Interface-mediated inactivation of pancreatic lipase by a water-reactive compound: 2-sulfobenzoic cyclic anhydride

Moulin, A.; Fourneron, Jean‐Dominique; Piéroni, G; Verger, Robert

doi:10.1021/bi00441a029

Cited by 19 publications

(5 citation statements)

References 26 publications

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“…Irreversible inhibitors E600 and PMSF, and THL (reversible inhibitor of pancreatic lipases and reversible in microbial lipases) have been recognized as serine lipase inhibitors since they react with the catalytic serine residue; E600 inhibited 62.80 ± 0.74% and THL only 18.57 ± 2.35% (Table 1). PMSF did not inhibit lipolytic activity under the same conditions, probably because the active site of lipases was accessible only when the enzyme was bound to interfaces (Moulin et al, 1989); similar results were obtained with lipase from Rhizomucor miehei (Fig. S1).…”

Section: Discussionsupporting

confidence: 80%

Cannonball jellyfish digestion: an insight into the lipolytic enzymes of the digestive system

Martínez-Pérez

Rodríguez

Soto

et al. 2020

PeerJ

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The digestive system and metabolism of the cannonball jellyfish Stomolophus sp. 2 are not well-known. The digestion study was critical to explain its ecology and bloom success. Different enzymes are involved in food digestion, which hydrolyze carbohydrates, proteins, and lipids. This study detected lipolytic activity in enzymatic extracts from gastric pouches of Stomolophus sp. 2 collected in the summer of 2013 at Bahía de Kino, Sonora, México (28°47′47″N 111°57′25″W). Lipase/esterase activity showed optimal pH at 11.0 and 50–60 °C with a half-life (t1/2) of 33 min at 55 °C, whereas halotolerance of this activity was recorded from 0-4 M NaCl. Metal ions Ca2+ and Mn2+ did not affect the activity, but Mg2+ decreased it 14.2% ± 3.15, while chelating agents as ethylenediaminetetraacetic acid reduced the activity 8.55% ± 2.13. Inhibition of lipase/esterase activity with tetrahydrolipstatin and paraoxon-ethyl decreased the activity 18.2% ± 2.3, and 62.80% ± 0.74, respectively, whereas phenylmethanesulfonyl fluoride (a protease inhibitor) did not affect it. The enzyme displayed a higher specificity for short-chain triglycerides, but triolein, coconut oil, olive oil, and fish oil were hydrolyzed. For the first time, phospholipase activity from the gastric pouch of Stomolophus sp. 2 was detected using L-α-phosphatidylethanolamine from chicken egg yolk as a substrate. These results suggest that Stomolophus sp. 2 hydrolyze several kinds of lipids, and lipolytic enzymes are active at alkaline pH under different saline conditions, which may be essential to digest different preys.

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Section: Discussionsupporting

confidence: 80%

Cannonball jellyfish digestion: an insight into the lipolytic enzymes of the digestive system

Martínez-Pérez

Rodríguez

Soto

et al. 2020

PeerJ

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“…With this experimental protocol, the binding of E600-modified lipases was found to be comparable to that of native lipases (see Table IV). Furthermore, when DP-PPL was used in the presence of a tributyroylglycerol emulsion, the interface-mediated labeling reaction with [3H]sulfobenzoic cyclic anhydride was still possible (Moulin et al, 1989), which also indicates that DP-PPL was present at the lipid/water interface.…”

Section: Resultsmentioning

confidence: 99%

Inactivation of gastric and pancreatic lipases by diethyl p-nitrophenyl phosphate

Moreau¹,

Moulin²,

Gargouri³

et al. 1991

Biochemistry

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Reacting gastric and pancreatic lipases with mixed diethyl p-nitrophenyl phosphate/bile salt micelles resulted in a stoichiometric inactivation of these enzymes as tested on emulsified tributyroylglycerol and trioleoylglycerol as substrates. Diethyl p-nitrophenyl phosphate treated gastric lipases were also inactive on water-soluble p-nitrophenyl acetate, whereas the modified pancreatic lipase was still able to hydrolyze this water-soluble substrate. The binding of diethyl p-nitrophenyl phosphate modified pancreatic and gastric lipases to tributyroylglycerol/water interface was comparable to that of native lipases. The essential free sulfhydryl group of gastric lipases underwent no chemical changes due to the reaction with micellar diethyl p-nitrophenyl phosphate. All in all, these results indicate that, in both gastric and pancreatic lipases, the essential serine residue which was stoichiometrically labeled by this organophosphorus reagent is involved in catalysis and not in lipid binding.

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“…It was not clear whether a blank sample was derivatized to rule out contamination. Zu and coworkers derivatized fatty alcohol ethoxylate non-ionic surfactants at the 0.05 mmol level using SBA in the presence of 4dimethylaminopyridine (DMAP), followed by diluting the reaction mixture and subjecting it to liquid chromatography (LC)/ESI-MS or flow injection ESI-MS. [5] The yield was reported to be about 95% based on the assumption that the product (SBA-tagged alcohol) and starting alcohol gave the same response by ESI-MS. SBA has also been used to modify proteins [6,7] and industrial polymers. [8] Towards a goal of improving the detection of phenols by MALDI-MS, we have extended the work of others concerning derivatization with SBA, and this manuscript reports our progress.…”

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confidence: 99%

Evaporative derivatization of phenols with 2-sulfobenzoic anhydride for detection by matrix-assisted laser desorption/ionization mass spectrometry

Yao

Wang

Giese

2014

Rapid Commun. Mass Spectrom.

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RATIONALE Phenols are an important class of analytes, for example as bioactive environmental contaminants. Towards a goal of improving their detection by MALDI-TOF-MS or MALDI-TOF/TOF-MS, we studied their derivatization with 2-sulfobenzoic anhydride (SBA). We chose SBA for this purpose since it is commercially available, inexpensive, and forms an anionic derivative. METHODS In selected conditions developed here for phenols, a reaction mixture of one or more of such compounds in acetonitrile containing SBA and 4-dimethylaminopyridine (DMAP) is evaporated to a solid, heated at 60°C for 1 h, redissolved in 50% acetonitrile containing matrix, spotted onto a MALDI target, and subjected to negative ion MALDI-TOF/TOF-MS. RESULTS While conventional (solution-phase) reaction of 4-phenylphenol (model analyte) with SBA and DMAP only gave a 47% yield of SBA-tagged 4-phenylphenol, evaporative derivatization as above gave a 96% yield, and 25 pmol (4.3 ng) of 4-phenylphenol could be detected in this way by MALDI-TOF/TOF-MS at S/N = 260, whereas even 1 nmol of the nonderivatized phenol was not detected in the absence of derivatization. A wide range of responses was observed when a mixture of 15 phenols was derivatized, with the higher responses coming from phenols with a pKa value above 9. Without derivatization, phenols with pKa values below 5 were the most readily detected. CONCLUSION Evaporative derivatization with SBA (a convenient reagent) can improve the detection of phenols with relatively high pKa values (above 9) by negative ion MALDI-TOF-MS, and accomplish this in the absence of post-derivatization reaction cleanup.

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Interface-mediated inactivation of pancreatic lipase by a water-reactive compound: 2-sulfobenzoic cyclic anhydride

Cited by 19 publications

References 26 publications

Cannonball jellyfish digestion: an insight into the lipolytic enzymes of the digestive system

Cannonball jellyfish digestion: an insight into the lipolytic enzymes of the digestive system

Inactivation of gastric and pancreatic lipases by diethyl p-nitrophenyl phosphate

Evaporative derivatization of phenols with 2-sulfobenzoic anhydride for detection by matrix-assisted laser desorption/ionization mass spectrometry

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