline oxidase. In the alternative procedure, the column packed with immobilized phospholipase D was connected with a column packed with immobilized choline oxidase. The hydrogen peroxide produced through the action of choline oxidase in both flow-injection systems was detected amperometrically. Both procedures are suitable for an accurate and rapid quantitation of phosphatidylcholine. The sensitivity of the method based on phospholipase C and alkaline phosphatase is higher than that using phospholipase D. Quantitation of phosphatidylcholine at the nanomole level can be easily obtained using the first method.
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PMID:Determination of phosphatidylcholine in a flow injection system using immobilized enzyme reactors. 220 Mar 5

We describe methods for automated enzymatic measurement of lecithin, sphingomyelin, and phosphatidylglycerol in amniotic fluid. Phospholipase C (EC 3.1.4.3) and sphingomyelin phosphodiesterase (EC 3.1.4.12) are reacted with lecithin and sphingomyelin, respectively, to liberate phosphocholine. Phosphocholine is then reacted with alkaline phosphatase, choline oxidase, peroxidase, and 4-aminoantipyrine to form a colored complex, for which the absorbance at 500 nm is measured with a centrifugal analyzer. Phosphatidylglycerol is hydrolyzed by phospholipase D (EC 3.1.4.4) to form glycerol, which is subsequently reacted with ATP and NAD+ in the presence of glycerol kinase and glycerol-3-phosphate dehydrogenase to yield NADH. The absorbance of the NADH formed is measured at 340 nm. These methods provide a simple, rapid, and accurate alternative to thin-layer chromatography for determination of phospholipids in amniotic fluid for assessment of fetal lung maturity.
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PMID:Automated enzymatic measurement of lecithin, sphingomyelin, and phosphatidylglycerol in amniotic fluid. 380 1

We describe a method for the direct enzymatic determination of phosphatidylcholine and sphingomyelin in serum. Phospholipase C (EC 3.1.4.3) and sphingomyelinase (EC 3.1.4.12) are used to hydrolyze phosphatidylcholine and sphingomyelin, respectively, with high specificity; alkaline phosphatase (EC 3.1.3.1) is used to cleave inorganic phosphorus. The choline group, after oxidation with choline oxidase (EC 1.1.3.17), is detected with 4-aminoantipyrine. Alternatively, phosphorus is assayed with metavanadate. The excellent agreement between results of this modification of a procedure described by Artiss et al. (Microchem. J. 26: 1017, 1980) for amniotic fluid and of the conventional thin-layer chromatographic method makes this an attractive method for determination of both phospholipid subclasses in serum.
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PMID:Enzymic assay for phosphatidylcholine and sphingomyelin in serum. 634 Aug 53

An economical enzymatic assay for HDL phosphatidyl choline is described, as adapted for the Cobas-Bio analyser (Hoffmann LaRoche). This method entails the enzymatic cleavage of phosphatidyl choline by phospholipase C from B. cereus, hydrolysis of phosphoryl choline and enzymatic determination of choline with choline oxidase by an enzymatic colour test. This method provides consistent values and is, by comparison to the enzymatic UV method (assaying choline with choline kinase in an optical test procedure), simpler to perform, more precise, and less expensive.
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PMID:An economical assay for HDL phosphatidyl choline. 635 3

A sensitive and selective flow injection method for the determination of phosphatidylcholine (PC) in sediment pore waters and extracts is described. It involves the use of phospholipase C, alkaline phosphatase and choline oxidase co-immobilized on controlled pore glass in a packed column reactor. The final product of the enzymatic reaction of phosphatidylcholine is hydrogen peroxide, and this is detected by measuring the chemiluminescence emission resulting from cobalt(II) catalysed reaction with luminol. The flow injection method is rapid (30 injections/h), reproducible (1.4% R.S.D. at 3 microM PC, n=10) with a detection limit of 0.14 microM (estimated from 3sigma(n-1) of the measured blank). A linear calibration response was obtained over a concentration range of 0.5-9 microM (r=0.999). The method has been applied to the determination of phosphatidylcholine in sediment extracts and sediment pore waters.
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PMID:An enzymatic flow analysis method for the determination of phosphatidylcholine in sediment pore waters and extracts. 1897 5

Pseudomonas aeruginosa can acquire and metabolize a variety of molecules including choline, an abundant host-derived molecule. In P. aeruginosa, choline is oxidized to glycine betaine which can be used as an osmoprotectant, a sole source of carbon and nitrogen, and as an inducer of the virulence factor, hemolytic phospholipase C (PlcH) via the transcriptional regulator GbdR. The primary objective was to determine the contribution of choline conversion to glycine betaine to P. aeruginosa survival during mouse lung infection. A secondary objective was to gain insight into the relative contributions of the different roles of glycine betaine to P. aeruginosa survival during infection. Using a model of acute murine pneumonia, we determined that deletion of the choline oxidase system (encoded by betBA) decreased P. aeruginosa survival in the mouse lung. Deletion of the glycine betaine demethylase genes (gbcA-B), required for glycine betaine catabolism, did not impact P. aeruginosa survival in the lung. Thus, the defect of the betBA mutant was not due to a requirement for glycine betaine catabolism or dependence on a downstream metabolite. Deletion of betBA decreased the abundance of plcH transcript during infection, which suggested a role for PlcH in the betBA survival defect. To test the contribution of plcH to the betBA mutant phenotype a betBAplcHR double deletion mutant was generated. The betBA and betBAplcHR double mutant had a small but significant survival defect compared to the plcHR single mutant, suggesting that regulation of plcH expression is not the only role for glycine betaine during infection. The conclusion was that choline acquisition and its oxidation to glycine betaine contribute to P. aeruginosa survival in the mouse lung. While defective plcH induction can explain a portion of the betBA mutant phenotype, the exact mechanisms driving the betBA mutant survival defect remain unknown.
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PMID:Choline catabolism to glycine betaine contributes to Pseudomonas aeruginosa survival during murine lung infection. 2345 28