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Lauric Acid (lauric + acid)
Selected AbstractsAntifungal activity of fatty acids and their monoglycerides against Fusarium spp. in a laboratory mediumINTERNATIONAL JOURNAL OF FOOD SCIENCE & TECHNOLOGY, Issue 2 2009Clelia Altieri Summary The antifungal activity of lauric, myristic and palmitic acids and their monoglycerides against fusaria was investigated. Data were modelled through a re-parameterized Gompertz equation and the Minimum Detection Time (MDT), i.e. the time (days) to attain 1 cm colony diameter, was evaluated. Lauric acid exerted a strong bioactivity against moulds; palmitic and myristic acids and their monoglycerides showed a moderate effectiveness and in a reversible manner. The results of this work could be considered quite promising; however, further investigations are proposed to validate these data in foods. [source] Disappearance of gender-related difference in the toxicity of benzotriazole ultraviolet absorber in juvenile ratsCONGENITAL ANOMALIES, Issue 4 2009Mutsuko Hirata-Koizumi ABSTRACT 2-(2,-hydroxy-3,,5,-di- tert -butylphenyl)benzotriazole (HDBB) is an ultraviolet absorber used in plastic resin products, such as building materials and automobile components. In oral repeated dose toxicity studies using 5- or 6-week-old rats, this chemical induced hepatic histopathological changes, such as hypertrophy accompanied with eosinophilic granular changes and focal necrosis of hepatocytes, and male rats showed nearly 25 times higher susceptibility to the toxic effects than females. Castration at approximately 4 weeks of age markedly reduced the sex-related variation in HDBB toxicity, but some difference, less than five times, remained between male and female castrated rats. Following oral HDBB administration to male and female juvenile rats from postnatal days 4,21, such gender-related difference in toxic susceptibility was not detected; therefore, it is speculated that the determinants of susceptibility to HDBB toxicity are differentiated between sexes after weaning. In young rats given HDBB, there was no gender-related difference in plasma HDBB concentration, and no metabolites were detected in the plasma of either sex. HDBB induced lauric acid 12-hydroxylase activity in the liver and this change was more pronounced in males than in females. These findings indicate that HDBB could show hepatic peroxisome proliferation activity, and the difference in the susceptibility of male and female rats to this effect might lead to marked gender-related differences in toxicity. [source] Integrated Microanalytical System Coupling Permeation Liquid Membrane and Voltammetry for Trace Metal Speciation.ELECTROANALYSIS, Issue 10 2004Optimization, Technical Description Abstract A new minicell coupling the liquid-liquid extraction technique called permeation liquid membrane (PLM) with an integrated Ir-based Hg-plated microelectrode array for voltammetric detection has been developed for the speciation of heavy metals in natural waters. Lead and cadmium have been used as model compounds. The PLM consists of a carrier (0.1,M 22DD+0.1,M lauric acid) dissolved in 1,:,1 mixture of toluene/phenylhexane held in the small pores (30,nm) of a hydrophobic polypropylene membrane (Celgard 2500). One side of this membrane is in contact with a flowing source solution, containing the metal ions of interest. An acceptor or strip solution (pyrophosphate) is placed on the other side of the PLM with the microelectrode array placed at 480,,m of the PLM. The analyte is transported by the carrier from the source solution to the strip solution. The originality of the new minicell is that accumulation in the strip solution is voltammetrically followed by the integrated microelectrode array in real time, and at low concentration level, using square-wave anodic stripping voltammetry (SWASV). In order to protect the Hg microelectrodes from the adsorption of the hydrophobic carrier, the microelectrodes are embedded in a thin gel layer (280,,m) of 1.5% LGL agarose gel containing 10% of hydrophobic silica particles C18. The choice of optimum conditions is discussed in details in this article. Due to the very small effective strip volume of the new cell (less than 1,,L), high enrichment factor can be obtained (e.g., 330 for Pb) after 2,hours of accumulation. No deaeration of the solutions is required for SWASV measurements. Detection limits under these conditions are 2,pM and 75,pM for Pb and Cd, respectively, using a voltammetric deposition time of 5,min. In addition, no fouling effects were observed with natural water samples. [source] Whey-derived free fatty acids suppress the germination of Candida albicans in vitroFEMS YEAST RESEARCH, Issue 2 2007Martin Clément Abstract Bovine whey from the cheese-making industry contains several bioactive factors that promote health and prevent disease. Although many efforts have been made over the years to show that immunoglobulins, lactoperoxidase, lactoferrin, lysosyme and small peptides present in whey have antimicrobial activities against several pathogenic microorganisms, such activities have not been investigated so far for the lipid fraction of whey. Here, we have used an in vitro assay-based fractionation procedure to show that free fatty acids derived from whey cream specifically inhibit the germination of Candida albicans, a morphologic change associated with pathogenicity. Further fractionation by HPLC demonstrated that this activity can be mainly attributed to lauric acid, myristoleic acid, linoleic acid and arachidonic acid. [source] Permeation of Sumatriptan Through Human Vaginal and Buccal MucosaHEADACHE, Issue 2 2000P. Van Der Bijl DSc Continued interest in the various routes by which sumatriptan may be administered prompted us to investigate its passage through buccal mucosa. Because human buccal mucosa is scarce, we proposed using the relatively abundant vaginal mucosa, which has been shown to have comparable diffusion rates for a number of widely varying molecules, as a model of buccal mucosa. In addition, by comparing these two tissues with respect to their permeability to sumatriptan, the human vaginal/buccal mucosa model could be further evaluated. Clinically healthy human vaginal and buccal mucosa specimens were used in the permeability studies. Permeability to sumatriptan was determined using a continuous flow-through diffusion system in the presence and absence of permeation enhancers. No statistically significant differences in permeability could be demonstrated for both mucosae toward sumatriptan. Flux values obtained in the absence and presence of glycodeoxycholate and lauric acid (1:1 molar ratio) to sumatriptan of buccal and vaginal mucosa, respectively, were not significantly different. The results obtained further support the hypothesis of the vaginal/buccal mucosal in vitro permeability model and suggest that this model may be used in conjunction with various absorption enhancers. Further studies on the buccal route of absorption of sumatriptan are thus warranted. [source] Heat transfer enhancement of fatty acids when used as PCMs in thermal energy storageINTERNATIONAL JOURNAL OF ENERGY RESEARCH, Issue 2 2008Muhsin Mazman Abstract Phase change materials (PCM) used in latent heat storage systems usually have very low thermal conductivities. This is a major drawback in maintaining the required heat exchange rate between PCM and heat transfer fluid. This paper investigates the enhancement of the heat transfer between PCM and heat transfer fluid, using high thermal conductivity as additives like stainless steel pieces, copper pieces and graphite,PCM composite material. In the experiments, palmitic,lauric acid (80:20) (PL) and stearic,myristic acid (80:20) (SM) were used as PCMs. Test results show that heat transfer enhancement of copper pieces was better at 0.05 Ls,1 flow rate compared to 0.025 Ls,1. Using copper as an additive increased the heat transfer rate 1.7 times for melting and 3.8 times for freezing when flow rate was 0.050 Ls,1. Decreasing the flow rate from 0.050 to 0.025 Ls,1, increased the melting times 1.3 times and freezing times 1.8 times, decreasing heat transfer rates accordingly. The best result of heat transfer enhancement was observed for the PCM,graphite composite. However, changing the flow rate did not affect the heat transfer rate when graphite was used as additive. Copyright © 2007 John Wiley & Sons, Ltd. [source] Cellular effects of monohydrochloride of l -arginine, N, -lauroyl ethylester (LAE) on exposure to Salmonella typhimurium and Staphylococcus aureusJOURNAL OF APPLIED MICROBIOLOGY, Issue 5 2004E. Rodríguez Abstract Aims:, Here we study the effect of monohydrochloride of l -arginine, N, -lauroyl ethylester (LAE), a cationic preservative derived from lauric acid and arginine, on the cell envelopes of Salmonella typhimurium and Staphylococcus aureus at sub-lethal concentration such as their respective minimal inhibitory concentrations, 32 and 8 ,g ml,1, respectively. Methods and Results:, Bacterial populations were studied by using transmission electron and fluorescence microscopy (TEM and FM), flow cytometry (FC) and ion-flux across the cellular membrane. Cell integrity was altered mainly in the outer membrane of S. typhimurium, but there was no significant change in the cytoplasm. However, in Staph. aureus, clear zones, abnormal septation and mesosome-like structures were observed in the cytoplasm. Bacterial populations were double-stained with propidium iodide (PI) and SYTO-13 for FC analysis. In S. typhimurium the proportion of damaged cells after 24 h was 97% and in Staph. aureus 56·3%. LAE induced transmembrane ion flux, the increase of potassium leakage after 30 min of contact was 7·7 and 3·34 ,g ml,1 for Staph. aureus and S. typhimurium, respectively. Membrane disruption was detected by measuring the proton flow across the membrane. Conclusions:, Disturbance in membrane potential and structural changes was caused by LAE, although cells were not disrupted. Significance and Impact of the Study:, This is the first time the cellular effects of LAE on bacterial cells were studied. [source] Separation of fatty acids from binary melts using physical vapour deposition (PVD)JOURNAL OF CHEMICAL TECHNOLOGY & BIOTECHNOLOGY, Issue 3 2009Young Han Kim Abstract BACKGROUND: The use of fatty acid mixtures, natural biochemical compounds, will be extended to various chemical industries for the production of a wide variety of products, and various mixtures of fatty acids are necessary for production. Separation of a binary fatty acid mixture of lauric acid and myristic acid using physical vapour deposition (PVD) on a cold quartz crystal resonator is examined. The extremely small amount of deposits can be measured with the quartz crystal resonator. The vapour phase is prepared by vaporizing a calculated composition of melt according to the vapour-liquid equilibrium (VLE). RESULTS: The composition of lauric acid in the melt and the melt temperature were utilized as operating variables in the PVD. The growth rate of deposit increases when melt temperature and the composition of lauric acid in the melt are increased. The composition of lauric acid in the deposit is significantly lower than that of the melt of 19% lauric acid, but the composition of lauric acid in the deposit is much higher than that of the melts of 50% and 75% lauric acid. CONCLUSION: The distribution coefficient of lauric acid between solid and vapour phases can be correlated as a function of the growth rate of deposit. The possibility of separation of fatty acid mixtures by PVD is suggested experimentally and theoretically. Copyright © 2008 Society of Chemical Industry [source] SHELF LIFE OF FRESH NOODLES AS AFFECTED BY THE FOOD GRADE MONOLAURIN MICROEMULSION SYSTEMJOURNAL OF FOOD PROCESS ENGINEERING, Issue 5 2008XIAOWEI FU ABSTRACT Food grade monolaurin (the glycerol monoester of lauric acid) microemulsions (MMEs) based on multicomponent mixtures and the effects of MMEs on the shelf life of fresh noodles were studied. A microemulsion system consisting of monolaurin (M), surfactants (Tween 20 and Tween 80, Wenzhou Qingming Chemical Engineering Company, Wenzhou, Zhejiang, China ) and cosurfactant (pentanol and dodecanol) dispersed in distilled water was used. The influence of surfactants on the phase behavior was studied in this article. Subsequently, two MMEs with large oil in water regions have demonstrated the enhanced antimicrobial activities against Bacillus subtilis, Escherichia coli, Aspergillus niger and Penicillium digitatum, compared to monolaurin (M) alone. In addition, the shelf life of fresh noodles prepared with monolaurin (M) alone was about 4 days, whereas MMEs extended shelf life by 10 days at least when stored at 37C. PRACTICAL APPLICATIONS The effect of monolaurin microemulsion (MME) as antimicrobial agents in foods, especially those neutral foods such as noodles, is obvious, and MME has demonstrated a much longer shelf life of fresh noodles than M alone. In addition, for those neutral foods other than fresh noodles, the application of MMEs can also be studied. [source] Resistance of Nutrient-Deprived Listeria monocytogenes 10403S and a ,sigB Mutant to Chemical Stresses in the Presence or Absence of OxygenJOURNAL OF FOOD SCIENCE, Issue 7 2008B. Lungu ABSTRACT:, Nutrient-deprived Listeria monocytogenes have increased resistance to processing control measures. Heat-stressed L. monocytogenes cells produce higher counts under anaerobic conditions and SigB reportedly contributes to the survival of environmentally stressed Gram-positive bacteria. In this study, a wild type (wt) strain, L. monocytogenes 10403S, and a ,sigB mutant, FSLA1-254, were stressed by starvation in phosphate buffered saline coupled with exposure to chemicals with/without oxygen. In the absence of chemicals, the mutant survived starvation almost as well as the wt, suggesting that the starvation survival response (SSR) in L. monocytogenes was SigB-independent. Conversely, in the presence of chemical stresses the SSR results differed depending on the chemical used. In the presence of sodium chloride (SC), both strains were able to express an SSR under aerobic conditions but not under anaerobic conditions. However, in the presence of sodium propionate (SP), the mutant yielded counts that were 2 log CFU/mL lower than the controls and their aerobic counterparts. In the presence of sodium lactate (SL), the mutant yielded counts that were approximately 3 log CFU/mL lower than the wt under anaerobic conditions. Thus, for the chemical stress produced by SC, the SSR appeared to be SigB-independent. The SSR of L. monocytogenes appeared to be SigB-dependent following exposure to SP or SL under anaerobic conditions. Following exposure to sodium diacetate or lauric acid, both strains were unable to express an SSR. No detectable CFUs were observed after 14 to 21 d under either aerobic or anaerobic incubation. Therefore, these 2 chemicals could be used in biocidal formulations against L. monocytogenes cells under aerobic or anaerobic conditions. [source] Phase equilibrium in supercritical CO2 mixtures using a modified Kwak-Mansoori mixing ruleAICHE JOURNAL, Issue 2 2004José O. Valderrama Abstract The mixing rules proposed by Kwak and Mansoori for the Peng-Robinson equation of state have been modified to describe vapor-liquid equilibrium in mixtures that contain supercritical CO2. Data in the literature for nine binary liquid,vapor systems containing supercritical CO2 are used for testing the modified models. The systems studied were binary mixtures containing carbon dioxide with lauric acid, palmitic acid, oleic acid, linoleic acid, 1-octanol, 1-decanol, 2-methyl-1-pentanol, ,-pinene, and limonene. The modifications studied included the introduction of a nonquadratic mixing rule proposed by one of the authors and a different interaction parameter for the volume constant of the equation of state. Compared to results in the literature, the Kwak-Mansoori combining rules and the proposed empirical modifications give lower deviations in correlating the solute concentration in the vapor phase, the most important variable for the design of supercritical extraction processes. Contrary to arguments found in the literature, it is also demonstrated that a severe test for an equation of state and its mixing rules is the correlation of the solute concentration in vapor,liquid mixtures. © 2004 American Institute of Chemical Engineers AIChE J, 50: 480,488, 2004 [source] Effects of Light and Dark Beer on Hepatic Cytochrome P-450 Expression in Male Rats Receiving Alcoholic Beverages as Part of Total Enteral NutritionALCOHOLISM, Issue 5 2005Mats Hidestrand Background: Alcoholic beverages contain many congeners in addition to ethanol. Therefore, consumption of alcoholic beverages may have considerably different effects on expression of hepatic microsomal monooxygenases than the relatively selective induction of cytochrome P-450 (CYP) 2E1 observed after consumption of pure ethanol. Methods: In the current study, we compared the effects of two beers: lager (a light roasted beer) and stout (a dark roasted beer) with those of an equivalent amount of pure ethanol on hepatic CYP expression. Beer or pure ethanol was part of a complete total enteral nutrition diet that was infused intragastrically into male Sprague Dawley rats for 21 days. At the end of the infusion period, rats were euthanized, and liver and intestinal microsomes were prepared. Expression and activity of CYP1A1/2, CYP2B1, CYP2E1, CYP3A, and CYP4A were assessed by Western immunoblotting and by using CYP enzyme,specific substrates, respectively. Results: mRNA and protein levels of CYP4A1 were elevated only in stout-treated animals. However, lauric acid 12-hydroxylase activity (a CYP4A-specific activity) was reduced (p, 0.05) in microsomes from lager- and stout-fed rats. After oxidation with potassium ferricyanide, this activity was significantly increased in microsomes from stout-fed animals. The relative expression of CYP2E1 and CYP2B1 and the activities toward p -nitrophenol, pentoxyresorufin, or benzyloxyresorufin did not differ between beers or compared with pure ethanol or controls. However, the mean expression of CYP1A2, CYP3A, and CYP4A apoproteins was greater in liver microsomes from stout-infused rats than in those from lager-infused rats, ethanol-infused rats, and diet controls (p, 0.05). In addition, although no significant differences were observed in ethoxyresorufin O-dealkylase (EROD), methoxyresorufin O-dealkylase (MROD), midazolam, or testosterone hydroxylase activities between groups, stout-infused rats had greater hepatic microsomal erythromycin N -demethylase activity than other groups (p, 0.05). Conclusions: Stout contains components other than ethanol that interact in a complex fashion with the monooxygenase system. [source] Optimisation of kojic acid monolaurate synthesis with lipase PS from Pseudomonas cepaciaJOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 6 2002Chee-Shan Chen Abstract To improve the instability of kojic acid in food and cosmetic use, the esterification of kojic acid catalysed by lipase from Pseudomonas cepacia (Amano PS) to synthesise kojic acid monolaurate (KAML) was investigated in this study. Response surface methodology (RSM) with a five-level/five-factor central composite rotatable design (CCRD) was employed to evaluate the effects of synthesis parameters such as reaction time (8,24,h), temperature (35 55,°C), enzyme amount (10,50%), substrate molar ratio of lauric acid to kojic acid (1:1,3:1) and added water content (0,20%) on the percentage molar conversion to KAML by direct esterification. Reaction time and added water content were the most important variables, while substrate molar ratio had less effect on percentage molar conversion. Based on canonical analysis and ridge maximum analysis, optimal synthesis conditions were reaction time 19,h, temperature 44,°C, enzyme amount 38%, substrate molar ratio 2:1 and added water content 10%. The predicted value was 85% and the actual experimental value 82% molar conversion. © 2002 Society of Chemical Industry [source] The identification of QTL that affect the fatty acid composition of milk on sheep chromosome 11ANIMAL GENETICS, Issue 3 2010M. García-Fernández Summary In this work, we analysed 11 genetic markers localized on OAR11 in a commercial population of Spanish Churra sheep to detect QTL that underlie milk fatty acid (FA) composition traits. Following a daughter design, we analysed 799 ewes distributed in 15 half-sib families. Eight microsatellite markers and three novel SNPs identified in two genes related to fatty acid metabolism, acetyl-CoA carboxylase , (ACACA) and fatty acid synthase (FASN), were genotyped in the whole population under study. The phenotypic traits considered in the study included 22 measurements related to the FA composition of the milk and three other milk production traits (milk protein percentage, milk fat percentage and milk yield). Across-family regression analysis revealed four significant QTL at the 5% chromosome-wise level influencing contents of capric acid (C10:0), lauric acid (C12:0), linoleic conjugated acid (CLA) and polyunsaturated fatty acids (PUFA) respectively. The peaks of the QTL affecting C10:0 and PUFA contents in milk map close to the FASN gene, which has been evaluated as a putative positional candidate for these QTL. The QTL influencing C12:0 content reaches its maximum significance at 58 cM, close to the gene coding for the glucose-dependent insulinotropic polypeptide. We were not able to find any candidate genes related to fat metabolism at the QTL influencing CLA content, which is located at the proximal end of the chromosome. Further research efforts will be needed to confirm and refine the QTL locations reported here. [source] Stearoyl-CoA desaturase 1 genotype and stage of lactation influences milk fatty acid composition of Canadian Holstein cowsANIMAL GENETICS, Issue 5 2009P. M. Kgwatalala Summary Single nucleotide polymorphisms in the coding region of the bovine stearoyl-CoA desaturase 1 gene have been predicted to result in p.293A (alanine at amino acid 293) and p.293V (valine at amino acid 293) alleles at the stearoyl-CoA desaturase1 locus. The objectives of this study were to evaluate the extent to which genotypes at the stearoyl-CoA desaturase 1 locus and stage of lactation influence milk fatty acid composition in Canadian Holstein cows. Cows with the p.293AA genotype had higher C10 index, C12 index and C14 index and higher concentrations of C10:1 (10 carbon fatty acid with one double bond), C12:1 (12 carbon fatty acid with one double bond) and myristoleic acid (C14:1) compared with the p.293AV or p.293VV cows. Cows had higher C18 index and total index, and lower C10 index, C12 index, C14 index and CLA index during early lactation compared with the subsequent lactation stages. Early lactation was also characterized by higher concentrations of oleic acid (C18:1 cis -9), vaccenic acid (C18:1 trans -11), linoleic acid (C18:2), monounsaturated fatty acids and total polyunsaturated fatty acids, and lower concentrations of capric acid (C10:0), C10:1, lauric acid (C12:0), C12:1, myristic acid (C14:0), myristoleic acid (C14:1), palmitic acid (C16:0) and total saturated fatty acids compared with the subsequent lactation stages. Neither the stearoyl-CoA desaturase 1 genotype nor the stage of lactation had an influence on conjugated linoleic acid concentrations in milk. [source] Influences of dietary fatty acid profile on growth, body composition and blood chemistry in juvenile fat cod (Hexagrammos otakii Jordan et Starks)AQUACULTURE NUTRITION, Issue 1 2009S.-M. LEE Abstract This study was conducted to investigate the influence of dietary lipid source and n-3 highly unsaturated fatty acids (n-3 HUFA) level on growth, body composition and blood chemistry of juvenile fat cod. Triplicate groups of fish (13.2 ± 0.54 g) were fed the diets containing different n-3 HUFA levels (0,30 g kg,1) adjusted by either lauric acid or different proportions of corn oil, linseed oil and squid liver oil at 100 g kg,1 of total lipid level. Survival was not affected by dietary fatty acids composition. Weight gain, feed efficiency and protein efficiency ratio (PER) of fish fed the diets containing squid liver oil were significantly (P < 0.05) higher than those fed the diets containing lauric acid, corn oil or linseed oil as the sole lipid source. Weight gain, feed efficiency and PER of fish increased with increasing dietary n-3 HUFA level up to 12,16 g kg,1, but the values decreased in fish fed the diet containing 30 g kg,1 n-3 HUFA. The result of second-order polynomial regression showed that the maximum weight gain and feed efficiency could be attained at 17 g kg,1 n-3 HUFA. Plasma protein, glucose and cholesterol contents were not affected by dietary fatty acids composition. However, plasma triglyceride content in fish fed the diet containing lauric acid as the sole lipid source was significantly (P < 0.05) lower than that of fish fed the other diets. Lipid content of fish fed the diets containing each of lauric acid or corn oil was lower than that of fish fed the diets containing linseed oil or squid liver oil only. Fatty acid composition of polar and neutral lipid fractions in the whole body of fat cod fed the diets containing various levels of n-3 HUFA were reflected by dietary fatty acids compositions. The contents of n-3 HUFA in polar and neutral lipids of fish increased with an increase in dietary n-3 HUFA level. These results indicate that dietary n-3 HUFA are essential and the diet containing 12,17 g kg,1 n-3 HUFA is optimal for growth and efficient feed utilization of juvenile fat cod, however, excessive n-3 HUFA supplement may impair the growth of fish. [source] Interaction of dexloxiglumide, a cholecystokinin type-1 receptor antagonist, with human cytochromes P450BIOPHARMACEUTICS AND DRUG DISPOSITION, Issue 4 2004Michael Hall Abstract Dexloxiglumide (DEX) is a cholecystokinin type-1 receptor antagonist under development for the treatment of constipation-predominant irritable bowel syndrome. Studies of the potential interaction of DEX with human cytochromes P450 (CYPs) were conducted in vitro. DEX (300µM), both with and without a 15-min pre-incubation, was incubated with pooled human liver microsomes and substrates selective for each of eight CYPs. This resulted in >30% inhibition of tolbutamide 4-methyl-hydroxylase (CYP2C9/10) and lauric acid 11-hydroxylase (CYP2E1) activities. Mean Ki (SD) for CYP2C9/10 and CYP2E1 were 69.0 (24.3) and 426 (60)µM, respectively. Incubations of [14C]DEX with pooled human liver microsomes produced one major phase I metabolic fraction, with Vmax=131 pmol/min/mg protein and Km=23.7µM. Further incubations with (i) liver microsomes from 16 individual donors (correlation analysis), (ii) SupersomesÔ and (iii) selective chemical inhibitors, implicated CYP3A4/5, CYP2B6 and CYP2C9 in the formation of this component. Thus, DEX interacts with CYP2C9 both as inhibitor (Ki=69.0µM) and as substrate in vitro. However, based on the maximum concentration (27µM) after repeated oral doses of 200mg t.i.d. and the unbound fraction (0.03) of DEX in human plasma, no clinically relevant metabolic interactions with other CYP substrates are predicted. Copyright © 2004 John Wiley & Sons, Ltd. [source] Peroxidase activity of bacterial cytochrome P450 enzymes: Modulation by fatty acids and organic solventsBIOTECHNOLOGY JOURNAL, Issue 8 2010Kersten S. Rabe Abstract The modulation of peroxidase activity by fatty acid additives and organic cosolvents was determined and compared for four bacterial cytochrome P450 enzymes, thermostable P450 CYP119A1, the P450 domain of CYP102A1 (BMP), CYP152A1 (P450bs,), and CYP101A1 (P450cam). Utilizing a high-throughput microplate assay, we were able to readily screen more than 100 combinations of enzymes, additives and cosolvents in a convenient and highly reproducible assay format. We found that, in general, CYP119A1 and BMP showed an increase in peroxidative activity in the presence of fatty acids, whereas CYP152A1 revealed a decrease in activity and CYP101A1 was only slightly affected. In particular, we observed that the conversion of the fluorogenic peroxidase substrate Amplex Red by CYP119A1 and BMP was increased by a factor of 38 or 11, respectively, when isopropanol and lauric acid were present in the reaction mixture. The activity of CYP119A1 could thus be modulated to reach more than 90% of the activity of CYP152A1 without effectors, which is the system with the highest peroxidative activity. For all P450s investigated we found distinctive reactivity patterns, which suggest similarities in the binding site of CYP119A1 and BMP in contrast with the other two proteins studied. Therefore, this study points towards a role of fatty acids as activators for CYP enzymes in addition to being mere substrates. In general, our detailed description of fatty acid- and organic solvent-effects is of practical interest because it illustrates that optimization of modulators and cosolvents can lead to significantly increased yields in biocatalysis. [source] |