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Smaali M. Issam Gargouri Mohamed Legoy Marie Dominique Maugard Thierry Limam Farid Marzouki Nejib 《Applied biochemistry and biotechnology》2004,112(2):63-77
The filamentous fungus Sclerotinia sclerotiorum, grown on a xylose medium, was found to excrete one β-glucosidase (β-glu x). The enzyme was purified to apparent homogeneity
by ammonium sulfate precipitation, gel filtration, anion-exchange chromatography, and high-performance liquid chromatography
(HPLC) gel filtration chromatography. Its molecular mass was estimated to be 130 kDa by HPLC gel filtration and 60 kDa by
sodium dodecyl sulfate polyacrylamide gel electrophoresis, suggesting that β-glu x may be a homodimer. For p-nitrophenyl β-d-glucopyranoside hydrolysis, apparent K
m and V
max values were found to be 0.09 mM and 193 U/mg, respectively, while optimum temperature and pH were 55–60°C and pH 5.0, respectively. β-Glu x was strongly
inhibited by Fe2+ and activated about 35% by Ca2+. β-Glu x possesses strong transglucosylation activity in comparison with commercially available β-glucosidases. The production
rate of total glucooligosaccharides (GOSs) from 30% cellobiose at 50°C and pH 5.0 for 6 h with 0.6 U/mL of enzyme preparation
was 80 g/L. It reached 105 g/L under the same conditions when using cellobiose at 350 g/L (1.023 M). Finally, GOS structure was determined by mass spectrometry and 13C nuclear magnetic resonance spectroscopy. 相似文献
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Issam SM Mohamed G Dominique LM Thierry M Farid L Nejib M 《Applied biochemistry and biotechnology》2004,112(2):63-78
The filamentous fungus Sclerotinia sclerotiorum, grown on a xylose medium, was found to excrete one beta-glucosidase (beta-glu x). The enzyme was purified to apparent homogeneity by ammonium sulfate precipitation, gel filtration, anion-exchange chromatography, and high-performance liquid chromatography (HPLC) gel filtration chromatography. Its molecular mass was estimated to be 130 kDa by HPLC gel filtration and 60 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis, suggesting that beta-glu x may be a homodimer. For p-nitrophenyl beta-d-glucopyranoside hydrolysis, apparent Km and Vmax values were found to be 0.09 mM and 193 U/mg, respectively, while optimum temperature and pH were 55-60 degrees C and pH 5.0, respectively. beta-Glu x was strongly inhibited by Fe2+ and activated about 35% by Ca2+. beta-Glu x possesses strong transglucosylation activity in comparison with commercially available beta-glucosidases. The production rate of total glucooligosaccharides (GOSs) from 30% cellobiose at 50 degrees C and pH 5.0 for 6 h with 0.6 U/mL of enzyme preparation was 80 g/L. It reached 105 g/L under the same conditions when using cellobiose at 350 g/L (1.023 M). Finally, GOS structure was determined by mass spectrometry and 3C nuclear magnetic resonance spectroscopy. 相似文献
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Ines Belhaj-Ben Romdhane Zamen Ben Romdhane Maha Bouzid Ali Gargouri Hafedh Belghith 《Applied biochemistry and biotechnology》2013,171(8):1986-2002
Waste frying oil, which not only harms people’s health but also causes environmental pollution, can be a good alternative to partially substitute petroleum diesel through transesterification reaction. This oil contained 8.8 % of free fatty acids, which cause a problem in a base-catalyzed process. In this study, synthesis of biodiesel was efficiently catalyzed by the covalently immobilized Talaromyces thermophilus lipase and allowed bioconversion yield up to 92 % after 24 h of reaction time. The optimal molar ratio was four to six parts of methanol to one part of oil with a biocatalyst loaded of 25 wt.% of oil. Further, experiments revealed that T. thermophilus lipase, immobilized by a multipoint covalent liaison onto activated chitosan via a short spacer (glutaraldehyde), was sufficiently tolerant to methanol. In fact, using the stepwise addition of methanol, no significant difference was observed from the one-step whole addition at the start of reaction. The batch biodiesel synthesis was performed in a fixed bed reactor with a lipase loaded of 10 g. The bioconversion yield of 98 % was attained after a 5-h reaction time. The bioreactor was operated successfully for almost 150 h without any changes in the initial conversion yield. Most of the chemical and physical properties of the produced biodiesel meet the European and USA standard specifications of biodiesel fuels. 相似文献
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Nabil Smichi Ahmed Fendri Raja Chaabouni Faouzi Ben Rebah Youssef Gargouri Nabil Miled 《Applied biochemistry and biotechnology》2010,162(5):1483-1496
A lipolytic activity was located in the sardine digestive glands (pyloric caeca), from which a sardine digestive lipase (SaDL)
was purified. Pure SaDL has a molecular mass of 43 kDa as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis
analysis. The enzyme was found to be more active on short-chain triacylglycerols than on long-chain ones. SaDL does not present
the interfacial activation phenomenon. Control experiments were performed under the same experimental conditions, with dromedary
and turkey pancreatic lipases and showed a positive interfacial activation phenomenon. Sodium deoxycholate (NaDC) has an inhibitory
effect on the lipase activity. The pure enzyme lost 40% of its activity in presence of 8 mM NaDC. SaDL was found to be mostly
stable at low pH values. Interestingly, no colipase was detected in the sardine pyloric caeca. Analogous results were reported
for the scorpion and the crab digestive systems. This is in line with the idea that colipase might has evolved in mammal animals
simultaneously with the appearance of an exocrine pancreas. No similarity was found between the NH2-terminal amino acid residues of SaDL and those of lipases from the digestive tract of other species. Altogether, these results
suggest that SaDL is a member of a new group of lipases belonging to aquatic species. 相似文献
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Ines Ben Rejeb Lotfi Monser Mohamed Gargouri 《Applied biochemistry and biotechnology》2010,162(6):1536-1546
A new, simple, and original method is described for specific measurement of polyunsaturated fatty acid content in olive oil. This analytical system uses coupled enzymes, lipase and lipoxygenase. The system consists of lipase-catalyzed hydrolysis of triacylglycerol and subsequent lipoxygenation of liberated polyunsaturated fatty acids. The hydroperoxy-fatty acids formed were easily monitored by spectrophotometry at 234 nm. After being optimized, the method was validated in terms of linearity, precision sensitivity, and recovery. Linear calibration graph was obtained in the range 50–500 µg mL?1, with a correlation coefficient higher than 0.921 and a detection limit (S/N?=?3) of 15 µg mL?1. The precision of the method (relative standard deviation) for within and between days is better than 7% and 12%, respectively. The proposed method was successfully applied to the estimation of polyunsaturated fatty acids level in olive oil samples and results obtained were in excellent agreement with those obtained by the classical official method. The proposed method is accurate, simple, cheap, and can be satisfactorily used for routine analysis of edible oils. 相似文献
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Zied Zarai Houda Gharsallah Adnane Hammami Hafedh Mejdoub Sofiane Bezzine Youssef-Talel Gargouri 《Applied biochemistry and biotechnology》2012,168(4):877-886
In order to report pharmacological characterization of marine snail (Hexaplex trunculus) hepatopancreatic phospholipase A2 (mSDPLA2), we have talked for the first time the antimicrobial activity against different pathogenic bacterial strains, anti-chlamydial activity as well as its cytotoxic activity against McCoy cell lines. mSDPLA2, showed a high level of activity towards Gram-positive bacteria as Staphylococcus aureus and Staphylococcus epidermidis. Whereas Gram-negative bacteria, unfortunately, exhibited a higher resistance, mSDPLA2 was also found to have a strong cytotoxic activity, causing significant morphological alterations of the McCoy cell lines surfaces and to be a hinder to the proliferation. Moreover, mSDPLA2 proved to have a very potent anti-chlamydial activity. Over 95?% inhibition of chlamydial inclusions were obtained at a concentration of 10???g/ml of mSDPLA2 after 24?h postinfection. Interestingly, at a concentration of 10???g/ml of mSDPLA2, the proliferation of McCoy cells was not affected. Approximately 50?% inhibition of cell growth was obtained with a concentration of 37???g/mL of mSDPLA2. mSDPLA2 could be considered as an excellent candidate for the development of a new anti-infective agent. This enzyme showed significant antimicrobial activities. 相似文献
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Sawsan Amara Ahmed Fendri Nadia Ben Salem Youssef Gargouri Nabil Miled 《Applied biochemistry and biotechnology》2010,162(4):942-952
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. 相似文献
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Ben Rebah F Smaoui S Frikha F Gargouri Y Miled N 《Applied biochemistry and biotechnology》2008,151(1):71-79
The lipase inhibitory activity of ethanol extracts obtained from some marine algae collected on the Tunisian coast was evaluated.
Caulerpa prolifera extract markedly reduced both dog gastric (DGL) and human pancreatic lipase (HPL) activities. Generally, the inhibition reached
100% after 40 to 60 min of incubation depending on lipase types and on substrates used. Moreover, the inhibitory effect of
C. prolifera extract on lipases appeared to be accelerated by adding bile salts, which likely modified the interface and allowed the inhibitory
compound to inactivate the lipase. The separation of C. prolifera extract by thin-layer chromatography (TLC) resulted in eight fractions showing efficient inhibition rate against DGL, compared
to the crude extract. In the case of HPL, TLC fractionation reduced the inhibitory rates, suggesting that the effect of algal
extract on lipases may be caused by a synergetic action of several compounds within the extract. High-performance liquid chromatograph
separation resulted in isolation of a major compound displaying high inhibition capacity of HPL activity. Caulerpa prolifera extract may therefore be useful in developing antiobesity drugs. 相似文献