ABSTRACT
Amusurgus (Usgmona) excavatus Liu, Shi et Zhou, sp. nov. (China, Fujian) is described and illustrated with the male genitalia. Photos of habitus and ecological habitat are also included.
Subject(s)
Gryllidae/classification , Animal Distribution , Animal Structures/anatomy & histology , Animal Structures/growth & development , Animals , Body Size , China , Ecosystem , Female , Gryllidae/anatomy & histology , Gryllidae/growth & development , Male , Organ SizeABSTRACT
Thermal inactivation kinetics of Hypocrea orientalis ß-glucosidase and effect of glucose on thermostability of the enzyme have been determined in this paper. Kinetic studies showed that the thermal inactivation was irreversible and first-order reaction. The microscopic rate constants for inactivation of free enzyme and substrate-enzyme complex were both determined, which suggested that substrates can protect ß-glucosidase against thermal deactivation effectively. On the other hand, glucose was found to protect ß-glucosidase from heat inactivation to remain almost whole activity below 70°C at 20mM concentration, whereas the apparent inactivation rate of BG decreased to be 0.3×10(-3)s(-1) in the presence of 5mM glucose, smaller than that of sugar-free enzyme (1.91×10(-3)s(-1)). The intrinsic fluorescence spectra results showed that glucose also had stabilizing effect on the conformation of BG against thermal denaturation. Docking simulation depicted the interaction mode between glucose and active residues of the enzyme to produce stabilizing effect.
Subject(s)
Glucose/pharmacology , Hot Temperature , Hypocrea/enzymology , beta-Glucosidase/metabolism , Benzyl Alcohols/pharmacology , Cellobiose/pharmacology , Enzyme Stability/drug effects , Glucosides/pharmacology , Hydrolysis , Hypocrea/drug effects , Kinetics , Molecular Docking Simulation , Spectrometry, FluorescenceABSTRACT
Tyrosinase (EC 1.14.18.1) is the key enzyme of melanin synthesis and fruit-vegetable browning. The inhibition of benzylideneacetone, benzylacetone, and 4-phenyl-2-butanol on mushroom tyrosinase was first investigated. The results shown that these three compounds could effectively inhibit the enzyme activity sharply and the inhibitory effects were determined to be reversible. Their inhibitor concentrations leading to 50% activity lost values were determined to be 1.5, 2.8, and 1.1 mM for monophenolase and 2.0, 0.6, and 0.8 mM for diphenolase, respectively. For the monophenolase activity, all of these three compounds were mixed-type inhibitors, however, only 4-phenyl-2-butanol obviously lengthened the lag time. For the diphenolase activity, benzylideneacetone and benzylacetone were mixed-type inhibitors, while 4-phenyl-2-butanol was a noncompetitive type inhibitor. In conclusion, these compounds exhibited potent antityrosinase activities. This research would provide scientific evidence for the use of benzylideneacetone, benzylacetone, and 4-phenyl-2-butanol as antityrosinase agents.
Subject(s)
Acetone/analogs & derivatives , Agaricales/enzymology , Butanols/pharmacology , Butanones/pharmacology , Enzyme Inhibitors/pharmacology , Monophenol Monooxygenase/antagonists & inhibitors , Acetone/pharmacology , Food Additives/pharmacology , Kinetics , Levodopa/metabolism , Monophenol Monooxygenase/metabolism , Oxidation-Reduction/drug effects , Oxidoreductases/antagonists & inhibitors , Oxidoreductases/metabolismABSTRACT
In the present study the structure of proanthocyanidins from Polyalthia longifolia leaves was characterized with (13)C nuclear magnetic resonance, high performance liquid chromatography electrospray ionization mass spectrometry, and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analyses. The results showed that the proanthocyanidins were mixture of homopolymers of B-type procyanidins with degree of polymerization up to 14-mer. Furthermore, the antioxidant activity of the proanthocyanidins was studied through 2,2-diphenyl-1-picrylhydrazyl and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) free-radical scavenging activities, and ferric reducing/antioxidant power assays. In addition, antityrosinase activity of the proanthocyanidins was investigated. The IC50 for 2,2-diphenyl-1-picrylhydrazyl and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) free-radical scavenging activity of the proanthocyanidins were 89.32 ± 12.07 and 76.79 ± 5.88 µg/mL, respectively; the ferric reducing/antioxidant power value was 710.54 ± 142.82 mg ascorbic acid equivalent/g dry weight. The IC50 for antityrosinase activity was 773.09 ± 1.47 µg/mL. In conclusion, the proanthocyanidins from P. longifolia leaves exhibited potent antioxidant and antityrosinase activities. This research would provide scientific evidence for the use of proanthocyanidins from P. longifolia leaves as antioxidant and antityrosinase agents.
Subject(s)
Antioxidants/isolation & purification , Enzyme Inhibitors/isolation & purification , Monophenol Monooxygenase/antagonists & inhibitors , Plant Leaves/chemistry , Polyalthia/chemistry , Proanthocyanidins/isolation & purification , Proanthocyanidins/pharmacology , Antioxidants/chemistry , Ascorbic Acid/chemistry , Benzothiazoles/chemistry , Biflavonoids/chemistry , Biflavonoids/isolation & purification , Biphenyl Compounds/chemistry , Biphenyl Compounds/isolation & purification , Catechin/chemistry , Catechin/isolation & purification , Chromatography, High Pressure Liquid , Enzyme Inhibitors/chemistry , Enzyme Inhibitors/pharmacology , Free Radical Scavengers/chemistry , Inhibitory Concentration 50 , Magnetic Resonance Spectroscopy , Oxidation-Reduction , Picrates/chemistry , Picrates/isolation & purification , Proanthocyanidins/chemistry , Reducing Agents/chemistry , Spectrometry, Mass, Electrospray Ionization , Sulfonic Acids/chemistryABSTRACT
Proanthocyanidins were isolated from fruit stone of Chinese hawthorn (Crataegus pinnatifida Bge. var. major N.E.Br.). Their structures were analyzed and elucidated by methods of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and high performance liquid chromatography electrospray ionization mass spectrometry (HPLC-ESI-MS). The results demonstrated that these compounds are complicated mixtures of homo- and heteropolymers consisting of procyanidin/procyanidin gallate and prodelphinidin. They possessed structural heterogeneity in monomer units, polymer length, and interflavan linkage (A-type and B-type). Their antityrosinase and antioxidant activity were then investigated. The results revealed that they can inhibit tyrosinase activities, including the monophenolase activity and the diphenolase activity. In addition, proanthocyanidins possessed potent antioxidant activity. Our studies revealed that proanthocyanidins isolated from fruit stone of Chinese hawthorn may be applied in food, agriculture, pharmaceutical, and cosmetic industries.
Subject(s)
Antioxidants/chemistry , Crataegus/chemistry , Enzyme Inhibitors/chemistry , Monophenol Monooxygenase/antagonists & inhibitors , Plant Extracts/chemistry , Proanthocyanidins/chemistry , Antioxidants/isolation & purification , Enzyme Inhibitors/isolation & purification , Fruit/chemistry , Molecular Structure , Plant Extracts/isolation & purification , Proanthocyanidins/isolation & purificationABSTRACT
In this research, the conditions for extraction of phenolics from leaves of Ficus virens were optimized using response surface methodology (RSM). The extraction abilities of phenolics (EAP) and flavonoids (EAF), the 2,2-diphenyl-1-pierylhydrazyl (DPPH) free-radical scavenging potential, and the ferric reducing/antioxidant power (FRAP) were used as quality indicators. The results of single-factor experiments showed that temperature, ethanol concentration, extraction time, and the number of extraction cycles were the main influencing variables, and these provided key information for the central composite design. The results of RSM fitted well to a second degree polynomial model and more than 98% of the variability was explained. The ideal extraction conditions for EAP, EAF, DPPH free-radical scavenging potential, and FRAP were obtained. Considering the four quality indicators overall, the ideal extraction conditions were 58% ethanol at 57 °C for 37 min with three extraction cycles. At the ideal extraction conditions, the values of EAP, EAF, DPPH free-radical scavenging potential, and FRAP were 5.72%, 3.09%, 58.88 mg ascorbic acid equivalent (AAE)/g dry weight (DW), and 15.86 mg AAE/g DW, respectively. In addition, linear correlations were observed between EAP, EAF, and antioxidant potential.
Subject(s)
Ficus/chemistry , Phenols/chemistry , Phenols/isolation & purification , Plant Extracts/chemistry , Plant Leaves/chemistry , Antioxidants/chemistry , Biphenyl Compounds/chemistry , Chemistry Techniques, Analytical , Ethanol/chemistry , Flavonoids/chemistry , Free Radical Scavengers/chemistry , Models, Theoretical , Oxidative Stress , Picrates/chemistry , Solvents/chemistry , Temperature , Time FactorsABSTRACT
The inhibitory kinetics of furfuryl alcohol, furfural and furoic acid on mushroom tyrosinase have been investigated. The results showed that these furan compounds were reversible inhibitors of the enzyme. Furthermore, furfuryl alcohol and furfural were found to be mixed-type inhibitors while furoic acid is uncompetitive inhibitor. The inhibition constants have been confirmed and the order of the inhibiting ability was furfural>furoic acid>furfuryl alcohol. They indicate that the functional groups on the furan ring play a crucial role in the inhibition on the enzyme. In addition, it was also found that these furan compounds could inhibit the proliferation of Salmonella bacteria and Bacillus subtilis to different extents. The minimum inhibitory concentration (MIC) values of furfuryl alcohol, furfural and furoic acid against B. subtilis and S. bacteria were 0.115, 0.027, 0.015 and 0.115, 0.029, 0.009 µM, respectively. The minimum bactericidal concentration (MBC) values of that were 0.115, 0.027, 0.015 and 0.231, 0.121, 0.030 µM, respectively.
Subject(s)
Anti-Infective Agents/pharmacology , Bacillus subtilis/growth & development , Enzyme Inhibitors/pharmacology , Fungal Proteins/antagonists & inhibitors , Furaldehyde/pharmacology , Furans/pharmacology , Monophenol Monooxygenase/antagonists & inhibitors , Salmonella/growth & development , Anti-Infective Agents/chemistry , Enzyme Inhibitors/chemistry , Fungal Proteins/chemistry , Furaldehyde/chemistry , Furans/chemistry , Monophenol Monooxygenase/chemistryABSTRACT
NAGase (EC.3.2.1.52) from crustaceans has the important roles in immunity, molting and digestion of chitinous foods. In this paper, the effects of citric acid on the activity of NAGase from Litopenaeus vannamei for the hydrolysis of pNP-NAG have been studied. The results showed that appropriate concentrations of citric acid could lead to reversible inhibition on NAGase and IC(50) was estimated to be 5.00 +/- 0.35 mM. Using the plots of Lineweaver-Burk, the inhibition of NAGase by citric acid belongs to competitive type, the inhibitory equilibrium constant for citric acid binding with free NAGase, K(I), is 3.26 +/- 0.25 mM. The inhibitory kinetics of citric acid on NAGase in the appropriate concentrations of citric acid has been studied using the kinetic method of substrate reaction. The time course of NAGase for the hydrolysis of pNP-NAG in the presence of different concentrations of citric acid showed that at each citric acid concentration, the rate decreased with increasing time until a straight line was approached. The results show that the inhibition of NAGase by citric acid is a slow, reversible reaction with fractional remaining activity. The microscopic rate constants are determined for the reaction on citric acid with NAGase.
Subject(s)
Citric Acid/pharmacology , Enzyme Activation/drug effects , Enzyme Inhibitors/pharmacology , Penaeidae/enzymology , beta-N-Acetylhexosaminidases/metabolism , AnimalsABSTRACT
The effects of betaine on prawn beta-N-acetyl-D-glucosaminidase (NAGase) activity for the hydrolysis of p-nitrophenyl-N-acetyl- beta-D-glucosaminide (pNP-NAG) have been studied. The results showed that appropriate concentrations of betaine could lead to reversible inhibition against NAGase, and the IC(50) value was estimated to be 15.00 +/- 0.30 mM. The inhibitory kinetics assay showed that betaine was a mixed type inhibitor with a K(I) value of 9.17 +/- 0.85 mM and a K(IS) value of 45.58 +/- 2.52 mM. The inhibitory model was set, and the microscopic rate constants were determined using the kinetic method of the substrate reaction. The time course of the hydrolysis of pNP-NAG catalyzed by NAGase in the presence of different betaine concentrations showed that at each betaine concentration, the rate decreased with an increase in time until a straight line was approached, indicating that the inhibition of NAGase by betaine is a slow, reversible reaction with fractional residual activity. The fact that k(+0) is much larger than k(+0)(') indicated that the free enzyme molecule is more fragile than the enzyme-substrate complex against betaine. It is suggested that the presence of the substrate offers marked protection of NAGase against inhibition by betaine.
Subject(s)
Acetylglucosaminidase/antagonists & inhibitors , Acetylglucosaminidase/chemistry , Betaine/chemistry , Crustacea/enzymology , Enzyme Inhibitors/chemistry , Animals , Crustacea/chemistry , Kinetics , Protein BindingABSTRACT
Beta-N-acetyl-D-glucosaminidase (NAGase, EC.3.2.1.52), which catalyzes the cleavage of N-acetylglucosamine polymers, is a composition of chitinase and cooperates with endo-chitinase and exo-chitinase to disintegrate chitin into N-acetylglucosamine (NAG). In this investigation, A NAGase from green crab (Scylla serrata) was purified and the effects of dioxane on the enzyme activity for the hydrolysis of p-Nitrophenyl-N-acetyl-beta-D-glucosaminide (pNP-NAG) were studied. The results show that appropriate concentrations of dioxane can lead to reversible inactivation of the enzyme and the inactivation is classified as mixed type. The value of IC50, the dioxane (inactivator) concentration leading to 50% activity lost, is estimated to be 0.68%. The kinetics of inactivation of NAGase in the appropriate concentrations of dioxane solution has been studied using the kinetic method of the substrate reaction. The rate constants of inactivation have been determined. The results showed that k+0 is much larger than k'+0, indicating the free enzyme molecule is more fragile than the enzyme-substrate complex in the dioxane solution. It is suggested that the presence of the substrate offers marked protection of this enzyme against inactivation by dioxane.
Subject(s)
Acetylglucosaminidase/chemistry , Arthropod Proteins/chemistry , Brachyura/enzymology , Dioxanes/chemistry , Solvents/chemistry , Acetylgalactosamine/analogs & derivatives , Acetylgalactosamine/chemistry , Acetylglucosaminidase/antagonists & inhibitors , Animals , Arthropod Proteins/antagonists & inhibitors , Hydrolysis , Kinetics , SolutionsABSTRACT
RNAs of the leaves in Avicennia marina, which cultured in 50@1000 and 0@1000 salinity condition respectively, were isolated for mRNA differential display analysis. Screened by OligodT12 GC and eight 10-oligonucleotide arbitrary primers, differential cDNA fragments-csrg1(600 bp), csrg2(550 bp), csrg3(480 bp), only appeared in the leaves of Avicennia marina cultured in 50@1000 salinity condition. After detected by RNA dot hybridization, only csrg1 appeared the difference between the RNAs of the leaves in Avicennia marina cultured in 50@1000 and 0@1000 salinity condition respectively, and csrg1 was confirmed as the salt-tolerant cDNA. csrg1 was cloned and sequenced. After searching in Genbank, there were no similar sequences reported.
