Acid Oxidation System Research Articles

Antioxidant properties of papain hydrolysates of wheat gluten in different oxidation systems

Enzymatic hydrolysis was used for preparing hydrolysates from wheat gluten which is by-product during production of wheat starch. The enzyme used for the hydrolysis was papain. The hydrolysate was separated based on the molecular weight of the peptides by membrane ultrafiltration (UF) with a molecular weight cut-off of 5 kDa into permeate (P) and retentate (5-K) fractions. The antioxidative activities of the hydrolysate and its UF fractions were investigated by using the TBA method and scavenging effect of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical. The three fractions showed strong antioxidative activities in the linoleic acid oxidation system, and exhibited DPPH radical scavenging activity. The antioxidative activity of the P fraction was almost the same as that of vitamin E at pH 7.0. The molecular weight distribution of the P fraction was concentrated in 4.2 kDa (86.5%) after gel permeation chromatography fractionation using an HPLC system.The P and 5-K fractions had higher surface hydrophobicities ( H 0) at pH7.0 compared with the hydrolysate. The resulting UF fractions were superior to the hydrolysate in terms of antioxidative activities.

Transcription Coactivator PRIP, the Peroxisome Proliferator-Activated Receptor (PPAR)-Interacting Protein, Is Redundant for the Function of Nuclear Receptors PPARα and CAR, the Constitutive Androstane Receptor, in Mouse Liver

Disruption of the genes encoding for the transcription coactivators, peroxisome proliferator-activated receptor (PPAR)-interacting protein (PRIP/ASC-2/RAP250/TRBP/NRC) and PPAR-binding protein (PBP/TRAP220/DRIP205/MED1), results in embryonic lethality by affecting placental and multiorgan development. Targeted deletion of coactivator PBP gene in liver parenchymal cells (PBP(LIV-/-)) results in the near abrogation of the induction of PPARalpha and CAR (constitutive androstane receptor)-regulated genes in liver. Here, we show that targeted deletion of coactivator PRIP gene in liver (PRIP(LIV-/-)) does not affect the induction of PPARalpha-regulated pleiotropic responses, including hepatomegaly, hepatic peroxisome proliferation, and induction of mRNAs of genes involved in fatty acid oxidation system, indicating that PRIP is not essential for PPARalpha-mediated transcriptional activity. We also provide additional data to show that liver-specific deletion of PRIP gene does not interfere with the induction of genes regulated by nuclear receptor CAR. Furthermore, disruption of PRIP gene in liver did not alter zoxazolamine-induced paralysis, and acetaminophen-induced hepatotoxicity. Studies with adenovirally driven EGFP-CAR expression in liver demonstrated that, unlike PBP, the absence of PRIP does not prevent phenobarbital-mediated nuclear translocation/retention of the receptor CAR in liver in vivo and cultured hepatocytes in vitro. These results show that PRIP deficiency in liver does not interfere with the function of nuclear receptors PPARalpha and CAR. The dependence of PPARalpha- and CAR-regulated gene transcription on coactivator PBP but not on PRIP attests to the existence of coactivator selectivity in nuclear receptor function.

L-Carnitine is Synthesized in the Human Fetal–Placental Unit: Potential Roles in Placental and Fetal Metabolism

Carnitine plays an indispensable role in fatty acid oxidation. Previous studies revealed that fetal carnitine is derived from the mother via transplacental transfer. Recent studies demonstrated the presence and importance of an active fatty acid oxidation system in the human placenta and in the human fetus. In view of these findings we decided to study carnitine metabolism in the fetal-placental unit by measuring carnitine metabolites, intermediary metabolites of carnitine biosynthesis, as well as the activity of carnitine biosynthesis enzymes in human term placenta, cord blood and selected embryonic and fetal tissues (5-20 weeks of development). Placenta contained low but detectable activity of gamma-butyrobetaine dioxygenase. This enzyme, which was considered to be expressed only in kidney, liver and brain, catalyzes the last step in the carnitine biosynthesis pathway. In addition, our results show that human fetal kidney, liver and spinal cord already have the capacity to synthesize carnitine. The ability of the placenta and fetus to synthesize carnitine suggests that in circumstances when maternal carnitine supply is limited, carnitine biosynthesis by the fetal-placental unit may supply sufficient carnitine for placental and fetal metabolism.

The Structure of Energy Supply for Tissue Adaptation in Small Mammals Differing in Ecological Specialization

Multivariate factor analysis of the data on liver bioenergetics of mountain and forest voles has been performed. The specific features of tissue energy metabolism are best differentiated in the space of characters pertaining to the succinic acid oxidation system. The role of the dominant oxidation substrate in the structure of energy supply for the adaptation of tissue to a set of environmental factors in specialized and widespread species is discussed.

Hepatic Peroxisomal Fatty Acid β-Oxidation Is Regulated by Liver X Receptor α

Peroxisomes are the exclusive site for the beta-oxidation of very-long-chain fatty acids of more than 20 carbons in length (VLCFAs). Although the bulk of dietary long-chain fatty acids are oxidized in the mitochondria, VLCFAs cannot be catabolized in mitochondria and must be shortened first by peroxisomal beta-oxidation. The regulation of peroxisomal, mitochondrial, and microsomal fatty acid oxidation systems in liver is mediated principally by peroxisome proliferator-activated receptor alpha (PPARalpha). In this study we provide evidence that the liver X receptor (LXR) regulates the expression of the genetic program for peroxisomal beta-oxidation in liver. The genes encoding the three enzymes of the classic peroxisomal beta-oxidation cycle, acyl-coenzyme A (acyl-CoA) oxidase, enoyl-CoA hydratase/L-3-hydroxyacyl-CoA dehydrogenase, and 3-ketoacyl-CoA thiolase, are activated by the LXR ligand, T0901317. Accordingly, administration of T0901317 in mice promoted a dose-dependent and greater than 2-fold increase in the rate of peroxisomal beta-oxidation in the liver. The LXR effect is independent of PPARalpha, because T0901317-induced peroxisomal beta-oxidation in the liver of PPARalpha-null mice. Interestingly, T0901317-induced peroxisomal beta-oxidation is dependent on the LXRalpha isoform, but not the LXRbeta isoform. We propose that induction of peroxisomal beta-oxidation by LXR agonists may serve as a counterregulatory mechanism for responding to the hypertriglyceridemia and liver steatosis that is promoted by potent LXR agonists in vivo; however, additional studies are warranted.

An Improved Desulfurization Process Based on H2O2/Formic Acid Oxidation System Followed by Liquid-Liquid Extraction. Part 1. Coker Gas Oil Feedstocks

An improved desulfurization process for CGO (coker gas oil) has been investigated on the basis of the oxidation of sulfur compounds by using hydrogen peroxide and formic acid as oxidation reagents. Thiophenic sulfur in CGO was determined via that thioethers could be oxidized to sulfones with KIO3 in acidic medium and LC (liquid chromatography) method. Aiming at reducing the wasteful decomposition of hydrogen peroxide in the selective oxidation desulfurization H2O2/formic acid system, some additives used in the paper or pulp industry, including EDTA, sodium silicate, and magnesium silicate, were used to improve the oxidative efficiency. The following extraction results of oxidized CGO used DMF, furfural, and NMP as solvent showed that EDTA could stabilize desulfurization liquors by complexing transition metals that catalyze hydrogen peroxide decomposition, thus making oxidation and extraction processes more effective.

Antioxidative ability in a linoleic acid oxidation system and scavenging abilities against active oxygen species of enzymatic hydrolysates from pollen Cistus ladaniferus

The enzymatic hydrolysates from pollen Cistus ladaniferus were digested and prepared using three kinds of enzymes (pepsin, trypsin, and papain) and the antioxidative properties were investigated. The yields, total phenolic contents, and protein contents of these hydrolysates were as follows: yields (about 21-45%), total phenolics (10.39-14.33 microg/mg sample powder), and proteins (129.62-137.35 microg/mg sample powder), respectively. The hydrolysates possessed strongly antioxidative and scavenging abilities against reactive oxygen species. The present studies revealed that hydrolysates from honeybee-collected pollen are of benefit not only to the materials of health food diets, but also to patients with various diseases such as cancer, cardiovascular diseases, and diabetes.

Alteration of the Malonyl-CoA/Carnitine Palmitoyltransferase I Interaction in the β-Cell Impairs Glucose-Induced Insulin Secretion

Carnitine palmitoyltransferase I, which is expressed in the pancreas as the liver isoform (LCPTI), catalyzes the rate-limiting step in the transport of fatty acids into the mitochondria for their oxidation. Malonyl-CoA derived from glucose metabolism regulates fatty acid oxidation by inhibiting LCPTI. To examine directly whether the availability of long-chain fatty acyl-CoA (LC-CoA) affects the regulation of insulin secretion in the beta-cell and whether malonyl-CoA may act as a metabolic coupling factor in the beta-cell, we infected INS(832/13) cells and rat islets with an adenovirus encoding a mutant form of LCPTI (Ad-LCPTI M593S) that is insensitive to malonyl-CoA. In Ad-LCPTI M593S-infected INS(832/13) cells, LCPTI activity increased sixfold. This was associated with enhanced fatty acid oxidation, at any glucose concentration, and a 60% suppression of glucose-stimulated insulin secretion (GSIS). In isolated rat islets in which LCPTI M593S was overexpressed, GSIS decreased 40%. The impairment of GSIS in Ad-LCPTI M593S-infected INS(832/13) cells was not recovered when cells were incubated with 0.25 mmol/l palmitate, indicating the deep metabolic influence of a nonregulated fatty acid oxidation system. At high glucose concentration, overexpression of a malonyl-CoA-insensitive form of LCPTI reduced partitioning of exogenous palmitate into lipid esterification products and decreased protein kinase C activation. Moreover, LCPTI M593S expression impaired K(ATP) channel-independent GSIS in INS(832/13) cells. The LCPTI M593S mutant caused more pronounced alterations in GSIS and lipid partitioning (fat oxidation, esterification, and the level of nonesterified palmitate) than LCPTI wt in INS(832/13) cells that were transduced with these constructs. The results provide direct support for the hypothesis that the malonyl-CoA/CPTI interaction is a component of a metabolic signaling network that controls insulin secretion.

Identification of promethin and PGLP as two novel up-regulated genes in PPARγ1-induced adipogenic mouse liver

Peroxisome proliferator-activated receptor (PPAR) isoforms, α, γ and β/δ, function as important lipid sensors as well as key regulators of energy homeostasis. PPARα plays a dynamic role in energy combustion by transcriptionally upregulating fatty acid oxidation systems primarily in liver, whereas PPARγ functions as a regulator of adipogenesis and lipid storage. Overexpression of PPARγ, using adenoviral expression approach, in PPARα deficient mouse liver results in hepatic steatosis with concurrent expression of adipocyte specific genes. In this study, to gain a global molecular understanding of PPARγ1-induced gene expression in liver, we have analyzed gene expression profiles using the Affymetrix GeneChip mouse expression array set 430, that enables a comprehensive gene expression profiling with >39,000 transcripts. Microarray data analysis provided us with over 278 genes up-regulated fourfold or higher, and 121 genes down-regulated fourfold or higher in liver with PPARγ-induced hepatic adiposis. We have found 101 uncharacterized genes out of 278 up-regulated and 29 uncharacterized among the down-regulated gene categories, respectively. Of 177 functionally characterized candidate genes in the up-regulated category many appear to be involved in adipogenesis, lipid metabolism and signal transduction. To focus attention on the uncharacterized genes in the up-regulated category, we cloned the full-length cDNAs of two novel candidates, which we designated as promethin and PGLP. Promethin, a 15-kDa cytosolic protein, is not normally expressed in liver but induced robustly in liver with hepatic adiposis caused by PPARγ overexpression. PGLP, which encodes a 38 kDa cytoplasmic membranous protein, is a low abundant transcript in normal liver, but induced dramatically following PPARγ overexpression. The expression of these two genes was not increased in fatty livers induced by fasting or choline deficiency. The identification of these and other novel PPARγ-target genes should provide a basis for understanding the molecular mechanisms underlying energy storage and lipid homeostasis.

Occurrence of Glutathione Sulphydryl (GSH) and Antioxidant Activities in Probiotic Lactobacillus spp.

The antioxidative ability on the basis of reduced glutathione sulphydryl level, the inhibition activities of linoleic acid peroxidation of cell free extract of Lactobacillus spp. and the effects of types of media and growth phase of the cells on the cellular GSH level have been determined. Correlation between reduced glutathione sulphydryl level and antioxidative ability of Lactobacillus spp. was analyzed: Lactobacillus casei HY 2782 contained 25.15 µmole/g of GSH, the cellular GSH level of L. casei HY 2782 reached maximum after 24 h of cultivation and tended to decrease on further cultivation up to 72 h. There was a significantly higher level of cellular GSH when grown in de Man Rogosa and Sharpe (MRS) broth than in tryptone phytone yeast extract (TPY) broth or bromcresol pruple dextrose (BCP) broth (p<0.05). The antioxidant activity of cell free extract of Lactobacillus spp. have been shown to be significantly different among strains in the inhibition of linoleic acid peroxidation by thiobarbituric acid (TBA) test (p<0.01). L. casei HY 2782 and L. acidophilus ATCC 4356 revealed a high degree of antioxidative effect in linoleic acid oxidation system. Spearmans' rank correlation coefficient between inhibitory activity on linoleic acid peroxidation and cellular GSH levels of Lactobacillus spp. was 0.65, which means a significant positive correlation. (Asian-Aust. J. Anim. Sci. 2004. Vol 17, No. 11 : 1582-1585)

Antioxidant activity of egg-yolk protein hydrolysates in a linoleic acid oxidation system

The antioxidant activities of egg-yolk protein hydrolysates in a linoleic acid system were investigated. Egg-yolk protein hydrolysates were prepared by enzymic hydrolysis of fat-free egg-yolk protein, which led to the main peak of the molecular mass distribution of lower than 1000. Egg-yolk protein hydrolysates showed strong antioxidant activity in a linoleic acid oxidation system as compared with the egg-yolk protein or amino acids mixture in which egg-yolk protein hydrolysates were constituted. The effect of the sample was concentration-dependent. Egg-yolk protein hydrolysates also showed strong antioxidant activities on cookies containing linoleic acid. These results suggest that egg-yolk protein hydrolysates could be a suitable natural antioxidant for preventing the oxidation of polyunsaturated fatty acids and related food ingredients.

Free radical scavenging and antioxidative activity of caffeic acid amide and ester analogues: structure-activity relationship.

The structure-activity relationships of synthetic caffeic acid amide and ester analogues as potential antioxidants and free radical scavengers have been investigated. The 2,2-diphenyl-1-picrylhydrazyl radical (DPPH.) scavenging activity of the test compounds was N-trans-caffeoyl-L-cysteine methyl ester (5) > N-trans-caffeoyldopamine (4) > N-trans-caffeoyltyramine (3) > N-trans-caffeoyl-beta-phenethylamine (2) > Trolox C (8) > caffeic acid phenethyl ester (1) > caffeic acid (6) > ferulic acid (7). This established that the radical scavenging activity of the compounds increased with increasing numbers of hydroxyl groups or catechol moieties and also with the presence of other hydrogen-donating groups (-NH, -SH). The antioxidative activity of the compounds was also investigated in an emulsified linoleic acid oxidation system accelerated by 2,2'-azobis(2-amidinopropane) dihydrochloride. The order was 1 > 2 > 4 > 3 > or = 5 > 6 > 8 > 7. Therefore, in the emulsion system, the antioxidative activity of the test compounds depends not only on the hydroxyl groups or catechol rings but also on the partition coefficient (log P) or hydrophobicity of the compounds. This supports the concept that hydrophobic antioxidants tend to exhibit better antioxidative activity in an emulsion system.

Relevance of fatty acid oxidation in regulation of the outer mitochondrial membrane permeability for ADP

The present study on saponin-treated rat heart muscle fibers has revealed a new function of the fatty acid oxidation system in the regulation of the outer mitochondrial membrane (OMM) permeability for ADP. It is found that oxidation of palmitoyl-CoA+carnitine, palmitoyl-L-carnitine and octanoyl-L-carnitine (alone or in combination with pyruvate+malate) dramatically decreased a very high value of apparent K(m) of oxidative phosphorylation for ADP. Octanoyl-D-carnitine, as well as palmitate, palmitoyl-CoA, and palmitoyl-L-carnitine were not effective in this respect, when their oxidation was prevented by the absence of necessary cofactors or blocked with rotenone. Our data suggest that oxidation, but not transport of fatty acids into mitochondria, induces an increase in the OMM permeability for ADP.

A Trail of Research from Lipoic Acid to α-Keto Acid Dehydrogenase Complexes

A Trail of Research from Lipoic Acid to α-Keto Acid Dehydrogenase Complexes

Factors Influencing Fetal Growth

After completing this article, readers should be able to: 1. Describe the roles of insulin-like growth factor on fetal growth and development. 2. Delineate components of “maternal constraint.” 3. Explain the role of the placenta in regulating fetal growth. 4. Describe the effects of fetal glucose, amino acid, and fat supply on fetal growth. The principal determinants of fetal growth are fetal genotype and in utero environment. Environmental factors include maternal and paternal genetics, maternal size, and the capacity of the placenta to provide nutrients to the fetus. These environmental factors interact with the intrinsic growth pattern of the fetus, yielding a particular rate and composition of fetal growth. Most of the variation in fetal growth in a population is due to variations in environmental factors, not the fetal genome, although a genetically abnormal fetus clearly might not grow as well as a normal fetus if affected genes include those that are important for growth. Many genes contribute to fetal growth and birthweight. Techniques in which specific genes can either be deleted (“knockouts”) or overexpressed have led to greater understanding of how some of these genes regulate fetal growth. Such studies have shown that both maternal and paternal influences are present during fetal development and are passed on to the developing fetus by spermatozoa or oogonia by a mechanism called imprinting. Although the maternal genetic composition exerts greater influence than fetal genotype in the overall regulation of fetal growth, both maternal and paternal genomes are important in fetal growth and development (1)(2). For example, gynogenetic zygotes (two maternal genome copies) lead to underdeveloped extraembryonic tissues but well-developed embryos. The more modest regulation offered by the paternal genotype is essential for trophoblast development. For example, zygote nuclear transfer experiments have shown that androgenetic zygotes (two paternal genome copies) develop extensive trophoblast tissues but …

Effects of organophosphorous compounds on fatty acid compositions and oxidative phosphorylation system in the brain of rats

Effects of organophosphorous compounds, O,O-dimethyl O-4-nitro-m-tolyl phosphorothioate (MEP), O-ethyl O-p-nitrophenyl phenylphosphonothioate (EPN) and O-(4-cyanophenyl) O-ethyl phenylphosphonothioate (CYP), on the fatty acid composition and the subsequent effects on the oxidative phosphorylation system in the brain of rats were studied. After 6 days exposure in pesticides, polyunsaturated fatty acids in brain free fatty acid fractions of CYP treated rats decreased, and the unsaturation index in them were lower than those in the control rats. The polyunsaturated/saturated ratio (P/S ratio) in brain total lipids of CYP treated rats was lower than that in the control. The fatty acid composition in the brain of EPN treated rats had the same inclination as that of CYP treated rats. In the P/S ratio and unsaturation index in serum no difference was observed between CYP treated rats and the control, therefore CYP could affect the fatty acid composition in the rat brain directly. Free fatty acid contents in the brain of EPN and CYP treated rats decreased after 6 days exposure. Activities of complex I of brains were significantly higher in the EPN and CYP exposed rats than in the control rats in spite of the fact that no difference of ATP productivity was observed between them. These results suggest that EPN and CYP may affect the free fatty acid content, especially the polyunsaturated fatty acid content and consequently the enzyme activities in the oxidative phosphorylation system in the brain. Those phenomena, however, were not observed after 28 days exposure in pesticide, therefore those effects may be a passing phenomenon in an acute stage.

A two-step bioconversion process for vanillin production from ferulic acid combining Aspergillus niger and Pycnoporus cinnabarinus

A two-step bioconversion process of ferulic acid to vanillin was elaborated combining two filamentous fungi, Aspergillus niger and Pycnoporus cinnabarinus. In the first step, A. niger transformed ferulic acid to vanillic acid and in the second step vanillic acid was reduced to vanillin by P. cinnabarinus. Ferulic acid metabolism by A. niger occurred essentially via the propenoic chain degradation to lead to vanillic acid, which was subsequently decarboxylated to methoxyhydroquinone. In 3-day-old cultures of P. cinnabarinus supplied with vanillic-acid-enriched culture medium from A. niger as precursor source, vanillin was successfully produced. In order to improve the yields of the process, sequential additions of precursors were performed. Vanillic acid production by A. niger from ferulic acid reached 920 mg l −1 with a molar yield of 88% and vanillin production by P. cinnabarinus from vanillic acid attained 237 mg l −1 with a molar yield of 22%. However, the vanillic acid oxidative system producing methoxyhydroquinone was predominant in P. cinnabarinus cultures, which explained the relatively low level in vanillin.

Biomimetic oxidation of plant protecting agents.

The suitability of two in vitro oxidation systems as chemical models for the biological degradation of plant protecting agents has been investigated. As representative herbicides diclofop, fenoxaprop, isoproturon, linuron and monolinuron have been oxidised by two systems, the Fentons' reagent and the ascorbic acid oxidation system (AAOS) and the results compared to those of the known metabolic pathways of these compounds. The herbicides have been oxidised by Fentons' reagent (hydroxy radicals). The main products were isolated by preparative scale HPLC and identified with (1)H-NMR and MS. Some of the products have been identified by comparing their retention times and UV/Vis-spectra to those of standard compounds. Several products known from biological degradation are also found after chemical oxidation, however, notable differences between the two pathways have been observed, for instance in the case of diclofop. Oxidation by the AAOS leads to comparable results. Reaction rates for the oxidation with the AAOS have been studied and compared with data known from degradation studies of the herbicides in soil. Compounds which are slowly degraded in soil are oxidised more slowly in the biomimetic process than those with a fast degradation in soil.

Reducing activity level of alliin.

Alliin has been reported to have antioxidative activity. We tried to reexamine the antioxidative activity of alliin with a purified alliin preparation, and found that it has no antioxidative activity in a linoleic acid oxidation system. The reducing activity of alliin tested by the ECD method was also weak in showing antioxidative activity.

Nitric oxide regulates peroxisomal enzyme activities.

We have previously shown that peroxisomes are involved in the production and detoxification of reactive oxygen species and that peroxisomal functions are damaged by such oxygen species. Since nitric oxide is not only a cellular messenger, but also a free radical, it would be interesting to detect a connection between nitric oxide levels and peroxisomal enzyme activities. To determine if nitric oxide has an effect on the activities of peroxisomal functions and whether this effect is based solely on its chemical properties as reactive oxygen species or its action as a second messenger, effectors of the cellular nitric oxide level were applied to a cell model (human skin fibroblasts in culture) or directly to the enzymatic assays or both. If applied to the monolayer at non-cytotoxic concentrations, N-nitro-L-arginine methyl ester hydrochloride, an inhibitor of nitric oxide synthase (EC 1.14.13.39), increased catalase (EC 1.11.1.6) activity by more than 10% and decreased the activity of the peroxisomal fatty acid oxidation system by more than 10%. The effect was concentration-dependent. L-Arginine had the contrary effect. Combinations of L-arginine and N-nitro-L-arginine methyl ester hydrochloride compensated one another. If applied directly to the assays, S-nitroso-N-acetylpenicillamine and sodium nitroprusside inhibited catalase activity in a concentration-dependent manner. Sodium nitro-prusside had no effect on the peroxisomal beta-oxidation system unless cells were pretreated with N-nitro-L-arginine methyl ester overnight (50% inhibition). The results show a differential effect for the application of nitric oxide-effectors on fibroblast monolayers, cell suspensions and under assay conditions. Depending on the conditions of the incubation, nitric oxide applied to the cell monolayer at low doses acts as a second messenger in cells rather than as reactive oxygen species. Under assay conditions the effect of nitric oxide is more likely that of a reactive oxygen species because it inhibits all measured enzyme activities.