Esterase, pancreatic

PPL = porcine pancreatic lipase PLE = porcine liver esterase. 

Lippi et. al (87) and Dirstine (88) circumvented titration by converting the liberated fatty acids into copper salts, which after extraction in chloroform are reacted with diethyldithio-carbamate to form a colored complex which is measured photometrically. While the end point appears to be more sensitive than the pH end point determination, the advantages are outweighed by the additional steps of solvent extraction, centrifugation and incomplete extraction when low concentrations of copper salts are present. Other substrates used for the measurement of lipase activity have been tributyrin ( ), phenyl laurate (90), p-nit ro-pheny1-stearate and 3-naphthyl laurate (91). It has been shown that these substrates are hydrolyzed by esterases and thus lack specificity for lipase. Studies on patients with pancreatitis indicate olive oil emulsion is definitely superior to water soluble esters as substrates for measuring serum lipase activity. 

Song, H. Tietz, N. W. and Tan, C. Usefulness of serum lipase, esterase and amylase estimation in the diagnosis of pancreatitis - a comparison. Clin. 

A variety of enzymes (such as acetylcholine esterase, Porcine pancreatic lipase, Pseudomonas cepacia lipase, and Candida antarcita lipase) have been found useful in the preparation of enantiomerically pure cyclopentenol (+)-2 from 1. The enantiomeric (—)-2 has been prepared from diol 4 by enzymatic acetylation catalyzed by VP-345 with isopropenyl acetate in an organic medium. The key intermediate cyclopentanones (+)-6, (—)-6, 7, and 8, which are useful in the preparation of many bioactive molecules, can be obtained from 3 and 5 via routine chemical transformations.7... 

The overall metabolism of vitamin A in the body is regulated by esterases. Dietary retinyl esters are hydrolyzed enzymatically in the intestinal lumen, and free retinol enters the enterocyte, where it is re-esterified. The resulting esters are then packed into chylomicrons delivered via the lymphatic system to the liver, where they are again hydrolyzed and re-esterified for storage. Prior to mobilization from the liver, the retinyl esters are hydrolyzed, and free retinol is complexed with the retinol-binding protein for secretion from the liver [101]. Different esterases are involved in this sequence. Hydrolysis of dietary retinyl esters in the lumen is catalyzed by pancreatic sterol esterase (steryl-ester acylhydrolase, cholesterol esterase, EC 3.1.1.13) [102], A bile salt independent retinyl-palmitate esterase (EC 3.1.1.21) located in the liver cell plasma hydrolyzes retinyl esters delivered to the liver by chylomicrons. Another neutral retinyl ester hydrolase has been found in the nuclear and cytosolic fractions of liver homogenates. This enzyme is stimulated by bile salts and has properties nearly identical to those observed for... 

J. Hyun, C. R. Treadwell, G. V. Vahouny, Pancreatic Juice Cholesterol Esterase. Studies on Molecular Weigth and Bile Salt-Induced Polymerization , Arch. Biochem. Bio-phys. 1972, 752, 233-242. 

R. M. Stroud, Structure of Bovine Pancreatic Cholesterol Esterase at 1.6 A Novel Structural Features Involved in Lipase Activation , Biochemistry 1998, 37, 5107-5117. 

E. M. Kyger, D. J. S. Riley, C. A. Spilburg, L. G. Lange, Pancreatic Cholesterol Esterases. 3. Kinetic Characterization of Cholesterol Ester Resynthesis by Pancreatic Cholesterol Esterases , Biochemistry 1990, 29, 3853-3858. 

G. K. E. Scriba, Phenytoin-Lipid Conjugates Chemical, Plasma Esterase-Mediated, and Pancreatic Lipase-Mediated Hydrolysis in vitro, Pharm. Res. 1993, 10, 1181 — 1186. 

Various pancreatic enzymes hydrolyze lipids, including lipase with its auxiliary protein colipase (see p. 270), phospholipase A2, and sterol esterase. Bile salts activate the lipidcleaving enzymes through micelle formation (see below). 

Figure 11, Double kinetic resolution using enzymes with opposite enantioselectivity. PPL, porcine pancreatic lipase PLE, pig liver esterase.

Kinetic optical resolution of racemic alcohols and carboxylic acids by enzymatic acyl transfer reactions has received enormous attention in recent years56. The enzymes generally employed are commercially available lipases and esterases, preferentially porcine liver esterase (PLE) or porcine pancreatic lipase (PPL). Lipases from microorganisms, such as Candida cylindracea, Rhizopus arrhizus or Chromobacterium viscosum, are also fairly common. A list of suitable enzymes is found in reference 57. Standard procedures are described in reference 58. Some examples of the resolution of racemic alcohols are given39. 

Cholesteryl ester degradation Most dietary cholesterol is present in the free (nonesterified) form, with ten to fifteen percent present in the esterified form. Cholesteryl esters are hydrolyzed by pancreatic cholesterol ester hydrolase (cholesterol esterase), which produces cholesterol plus free fatty acids (see Figure 15.2). Cholesteryl esteh hydrolase activity is greatly increased in the presence of bile salts. ... 

In the duodenum, dietary lipids are degraded by pancreatic enzymes triacylglycerol by pancreatic lipase, phospholipids by phospholipase A2 and lysophospholipase, and cholesteryl esters by cholesterol esterase. Enzyme release from the pancreas is controlled by cholecystokinin, produced by cells in the intestinal mucosa. 

Serum cholesterol. Most cholesterol is carried in the blood by low density lipoprotein (LDL, Tables 21-1,21-2), which delivers the cholesteryl esters directly to cells that need cholesterol. Both a 74-kDa cholesteryl ester transfer protein193-1953 and a phospholipid transfer protein196 1963 are also involved in this process. Cholesterol esterases, which release free cholesterol, may act both on lipoproteins and on pancreatic secretions.197-199... 

Cholesterol assay solution. The stock reagent contains pancreatic cholesterol esterase, microbial cholesterol oxidase, horseradish peroxidase, 4-aminoantipyrine, and phenol. Your instructor will reconstitute the stock reagent by addition of water. 

The carboxypeptidases are released from their inactive precursors in the pancreatic juice of animals. The most studied example is bovine carboxypeptidase A, which contains one mole of zinc per protein molecular weight of 34 500. These enzymes cleave the C-terminal amino acid residue from peptides and proteins, when the side-chain of the C-terminal residue is aromatic or branched aliphatic of l configuration. At least the first five residues in the substrate affect the activity of the enzyme. The enzyme also shows esterase activity. Esters and peptides inhibit each other competitively, indicating that the peptidase and esterase sites overlap, even if they are not the same. 

Scriba, G K. E. 1993. Phenytoin-lipid conjugates chemical, plasma esterase-mediated, and pancreatic lipase-mediated hydrolysis vitro. Pharm. Res10 1181-1186. 

Pancreatic cholesterol esterase (3.1.1.3.) aids in transporting cholesterol to the enterocyte. By utilizing a selective and potent cholesterol esterase inhibitor 6-chloro-3-(l-ethyl-2-cyclohexyl)-2-pyrone, the absorption of cholesterol in hamsters could be reduced [71]. Wadkins et al. [72] synthesized novel sulfonamide derivatives, which demonstrated greater than 200-fold selectivity for human intestinal carboxylesterase compared with the human liver carboxylesterase hCEl, and none of them was an inhibitor of human acetylcholinesterase or butyrylcholinester-ase. Maybe these agents can serve as lead compounds for the development of effective, selective carboxylesterase inhibitors for clinical applications. Also the potent P-gp inhibitor verapamil [73] as well as S,S,S-tributylphosphortrithionate (DEF) [74] may exhibit carboxylesterase inhibitory properties. Various other inhibitors of human esterases are listed in Table 5.6. 

Spilburg, C.A., et al. 1995. Identification of a species specific regulatory site in human pancreatic cholesterol esterase. Biochemistry 34 15532. 

---