Crab digestive lipase acting at high temperature: Purification and biochemical characterization

Chérif, Slim; Fendri, Ahmed; Miled, Nabil; Trabelsi, Heykel; Mejdoub, Hafedh; Gargouri, Youssef

doi:10.1016/j.biochi.2007.02.005

Cited by 52 publications

(38 citation statements)

References 28 publications

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“…The pH value for maximal lipase activity and the high level of activity throughout a wide range of pH (5.4-8.5) (Fig. 1A) is in agreement with that found for this enzyme in hepatopancreas of the euryhaline crab N. granulata (Michiels et al 2011) and of the marine green crab Carcinus mediterranus (Cherif et al 2007, Cherif and Gargouri 2009, Smichi et al 2012. The high lipase activity at 37°C in hepatopancreas of C. angulatus (Fig.…”

Section: Discussionsupporting

confidence: 87%

Effect of environmental salinity and dopamine injections on key digestive enzymes in hepatopancreas of the euryhaline crab <i>Cyrtograpsus angulatus</i> (Decapoda: Brachyura: Varunidae)

Michiels¹,

Valle²,

Mañanes³

2012

Sci. Mar.

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SUMMARY:We studied the occurrence and characteristics of lipase activity and the response of lipase and proteolytic activity to salinity and dopamine injections in hepatopancreas of the euryhaline crab (Cyrtograpsus angulatus). Lipase activity was maximal at pH 8.5; it exhibited Michaelis-Menten kinetics (apparent K m =0.019 mM), was higher at 37°C and appeared to be cold tolerant, being also high at 4°C. In 10 psu (hyper-regulation conditions), lipase and proteolytic activity were about 3 and 5 times higher, respectively, than in 35 psu (osmoconformation). In 40 psu (hypo-regulation), lipase activity was about three times higher than in 35 psu, while proteolytic activity was similar. Lipase activity was inhibited in vivo by 10 -4 M dopamine in 35 psu but not in 10 or 40 psu. Proteolytic activity was not affected by 10 -4 M dopamine. The differential responses of lipase and proteolytic activity to salinity and dopamine suggest the occurrence of distinct digestive adjustments and mechanisms of regulation upon osmoregulatory conditions. This study contributes to a better understanding of the complexity of the biochemical adaptations to salinity in euryhaline crabs. The fact that higher digestive enzyme activities could be associated with a differential digestive capacity potentially leading to enhanced availability of energy substrates is discussed.Keywords: Cyrtograpsus angulatus, euryhaline crabs, salinity, hepatopancreas, digestive enzymes, dopamine, Mar Chiquita lagoon. RESUMEN: EfEcto dE la salinidad ambiEntal y dE la inyEcción dE dopamina sobrE Enzimas digEstivas clavE En hEpatopáncrEas dEl cangrEjo EurihalinoCyrtograpsus angulatus (dEcapoda: brachyura: varunidaE). -Se estudió la existencia y características bioquímicas de actividad de lipasa y la respuesta a salinidad y a dopamina de actividad de lipasa y proteolítica total en el hepatopáncreas del cangrejo eurihalino Cyrtograpsus angulatus. La actividad de lipasa fue máxima a pH 8,5; mostró una cinética michaeliana (K m =0.019 mM); fue mayor a 37°C, y resistente a baja temperatura (4°C). En 10 psu (condición de hiperregulación), las actividades de lipasa y proteolítica fueron 3 y 5 veces mayores, respectivamente, que en 35 psu (condición de osmoconformación). En 40 psu (condición de hiporregulación), la actividad de lipasa fue de alrededor 3 veces mayor que en individuos aclimatados a 35 psu, mientras que la actividad proteolítica fue similar. La actividad de lipasa fue inhibida in vivo por dopamina 10 -4 M en 35 psu pero no fue afectada en 10 o 40 psu. Dopamina 10 -4 M no afectó la actividad proteolítica. Las respuestas de las actividades de lipasa y proteolítica a salinidad y a dopamina permiten sugerir la existencia de ajustes digestivos y mecanismos de regulación diferenciales en relación al estado osmorregulatorio. Los resultados constituyen un aporte relevante al conocimiento sobre las complejas adaptaciones a nivel bioquímico a la salinidad en cangrejos eurihalinos.

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Section: Discussionsupporting

confidence: 87%

Effect of environmental salinity and dopamine injections on key digestive enzymes in hepatopancreas of the euryhaline crab <i>Cyrtograpsus angulatus</i> (Decapoda: Brachyura: Varunidae)

Michiels¹,

Valle²,

Mañanes³

2012

Sci. Mar.

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“…Our data (not shown) indicated that unlike vertebrates, no detectable yields of bile salts were found in the snail digestive glands. This result is in line with that found in the case of other invertebrates, such as scorpion [6] and crab [7].…”

Section: Investigation Of Colipase and Bile Salts Levels In The Snailsupporting

confidence: 91%

“…Lipase activity decreased significantly above pH 9.0, and only 5% of the residual activity was measured at pH9.5. The optimum pH for SnDL was similar to that reported for the crab lipase [7] and was lower than that of the scorpion lipase (pH9) [6].…”

Section: Effect Of Ph On Sndl Activitymentioning

confidence: 54%

“…Since the detection of the presence of lipase activity in the gastric juice of some crustaceans (Homarus americanus) and mollusks (Aplysia californica) [4,5], a number of investigations on invertebrate lipases have been reported. Recently, lipases were isolated from digestive glands of lower animals, such as the scorpion [6] and the crab [7]. The purified enzymes showed no similarity with any known lipases.…”

Section: Introductionmentioning

confidence: 98%

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Snail Hepatopancreatic Lipase: A New Member of Invertebrates Lipases' Group

Amara¹,

Fendri²,

Salem³

et al. 2009

Appl Biochem Biotechnol

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Higher animal's lipases are well characterized; however, much less is known about lipases from mollusks. A lipolytic activity was located in the land snail (Eobania vermiculata) digestive glands (hepatopancreas), from which a snail digestive lipase (SnDL) was purified. Pure SnDL has a molecular mass of 60 kDa; it does not present the interfacial activation phenomenon. It was found to be more active on short-chain triacylglycerols than on long-chain triacylglycerols. The NH2-terminal sequence of the SnDL shows 66% of identity with the 17 NH2-terminal amino acids of a putative lipase from sea urchin (Strongylocentrotus purpuratus). No sequence identity was found with known lipases. Interestingly, neither colipase nor bile salts were detected in the snail hepatopancreas. This suggests that colipase evolved in vertebrates simultaneously with the appearance of an exocrine pancreas and a true liver which produces bile salts. Altogether, these results suggest that SnDL is a member of a new group of digestive lipases belonging to invertebrates.

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“…8). Crab digestive lipase (Cherif et al 2007) was also stable in this pH range, but in addition had significantly higher residual activities at pH 8-10 than the two lipases in our study. A lipase from the hepatopancreas of oil sardine was stable between pH 5 and 9.5 (Mukundan et al 1985) and intestinal lipase from Nile tilapia was stable between pH 6.5 and 8.5 (Taniguchi et al 2001).…”

Section: Acyl-chain Specificitymentioning

confidence: 39%

Purification and properties of digestive lipases from Chinook salmon (Oncorhynchus tshawytscha) and New Zealand hoki (Macruronus novaezelandiae)

Kurtovic

Marshall

Zhao

et al. 2010

Fish Physiol Biochem

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Lipases were purified from delipidated pyloric ceca powder of two New Zealand-sourced fish, Chinook salmon (Oncorhynchus tshawytscha) and hoki (Macruronus novaezelandiae), by fractional precipitation with polyethylene glycol 1000, followed by affinity chromatography using cholate-Affi-Gel 102, and gel filtration on Sephacryl S-300 HR. For the first time, in-polyacrylamide gel activity of purified fish lipases against 4-methylumbelliferyl butyrate has been demonstrated. Calcium ions and sodium cholate were absolutely necessary both for lipase stability in the gel and for optimum activity against caprate and palmitate esters of p-nitrophenol. A single protein band was present in native polyacrylamide gels for both salmon and hoki final enzyme preparations. Under denaturing conditions, electrophoretic analysis revealed two bands of 79.6 and 54.9 kDa for salmon lipase. It is proposed that these bands correspond to an uncleaved and a final form of the enzyme. One band of 44.6 kDa was seen for hoki lipase. pI values of 5.8±0.1 and 5.7±0.1 were obtained for the two salmon lipase forms. The hoki lipase had a pI of 5.8±0.1. Both lipases had the highest activity at 35°C, were thermally labile, had a pH optimum of 8-8.5, and were more acid stable compared to other fish lipases studied to date. Both enzymes were inhibited by the organophosphate paraoxon. Chinook salmon and hoki lipases showed good stability in several water-immiscible solvents. The enzymes had very similar amino acid composition to mammalian carboxyl ester lipases and one other fish digestive lipase. The salmon enzyme was an overall better catalyst based on its higher turnover number (3.7±0.3 vs. 0.71±0.05 s(-1) for the hoki enzyme) and lower activation energy (2.0±0.4 vs. 7.6±0.8 kcal/mol for the hoki enzyme) for the hydrolysis of p-nitrophenyl caprate. The salmon and hoki enzymes are homologous with mammalian carboxyl ester lipases.

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Crab digestive lipase acting at high temperature: Purification and biochemical characterization

Cited by 52 publications

References 28 publications

Effect of environmental salinity and dopamine injections on key digestive enzymes in hepatopancreas of the euryhaline crab <i>Cyrtograpsus angulatus</i> (Decapoda: Brachyura: Varunidae)

Effect of environmental salinity and dopamine injections on key digestive enzymes in hepatopancreas of the euryhaline crab <i>Cyrtograpsus angulatus</i> (Decapoda: Brachyura: Varunidae)

Snail Hepatopancreatic Lipase: A New Member of Invertebrates Lipases' Group

Purification and properties of digestive lipases from Chinook salmon (Oncorhynchus tshawytscha) and New Zealand hoki (Macruronus novaezelandiae)

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