ABSTRACT
Interfacial esterases are useful enzymes in bioconversion and racemic mixture resolution processes. Marine invertebrates are few explored potential sources of these proteins. In this work, aqueous extracts of 41 species of marine invertebrates were screened for esterase, lipase, and phospholipase A activities, being all positive. Five extracts (Stichodactyla helianthus, Condylactis gigantea, Stylocheilus longicauda, Zoanthus pulchellus, and Plexaura homomalla) were selected for their activity values and immobilized on Octyl-Sepharose CL 4B support by interfacial adsorption. The selectivity of this immobilization method for interfacial esterases was evidenced by immobilization percentages ≥ 94% in almost all cases for lipase and phospholipase A activities. Six pharmaceutical-relevant esters (phenylethyl butyrate, ethyl-2-hydroxy-4-phenyl-butanoate, 2-oxyranylmethyl acetate (glycidol acetate), 7-aminocephalosporanic acid, methyl-prostaglandin F2α, and methyl-6-metoxy-α-methyl-2-naphtalen-acetate -naproxen methyl ester-) were bioconverted by at least three of these biocatalysts, with the lowest conversion percentage of 24%. In addition, three biocatalysts were used in the racemic mixture resolution of three previous compounds. The S. helianthus-derived biocatalyst showed the highest enantiomeric ratios for glycidol acetate (2.67, (S)-selective) and naproxen methyl ester (8.32, (R)-selective), and the immobilized extract of S. longicauda was the most resolutive toward the ethyl-2-hydroxy-4-phenyl-butanoate (8.13, (S)-selective). These results indicate the relevance of such marine interfacial esterases as immobilized biocatalysts for the pharmaceutical industry.
Subject(s)
Aquatic Organisms/enzymology , Biocatalysis , Enzymes, Immobilized/chemistry , Enzymes, Immobilized/metabolism , Esterases/chemistry , Esterases/metabolism , Invertebrates/enzymology , Animals , Esters/chemistry , Esters/metabolism , Lipase/metabolism , Phospholipases/metabolism , Stereoisomerism , Substrate Specificity , Water/chemistryABSTRACT
In the present study, we report the isolation and characterization of seabream Sparus aurata pyloric caeca-duodenal lipase. Optimum activity was found at pH 8.5 and salinity of 50 mM NaCl. Lipase activity was sensitive to divalent ions, and extreme pH values (4, 5, and 12), being more stable at alkaline than acid pH. Optimum temperature was found at 50°C, but lipase was stable at temperatures below 40°C. Lipase has a bile salt sodium taurocholate requirement for increased activity. Gradient PAGE electrophoresis revealed the presence of four isoforms with apparent molecular masses of 34, 50, 68, and 84 KDa, respectively. Pyloric-duodenal lipase was able to hydrolyze emulsified alimentary oils. Results confirm the presence of true lipases in Sparus aurata digestive tract.
Subject(s)
Duodenum/enzymology , Gastric Mucosa/enzymology , Lipase/metabolism , Sea Bream/physiology , Animals , Cations, Divalent/pharmacology , Enzyme Activation/drug effects , Enzyme Stability , Hydrogen-Ion Concentration , Lipase/isolation & purification , Olive Oil , Plant Oils/metabolism , Protein Isoforms , Sodium Chloride/pharmacology , Taurocholic Acid/pharmacology , TemperatureABSTRACT
Investigation of esterase-lipase activity in the digestive gland of redclaw crayfish Cherax quadricarinatus showed that the optimum enzyme activity occurred between 35 and 40 degrees C, with 100 mM NaCl at pH 8.5. Heavy metals completely inhibited and calcium ions partially inhibited enzyme activity. The enzyme activity diminished as the length of the fatty acid chain of substrates increased. Molecular masses for four isozymes were 43, 46, 63 and 118 kDa, respectively, as determined by PAGE.
