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1.
A detailed kinetic study was carried out to investigate the porcine pancreatic lipase-catalysed esterification reactions of p-cresol–acetic acid and lactic acid–stearic acid. The kinetic data were in agreement with a Ping Pong Bi–Bi mechanism being followed by the enzyme, where inhibition is indicated in the presence of p-cresol and lactic acid in the respective reactions. Mathematical analyses of experimentally observed initial rates yielded various kinetic parameters, K m(p-cresol) = 0.1, K m(acetic acid) = 0.54, K m(lactic acid) = 0.059 M, K m(stearic acid) = 0.04 M, V max(p-cresol–acetic acid) = 13.2(h–1), V max(lactic acid–stearic acid) = 0.00163 M/h, K i(p-cresol) = 0.59 and K i(lactic acid) = 0.079 M. The K m and K i values of p-cresol and lactic acid observed in the respective reactions showed both the competitive nature of binding between the substrates p-cresol and acetic acid on the one hand and lactic acid and stearic acid on the other and the inhibitory nature of p-cresol and lactic acid.  相似文献   

2.
(R)-2-Phenylpropanoic acid was synthesized from the racemic acid through an isomerization reaction involving resting cells of Nocardia diaphanozonaria JCM3208. The isomerization activity of the cells was enhanced 25-fold by adding 5.5 mM racemic 2-phenylpropanoic acid to the culture medium. When 5 mM racemic 2-phenylpropanoic acid was included in the reaction mixture (4 ml) containing resting cells (100 mg dry cell wt) in 25 mM K2HPO4/KH2PO4 buffer (pH 7.0) at 30 °C for 8 h, 4.56 mM (R)-2-phenylpropanoic acid (95.8% e.e.) was formed with a 91% molar conversion yield.  相似文献   

3.
Two new natural triterpenes, lantaninilic acid and lantoic acid, along with the known triterpenes lantadene A, and oleanolic, ursolic, betulinic, lantanolic, and camaric acid, were obtained from the aerial parts of Lantana camara through bioassay‐guided isolation, monitoring the in vitro antileishmanial activity against promastigotes of Leishmania major. Oleanolic acid ( 3 ), ursolic acid ( 4 ), lantadene A ( 5 ), and lantanilic acid ( 7 ) showed significant leishmanicidal activities with IC50 values of 53.0, 12.4, 20.4, and 21.3 μM , respectively. The IC50 value of ursolic acid ( 4 ; 12.4 μM ) was found to be comparable with that of the standard drugs, pentamidine (IC50 15.0 μM ) and amphotericin B (IC50 0.31 μM ). The in vitro activities of L. camara and its constituents against promastigotes of Leishmania major are reported here for the first time.  相似文献   

4.
Enrichment of n-3 polyunsaturated fatty acids from sardine cannery effluents upon enzymatic esterification by Lipozyme® was optimized in batch and in continuous processes. In these processes, the yield of docosahexaenoic acid (DHA) (Y1) and the yield of eicosapentaenoic acid (EPA) (Y2), in the residual acid fraction were maximized. In batch, a two-factor Doehlert design was used to study the effects of temperature and ratio of fatty acid to alcohol. Second order polynomial regression models for Y1 and Y2 were postulated to generate response surfaces. After esterification the fraction of fatty acid was enriched to 70% with DHA or to 44% with EPA. In a continuous process, a three-factor central composite design was employed to study the effects of temperature, ratio of fatty acid to alcohol and flow rate. Second order polynomial regression models for Y1 and Y2 were used to generate response surfaces. After esterification, a quantity of DHA close to 30% and 17% of EPA.  相似文献   

5.
The effect of organic solvents on carboxypeptidase Y (a serine carboxypeptidase from yeast)-catalyzed hydrolysis of amino acid ester and peptide synthesis from N-acyl amino acid ester and amino acid amide was investigated.

The Km value of ester hydrolysis increased with an increase in the solvent content. Dioxane was the most effective and dimethyl sulfoxide (DMSO) the least, whilst Kcat showed a tendency to increase slightly in N, N-dimethylformamide (DMF) and DMSO. For dioxane and acetonitrile (MeCN) a maximum was observed.

In peptide formation from Fua-Phe-OEt and Gly-NH2, dioxane and MeCN supported high product yield at molar fractions smaller than ca. 0.05 but the yield decreased significantly at higher fractions, although a relatively constant selectivity (ratio of the peptide bond formed to the ester consumed) was maintained. DMSO gave rather low peptide yields and selectivity even at lower molar fractions. DMF showed an intermediate tendency.

An apparent saturation parameter of the amine component was evaluated and the dissociation constant of a complex between acyl-enzyme and amino acid amide (Kn), as well as the rate constant of aminolysis exerted by the amino acid amide bound correctly on the enzyme (Kn), was calculated by initial rate analysis of peptide formation. In contrast to Km values, Kn decreased with increasing concentrations of organic cosolvent. while a suppressive effect was observed (except for DMSO) on the Kn parameter.

Effects of the solvent practically immiscible in water was also studied by use of the enzyme physically “immobilized” on glass beads.  相似文献   

6.
Effect of water activity (a w) adjusted with NaCl or glycerol on the growth and metabolism ofLactobacillus helveticus var.pragensis at 40°C was demonstrated by growth curves (generation time) and changes in the degree of acidity of a milk medium expressed as lactic acid content. NaCl-regulateda w of 0.970 inhibited growth and acid production completely. The same, glycerol-regulateda w of 0.970 increased the generation time only from 33 min (a w=0.994) to 67 min which was less than the generation time at the highera w of 0.982 adjusted with NaCl (103 min). Glycerol-regulateda w of 0.970 did not decrease the acid production (2.6% lactic acid). Ata w of 0.951, the acid production was decreased by 39% compared with the values found in milk media with the original, unadjusteda w of 0.994, after the same time of incubation. Media witha w adjusted to the same values with NaCl or glycerol do not influence the growth and acid production ofL. helveticus in the same way. In contrast to glycerol, NaCl has a strong effect.  相似文献   

7.
The ability of ascorbic acid (Vitamin C) to modulate the genotoxic action of several mutagens was investigated in the wing spot test of Drosophila melanogaster. In this assay, 3-day-old transheterozygous larvae for the multiple wing hairs (mwh, 3-0.3) and flare (flr, 3-38.8) genes were treated with three reference mutagenic compounds, namely cobalt chloride (CoCl2), 4-nitroquinoline 1-oxide (4-NQO) and potassium dichromate (K2Cr2O7). The results obtained show that the three reference mutagens tested were clearly genotoxic in the Drosophila wing somatic mutation and recombination test (SMART). None of the three concentrations tested of ascorbic acid (25, 75 and 250 mM) induced significant increases in the frequency of the mutant clones recorded. When co-treatment experiments with ascorbic acid were carried out, different results were found. Thus, ascorbic acid was effective in reducing the genotoxicity of K2Cr2O7 virtually to the control level; on the contrary, it did not show any antigenotoxic effect on the genotoxicity of 4-NQO. Finally, co-treatments with CoCl2 and ascorbic acid show a significant increase in the frequency of mutant clones over the values obtained with CoCl2 alone.  相似文献   

8.
This study examines the effect of 1,25-dihydroxyvitamin D3 [1,25(OH)2D3], 24,25-dihydroxyvitamin D3 [24,25(OH)2D3], two vitamin D analogues (KH 1060 and EB 1089, which are 20-epi-22-oxa and 22,24-diene-analogues, respectively), 9-cis retinoic acid and all-trans retinoic acid on proliferation of SH-SY5Y human neuroblastoma cells, after treatment for 7 days. Cell number did not change when the cells were incubated with 1, 10 or 100 nM 1,25(OH)2D3 or its derivatives, but significantly decreased in the presence of the two retinoids (0.001–10 μM final concentration). A synergistic inhibition was observed, when SH-SY5Y cells were treated combining 0.1 μM 9-cis retinoic acid and 10 nM 1,25(OH)2D3 or 10 nM KH 1060, and 1 μM 9-cis retinoic acid and 10 nM 1,25(OH)2D3 or 10 nM EB 1089. Acetylcholinesterase activity showed a significant increase, in comparison with controls, after treatment of the cells for 7 days with 0.1 or 1 μM 9-cis retinoic acid, alone or combined with 10 nM 1,25(OH)2D3 or 10 nM KH 1060 or 10 nM EB 1089. This increase was synergistic, combining 1 μM 9-cis retinoic acid and 10 nM 1,25(OH)2D3 or EB 1089. The levels of the c-myc encoded protein remarkably decreased after treatment of SH-SY5Y cells for 1, 3, 7 days with 0.1 and 1 μM 9-cis retinoic acid, alone or combined with 10 nM 1,25(OH)2D3 or 10 nM KH 1060 or 10 nM EB 1089. In particular, the association of 1 μM 9-cis retinoic acid and 10 nM 1,25(OH)2D3 or 10 nM EB 1089 resulted in a synergistic c-myc inhibition, in comparison with that obtained in the presence of the retinoid alone. These findings may have therapeutic implications in human neuroblastoma.  相似文献   

9.
Summary. Branched-chain amino acid aminotransferase was purified by several column chromatographies from Helicobacter pylori NCTC 11637, and the N-terminal amino acid sequence was analyzed. The enzyme gene was sequenced based on a putative branched-chain amino acid aminotransferase gene, ilvE of H. pylori 26695, and the whole amino acid sequence was deduced from the nucleotide sequence. The enzyme existed in a homodimer with a calculated subunit molecular weight (MW) of 37,539 and an isoelectric point (pI) of 6.47. The enzyme showed high affinity to 2-oxoglutarate (K m = 0.085 mM) and L-isoleucine (K m = 0.34 mM), and V max was 27.3 μmol/min/mg. The best substrate was found to be L-isoleucine followed by L-leucine and L-valine. No activity was shown toward the D-enantiomers of these amino acids. The optimal pH and temperature were pH 8.0 and 37 °C, respectively.  相似文献   

10.
The fatty acid composition of the fruit oils or seed oils of Pittosporaceae (eight genera, 10 species), Araliaceae (two species), Simarubaceae (three species), and of one umbelliferous and one rutaceous species were determined by gas chromatography, argentation TLC and ozonolysis. In the Pittosporaceae, in which the major C18 fatty acid of all species was either oleic acid (18:1, 9c) or linoleic acid (18:2, 9c, 12c), large amounts of C20 and C22 fatty acids seem to occur regularly. Petroselinic (18:1, 6c) and tariric (18:1, 6a) acids were absent. However, petroselinic acid was the major fatty acid in the Araliaceae and Umbelliferae. In these two families only small amounts of C20 and C22 acids were detected and tariric acid was absent. The Rutales contained relatively high amounts of trans-octadecenoic acids (18:1, 9t). Tariric acid was the major fatty acid in the two species of Picramnia (Simarubraceae), which also contained small amounts of petroselinic acid. The major fatty acids in Ailanthus glandulosa (Simarubaceae) and Phellodendron amurense (Rutaceae) were linoleic or linolenic acid (18:3, 9c, 12c, 15c); these species contained neither tariric nor petroselinic acid and the levels of C20 and C22 fatty acids were low. The appearance of schizogenous resin canals and polyacetylenes and the absence of iridoids and petroselinic acid allows the Pittosporaceae to be separated from the Rutales and Araliales and to be placed in an independent order, the Pittosporales. Arguments for a rather close relationship of the Pittosporales to the Araliales and Cornales (including the Escalloniaceae) are presented.  相似文献   

11.
A single chiral cyclic α,α‐disubstituted amino acid, (3S,4S)‐1‐amino‐(3,4‐dimethoxy)cyclopentanecarboxylic acid [(S,S)‐Ac5cdOM], was placed at the N‐terminal or C‐terminal positions of achiral α‐aminoisobutyric acid (Aib) peptide segments. The IR and 1H NMR spectra indicated that the dominant conformations of two peptides Cbz‐[(S,S)‐Ac5cdOM]‐(Aib)4‐OEt ( 1) and Cbz‐(Aib)4‐[(S,S)‐Ac5cdOM]‐OMe (2) in solution were helical structures. X‐ray crystallographic analysis of 1 and 2 revealed that a left‐handed (M) 310‐helical structure was present in 1 and that a right‐handed (P) 310‐helical structure was present in 2 in their crystalline states. Copyright © 2010 European Peptide Society and John Wiley & Sons, Ltd.  相似文献   

12.
This study was undertaken to investigate: (1) the effects of both deficiencies and excesses in essential amino acids (EAAs) from an estimated optimum dietary EAA pattern on nitrogen (N) utilization and excretion of rainbow trout Oncorhynchus mykiss, (2) the effects of dietary digestible protein (PD): digestible energy (ED) ratio (PD:ED) on N utilization and excretion of O. mykiss and (3) the potential interaction of these two factors. A 3 × 3 factorial experiment was conducted, with the two factors EAA pattern and PD:ED ratio. The three levels of EAA pattern were: (1) optimum EAA pattern, (2) 60% deficiencies in the three amino acids arginine, histidine and lysine, and (3) 60% excesses in the three amino acids arginine, histidine and leucine. The three levels of PD:ED ratio were 18, 21 and 24 g MJ?1. Amino acid deficiencies from an optimum amino acid pattern caused reductions in mean N retention of 29 to 37%, with the greatest reduction associated with the lowest PD:ED ratio, and similar substantial increases in total N and ammonia‐N excretion at all of the dietary PD:ED ratios investigated. Amino acid excesses, however, did not negatively affect N retention or excretion. Increasing PD:ED ratio was associated with decreasing N retention and increasing N excretion over the range of dietary protein and lipid levels tested. Results of this study showed that a diet with optimum dietary amino acid pattern and lowest PD:ED ratio produced the highest N retention (47% of ingested N) and the lowest total N and ammonia‐N excretion of O. mykiss.  相似文献   

13.
The food pathogen Bacillus cereus is likely to encounter acidic environments (i) in food when organic acids are added for preservation purposes, and (ii) during the stomachal transit of aliments. In order to characterise the acid stress response of B. cereus ATCC14579, cells were grown in chemostat at different pH values (pHo from 9.0 to 5.5) and different growth rates (μ from 0.1 to 0.8 h−1), and were submitted to acid shock at pH 4.0. Cells grown at low pHo were adapted to acid media and induced a significant acid tolerance response (ATR). The ATR induced was modulated by both pHo and μ, and the μ effect was more marked at pHo 5.5. Intracellular pH (pHi) was affected by both pHo and μ. At a pHo above 6, the pHi decreased with the decrease of pHo and the increase of μ. At pHo 5.5, pHi was higher compared to pHo 6.0, suggesting that mechanisms of pHi homeostasis were induced. The acid survival of B. cereus required protein neo-synthesis and the capacity of cells to maintain their pHi and ΔpH (pHi - pHo). Haemolysin BL and non-haemolytic enterotoxin production were both influenced by pHo and μ.  相似文献   

14.
Summary Aspergillus terreus NRRL 1960 was grown on porous disks rotating intermittently in and out of the liquid phase. This immobilized fungal cell bioreactor was used to produce itaconic acid from glucose in a continuous operation. The effect of temperature, pH, disk rotation speed, and feed rate on the itaconic acid concentration and volumetric productivity were studied. The highest itaconic acid concentration and volumetric productivity obtained were 18.2 g/l and 0.73 g/l·h, respectively, under the following conditions: temperature at 36°C, pH 3.0, disk rotation speed at 8 rpm, and feed rate at 60 ml/h. These results are better than those by conventional fermentation or by other immobilized method.Nomenclature F feed rate (l/h) - K 1s saturation constant for immobilized cells (g/l) - K 2s saturation constant for suspended cells (g/l) - M 1 increased mass of immobilized cells (g) - M 2 total mass of immobilized cells (g) - P concentration of itaconic acid (g/l) - S substrate concentration in and out of the reactor (g/l) - S 0 substrate concentration in the feed (g/l) - V liquid volume of the reactor (1) - X concentration of the suspended cells (g/l) - Y 1 apparent yield of the immobilized cells (g cells/g substrate) - Y 2 apparent yield of the suspended cells (g cell/g substrate) - Y 3 apparent yield of itaconic acid (g itaconic acid/g substrate) - m 1 maintenance and by-products coefficient of the immobilized cells (g substrate/g cell·h) - m 2 maintenance and by-products coefficient of the suspended cells (g substrate/g cell·h) - µ1max maximum specific growth rate of the immobilized cells (h-1) - µ2max maximum specific growth rate of the suspended cells (h-1)  相似文献   

15.
This study presents the effects of citric acid and extracellular pH (pHe) on the intracellular pH (pHi) of wild-type and citrate negative variants (cit) Leuconostoc mesenteroides subsp. mesenteroides (Ln. mesenteroides M) and Lactococcus lactis subsp. lactis bv. diacetylactis (L. lactis LD). A recent method using a pH-sensitive fluorescent indicator carboxyfluorescein succinimidyl ester (cFSE) was adapted to measure the pHi of these two lactic acid bacteria in resting cells. Energized cells with 10 mM lactose of Ln. mesenteroides M and L. lactis LD modified their pH gradient (ΔpH) in the same manner; when the pHe was decreased from 7 to 4, the pHi decreased from 7 to about 5. The adjunction of 10 mM citric acid had no effect on the pHi of wild-type and cit(−) variant of L. lactis LD, nor on the pHi of Ln. mesenteroides cit(−) variant. Nevertheless, in Ln. mesenteroides M wild-type, citric acid utilization increased the pHi, which was maintained at about 6.5–7.0 when the pHe was decreased from 7 to 4. It could be concluded that citric acid allows the maintenance of pH homeostasis in Leuconostoc mesenteroides. Received: 7 March 1997 / Accepted: 14 April 1997  相似文献   

16.
Phosphatase activities were measured in preparations of vacuoles isolated from storage roots of red beet (Beta vulgaris L.). The vacuoles possessed both acid phosphatase and ATPase activities which could be distinguished by their susceptibility to inhibition by low concentrations of ammonium molybdate [(NH4)6Mo7O24·4H2O]. The acid phosphatase was completely inhibited by 100 M ammonium molybdate but the ATPase was unaffected. The acid phosphatase was a soluble enzyme which hydrolysed a large number of phosphate esters and had a pH optimum of 5.5. In contrast, the ATPase was partially membrane-bound, had a pH optimum of 8.0 and hydrolysed ATP preferentially, although it was also active agianst PPi, GTP and GDP. At pH 8.0 both the ATPase and PPase activities were Mg2+-dependent and were further stimulated by KCl. The ATPase and PPase activities at pH 8.0 may be different enzymes. The recovery and purification of the ATPase during vacuole isolation were determined. The results indicate that the Mg2+-dependent, KCl-stimulated ATPase activity is not exclusively associated with vacuoles.Abbreviations BSA bovine serum albumen - MES 2-(N-Morpholino)ethanesulphonic acid - MOPS 3-(N-Morpholino)propanesulphonic acid - Na2EDTA ethylenediaminetetra-acetic acid, disodium salt - Pi inorganic phosphate - PPi inorganic pyrophosphate - PPase inorganic pyrophosphatase - TCA trichloroacetic acid - TES N-tris(hydroxymethyl)methyl-2-amino-ethanesulphonic acid - Tris tris(hydroxymethyl)methylamine  相似文献   

17.
Strain MR-12 which was derived from Candida cloacae M-l as a mutant unable to assimilate n-alkane showed marked increase in dicarboxylic acid (DC) productivity from n-alkane.

Resting cells of strain MR-12 produced 42.7g/liter of n-tetradecane 1,14-dicarboxylic acid (DC-16) from n-hexadecane (n-C16) after 72 hr’ incubation. DC degradation activities of strain M-1 and MR-12 were found to be markedly reduced and their activities against DC-16 decreased to 40% and 10% of that of the parent strain, respectively.

Strain M-1 and MR-12 produced DC from the various oxidized derivatives of n-alkane such as alcohol, diol, aldehyde, fatty acid and methyl- or ethylester of fatty acid other than n-alkane.

The carbon balance in n-C16 oxidation was determined by using resting cells of strain MR-12 and about 60% of utilized carbon was recovered as DC-16 and about 40% was recovered as CO2.  相似文献   

18.
A number of polytripeptides related to collagen, namely, (Gly-Pro-Pro)n, (Gly-Pro-Hyp)n, (Gly-Hyp-Hyp)n, (Gly-Pro-Ala)n, (Gly-Pro-Leu)n, (Gly-Pro-Gly)n,(Gly-Ala-Pro)n, (Gly-Ala-Hyp)n, (Ala-Pro-Pro)n, and (Ala-Hyp-Hyp)n were investigated by the method of ir spectroscopy and hydrogen-deuterium kinetics. Strength and order of interpeptide hydrogen bonds of the polytripeptides in a triple-helical conformation were found to depend on the amino acid composition and residue sequence in the triplets. Correlation of X-ray diffraction and spectroscopic data for (Gly-Pro-Hyp)n showed that the increase of the helix parameter in the process of dehydration is accompanied with the weakening of interpeptide hydrogen bonds. Influences of bound water on the length and order of interchain hydrogen bonding was also examined. It was shown that the incorporation of water molecules into the triple helix depends on the amino acid composition and residue sequence. Synthetic models and native collagens were compared.  相似文献   

19.
Metabolism of veratric acid and other aromatic compounds has been studied in two strains of Pycnoporus cinnabarinus. In non-agitated cultures which contained cellulose as an additional carbon source, veratric acid was demeth(ox)ylated to vanillic acid which accumulated in the medium. Under these conditions, 14CO2 evolution from [4-O14CH3]-veratric acid preceded that from [3-O14CH3]-veratric acid in the case of both strains. 14CO2 evolution was markedly accelerated and increased when 100% oxygen was employed instead of air. Oxygen had not so strong effect on the decarboxylation of 14COOH-labelled vanillic and p-hydroxybenzoic acid but it did increase decarboxylation of 14COOH-labelled veratric acid, indicating the effect of oxygen on the preceding demeth(ox)ylation. There were indications, for example rapid demethylation of veratric acid in early stages of growth when apparent phenol oxidase (laccase) activity was zero, for an existence of a separate demethylase enzyme. However, the participation of phenol oxidases in demeth(ox)ylation cannot be ruled out. Degradation pattern of vanillic acid was basically similar in P. cinnabarinus compared to Sporotrichum pulverulentum (Phanerochaete chrysosporium). Also the effect of carbon source was similar: cellulose as a carbon source enhanced degradation of vanillic acid through methoxyhydroquinone whereas in glucose medium, vanillic acid was reduced to the respective aldehyde and alcohol.Non-standard abbreviations CBQ cellobiose: quinone oxidoreductase - MHQ methoxyhydroquinone  相似文献   

20.
Exposure of deaerated folic acid solutions containing an electron donor to UV radiation (310–390 nm, I = 0.4 W m−2) induced formation of dihydrofolic acid (DHFA), a photoexcitation which gave tetrahydrofolic acid (THFA). Only DHFA was formed in the presence of EDTA (Eo = +0.40 V), while the presence of stronger reductants—NADH (Eo = −0.32 V) and boron hydride (Eo = −0.48 V)—induced photoreduction to THFA. It was demonstrated that UV radiation had no effect on the THFA formylation, giving the coenzyme 5,10-methenyltetrahydrofolic acid and its transformation into another coenzyme, 5-formyltetrahydrofolic acid.  相似文献   

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