Fatty Acid Dehydrogenase Research Articles

Characterization of hydroxy fatty acid dehydrogenase involved in polyunsaturated fatty acid saturation metabolism in Lactobacillus plantarum AKU 1009a

Hydroxy fatty acid dehydrogenase, which is involved in polyunsaturated fatty acid saturation metabolism in Lactobacillus plantarum AKU 1009a, was cloned, expressed, purified, and characterized. The enzyme preferentially catalyzed NADH-dependent hydrogenation of oxo fatty acids over NAD+-dependent dehydrogenation of hydroxy fatty acids. In the dehydrogenation reaction, fatty acids with an internal hydroxy group such as 10-hydroxy-cis-12-octadecenoic acid, 12-hydroxy-cis-9-octadecenoic acid, and 13-hydroxy-cis-9-octadecenoic acid served as better substrates than those with α- or β-hydroxy groups such as 3-hydroxyoctadecanoic acid or 2-hydroxyeicosanoic acid. The apparent Km value for 10-hydroxy-cis-12-octadecenoic acid (HYA) was estimated to be 38μM with a kcat of 7.6×10−3s−1. The apparent Km value for 10-oxo-cis-12-octadecenoic acid (KetoA) was estimated to be 1.8μM with a kcat of 5.7×10−1s−1. In the hydrogenation reaction of KetoA, both (R)- and (S)-HYA were generated, indicating that the enzyme has low stereoselectivity. This is the first report of a dehydrogenase with a preference for fatty acids with an internal hydroxy group.

Anti-senescence effect of FatI gene in goat somatic cells.

The fatty acid dehydrogenase I (FatI) is able to express in mammalian cells and convert n-6 polyunsaturated fatty acids (PUFAs) to n-3 PUFAs. n-3 PUFA is an important component of the cell membrane and plays an important role in the prevention and control of a variety of human diseases. However, n-3 PUFAs cannot be endogenously synthesized by mammals because they lack the dehydrogenase that converts n-6 to n-3 PUFA. For the time being, gradually matured transgenic technology makes it possible to produce transgenic animals that are able to synthesize n-3 PUFAs by themselves. However, the transgenic technology itself may bring negative impacts. In this study, the eukaryotic expression vector pcDNA3.1-FatI was introduced into the genome of Boer goat fetal fibroblasts cultured in vitro, and the influence of biological characteristics of the fetal fibroblast was studied via overexpression of FatI. The results showed that the proliferation and apoptosis of cultured fetal fibroblast were not affected significantly by the overexpression of FatI using BrdU and TUNEL staining methods, respectively. Moreover, the overexpression of FatI significantly inhibited the senescence of somatic cells compared with enhanced green fluorescent protein (EGFP) transgenic cells (P < 0.01). Quantitative PCR revealed that the mRNA expression of P16 and P53 in the FatI transgenic cell group was significantly lower than that in the EGFP transgenic cell group (P < 0.01). In conclusion, the senescence of goat somatic cells was inhibited by the overexpression of the FatI gene.

Cloning and in vitro function analysis of codon‐optimized FatI gene

Currently, n-3 polyunsaturated fatty acids (n-3 PUFAs) have attracted great attention because of their biological significance to organisms. In addition, PUFAs show an obvious impact on prevention and treatment of various diseases. Because n-3 PUFAs cannot be endogenously synthesized by mammals, mammals have to rely on a dietary supplement for sufficient supply. The finding and application of the fatty acid dehydrogenase I (FatI) gene are expected to change the current situation because it can convert n-6 polyunsaturated fatty acids (n-6 PUFAs) to n-3 PUFAs. Meanwhile, the gradual maturation of transgenic technology makes it possible to produce transgenic animals that can synthesize n-3 PUFAs by themselves. In this study, the DNA coding sequence of FatI was synthesized by a chemical method after codon optimization according to the mammal's codon bias. The synthesized DNA sequence was introduced into Boer goat fetal fibroblasts by the constructed recombinant eukaryotic expression vector pcDNA3.1(+)-FatI. Boer goat fetal fibroblasts were transfected by electroporation, and the stable transfected cell lines were obtained by G418 selection. Genomic DNA PCR and Southern blot were applied to verify that the foreign gene FatI was integrated into the genome of the Boer goat fibroblasts. RT-PCR results showed the expression of FatI gene at the mRNA level. The fatty acid profile of cells carrying the FatI gene revealed an increase in total n-3 PUFAs (from 0.61 to 0.95), but a decrease in n-6 PUFAs (from 10.34 to 9.85), resulting in a remarkable increase in the n-3:n-6 ratio (from 0.059 to 0.096). The n-3:n-6 ratio had a 63.49 percent increase, which is a precursor of the response of n-3 desaturase activity of the FatI gene. The study may provide a practical tool for producing transgenic animals that can produce n-3 PUFAs by themselves, and we hope that the application will lay the foundation for animals producing n-3 PUFAs, which will benefit human nutrition and wellness.

Induction of time-dependent oxidative stress and related transcriptional effects of perfluorododecanoic acid in zebrafish liver

The effects of acute perfluorododecanoic acid (PFDoA) exposure on the induction of oxidative stress and alteration of mitochondrial gene expression were studied in the livers of female zebrafish ( Danio rerio). Female zebrafish were exposed to PFDoA via a single intraperitoneal injection (0, 20, 40, or 80 μg PFDoA/g body weight) and were then sacrificed 48 h, 96 h, or seven days post-PFDoA administration. PFDoA-treated fish exhibited histopathological liver damage, including swollen hepatocytes, vacuolar degeneration, and nuclei pycnosis. Glutathione (GSH) content and catalase (CAT) activity decreased significantly at 48 h post-injection while superoxide dismutase (SOD) activity was initially decreased at 48 h post-injection but was then elevated by seven days post-injection. The activity of glutathione peroxidase (GPx) increased at 48 h and seven days compared to control fish, although the increased level at seven days post-injection was decreased compared to the level at 48 h post-injection. Lipid peroxidation levels were increased at seven days post-injection, while no apparent induction was observed at 48 h or 96 h post-injection. The mRNA expression of medium-chain fatty acid dehydrogenase (MCAD) was induced, while the transcriptional expression of liver fatty acid binding protein (L-FABP), peroxisome proliferating activating receptor α (PPARα), carnitine palmitoyl-transferase I (CPT-I), uncoupling protein 2 (UCP-2), and Bcl-2 were significantly inhibited. Furthermore, the transcriptional expression of peroxisomal fatty acyl-CoA oxidase (ACOX), very long-chain acyl-CoA dehydrogenase (VLCAD), long-chain acyl-CoA dehydrogenase (LCAD) did not exhibit significant changes following PFDoA treatment. No significant changes were noted in the transcriptional expression of genes involved in mitochondrial respiratory chain and ATP synthesis, including cytochrome c oxidase subunit I (COXI), NADH dehydrogenase subunit I (NDI), and ATP synthase F0 subunit 6 (ATPo6). These results demonstrate that turbulence of fatty acid β-oxidation and oxidative stress responses were involved in the PFDoA-induced hepatotoxicity.

Microbial activity and phospholipid fatty acid pattern in long-term tannery waste-contaminated soil

We investigated the phospholipid fatty acid (PLFA) pattern and dehydrogenase activity (DHA) in soil samples from three sites (designated as low, medium, and high based on the level of chromium) in a long-term (25 years after last waste input) tannery waste-contaminated area rich in Cr. Soil samples, collected from different soil depths (0–100 cm), at each site were used in this study. In general, soil samples from all three contaminated sites had elevated pH, electrical conductivity, organic carbon (OC), total Cr, and hexavalent Cr [Cr(VI)]. The maximum total Cr concentration in surface soils (0–10 cm) at the highly contaminated site was 102 g kg −1, with 4.6 mg kg −1 present as the bioavailable water-soluble Cr. More than 50% of soluble Cr was in the form of Cr(VI) (2.7 mg kg −1). DHA (normalized to OC) was inhibited to a greater extent in soil samples from the highly contaminated site than in low- and medium-contaminated soil samples. PLFA analyses of surface soils indicated that there was a shift in PLFA patterns. PLFAs specific for bacteria (i15:0, a15:0, 15:0, i16:0, a17:0, and cy17:0) decreased significantly ( P<0.01) with an increase in Cr contamination. Among the bacterial PLFAs, 15:0, i16:0 and a17:0 had a significant negative correlation with contamination including bioavailable Cr(VI) in soil solution. To our knowledge, this is the first report of alterations in the PLFA profile in soils due to long-term tannery waste pollution.

Comparison of microbial and meiofaunal community analyses for determining impact of heavy metal contamination

The impact of long-term heavy metal contamination on soil communities was assessed by a number of methods. These included plate counts of culturable bacteria, community level physiological profiling (CLPP) by analysis of the utilization of multiple carbon sources in BIOLOG plates, community fatty acid methyl ester (C-FAME) profiling and dehydrogenase enzyme activity measurements. These approaches were complemented with microscopic assessments of the diversity of the nematode community. Samples from two sites with different histories of heavy-metal input were assessed. Major differences in microbial and meiofaunal parameters were observed both between and within the sites. There was a large degree of congruence between each of the microbiological approaches. In particular, one sample appeared to be distinguished by a reduction in culturable bacteria (especially pseudomonads), limited response to carbon sources in CLPP, and major differences in extracted fatty acid profiles. The use of multivariate analysis to examine the relationship between microbial and physicochemical measurements revealed that CLPP and plate counts were useful for determining the gross effect of metals on soil microbial communities, whereas proportions of metal-resistant bacteria and dehydrogenase activity differentiated between the two sites. Copper and zinc concentrations and pH all showed significant correlation with the microbial parameters. Nematode community structure was affected to a greater extent by soil pH than by metal content, but the within-site rankings were the same as those achieved for microbiological analyses. The use of these methods for field evaluation of the impact of industrial pollution may be possible provided care is taken when interpreting the data.

Perfusion characteristics of preserved canine kidneys subjected to warm ischaemia.

Canine kidneys were subjected to 0, 15 or 30 min of warm ischaemia followed by 24 hours preservation by perfusion. Changes in perfusate concentration of acid radicles, lactate, free fatty acid and lactice dehydrogenase were assessed at 1 hour and 24 hours. With the exception of LDH concentration at 1 hour, no single parameter was capable of detecting kidneys which were so damaged as to be non-life supporting.

SEX DIFFERENCE OF METABOLISM IN RAT LIVER AFTER TESTOSTERONE PROPIONATE ADMINISTRATION.

Effect of long-term administration of testosterone propionate (TP) on the function of liver was studied using the Wistar-King A strain rats with special references to the sex and age.1. RNA and nitrogen contents in the liver mitochondrial fraction of female rats exceeded that in male animals after 40 days of age at which sex maturation began. TP administration produced an increase in the RNA and nitrogen contents in both sexes, particularly in female rats by long-term TP administration.2. Succinic dehydrogenase activity in the mitochondrial fraction of the liver showed marked sex difference after 40 days of age. The activity in female animals was higher than in males. TP increased the activity of succinic dehydrogenase in male rats, whereas decreased in female rats.3. Fatty acid dehydrogenase activity showed a marked increase after 40 days of age, but no sex difference appeared. No effects was observed after TP treatment in both sexes.

Omega hydroxy fatty acid dehydrogenase

1. 1. A DPN-dependent dehydrogenase which catalyzes the oxidation of C 9, C 10, and C 11 omega hydroxy fatty acids was extracted from hog liver. The postulated product of the oxidation, omega oxo fatty acid, reversed the reaction in the presence of the purified enzyme. 2. 2. The 200-fold purified dehydrogenase was specific for fatty acid alcohols of the medium molecular weight (C 9–C 11). This system is not active with very low (C 1–C 4) or very high molecular weight (C 18) omega hydroxy acids or with a variety of the primary alcohols which do or do not have additional functional groups, i.e., amino, carboxy, or a second hydroxyl. Secondary fatty acid alcohols were also unaffected. 3. 3. As judged by the spectrophotometric assay, the omega fatty acid dehydrogenase is present also in extracts of heart and kidney of hog and the liver of sheep, rats, and horses. 4. 4. The possible relationship of this enzyme to omega oxidation is discussed briefly.

植物種子中の高級脂肪酸脱水素酵素

The extract of green soybeans had the action of the higher fatty acid dehydrogenase. This dehydrogenase took only the following fatty acids as the substrate: myristic, palmitic, stearic, oleic, linoleic, and alpha-hydroxy, 12-hydroxy, 9, 10-dihydroxy, and 9, 10, 12-trihydroxystearic, acids, and the optimum pH of this enzyme was 6.8 and the optimum temperature was between 35 and 40°. By the use of 2, 3, 5-triphenyltetrazolium chloride, as the hydrogen acceptor and the indicator, it was observed that the enzyme reaction proceeded linearly with time.

植物種子中の高級脂肪酸脱水素酵素

植物種子中の高級脂肪酸脱水素酵素

Catalysis of linoleate oxidation by pea lipoxidase

1. 1. Pea extracts contain an enzyme which does not possess fatty acid oxidase or fatty acid dehydrogenase activity, yet oxidizes linoleate but not oleate. Hence, the enzyme behaves like a true lipoxidase. 2. 2. The absorption spectrum of the oxidation products of pea lipoxidase catalysis shows a maximum at 280 mμ besides the usual chromophore at 233 mμ. Analysis of the linoleate products revealed the presence of monocarbonyls. Based on oxygen, the ϵ M value of pea lipoxidase-catalyzed linoleate oxidation was found to be 8000. Based on peroxides, the ϵ M value obtained was 27,300. These observations suggest that the hydroperoxides formed at 0 °C. are unstable and give rise to many other oxygen-consuming reactions. The peroxides per se are probably conjugated dienes. The implications of the above findings in relation to the production of “off odors” in underblanched frozen peas is discussed. 3. 3. The optimum pH of pea lipoxidase using ammonium linoleate as a substrate and the manometric method of assay was found to be 6.9. The lipoxidase-catalyzed linoleate oxidation was a maximum between 0 and 15 °C. 4. 4. The antioxidants nordihydroguaiaretic acid, propyl gallate, and α-tocopherol were found to strongly inhibit the pea lipoxidase-catalyzed oxidation of linoleate. 5. 5. An extensive series of experiments did not reveal the presence of any prosthetic group or coenzyme. These included inhibitor experiments, exhaustive dialysis against buffer of various pH values, and treatment with activated carbon or with intestinal phosphatase. 6. 6. On the hypothesis that linoleate could act as a coenzyme for lipoxidase, the activity of such a postulated enzyme-coenzyme system as a DPNH-oxidase, cytochrome c-oxidase, diaphorase, cysteine, and glutathione oxidase was tested. Results in all cases were negative. 7. 7. The possible role of lipoxidase in the formation of lipoidal plant films is discussed.

脂肪酸脱水素酵素に関する研究

豚肝から脂肪酸脱水素酵素を抽出してアポ酵素を作り,これにco-factorとしてCo-enzyme I, FAD,アデニール酸及びヒポキサンチンを添加してメチレン青の脱色力を見た.その結果 1) Co-enzyme Iは脂肪酸脱水素系には関与しない. 2) FAD,アデニール酸及びヒポキサンチンは共に助酵素と考えられるが,その作用条件は違う.すなわち a) FADは単独でも働くがATP,フマール酸があると強くなる. b) アデニール酸はフマール酸が無いと脱色が非常に遅れる. c) ヒポキサンチンは単独で強く働き, ATPもフマール酸も必要としない.

On the mechanism of enzyme action. XLVIII. Investigation of the fat of Fusarium lini Bolley by means of urea adducts

The usefulness of urea adducts for the fractionation of natural fatty acids has been demonstrated. Stearic acid has been isolated and identified from the fat of Fusarium lini Bolley by purification of the solid fraction obtained from the urea adduct of its fatty acids. The conversion of stearic acid to fatty material with a higher iodine value has provided further evidence for the enzymatic formation of unsaturated fatty acids in Fusarium lini Bolley by means of the action of a fatty acid dehydrogenase system.

Interrelationship between natural pigments and the mechanism of conversion of carbohydrate into fat.

PARTICIPATION of Fusarium pigments in the conversion of carbohydrate into fat was indicated when it was found that the greatest rise in fat production by Fusarium solani D2 purple was paralleled by an increment in pigment formation; the fat content of Fusarium lycopersici obtained pigmented on Raulin–Thom medium increased with rising sugar concentration of the medium, while the reverse was true when the mould was grown unpigmented on Czapek-Dox medium; Fusarium lini Bolley grew unpigmented in both media and showed no change in fat content1. The function of these pigments was clearly indicated by experiments in which solanione was added to the media of growing Fusaria. The non-pigment formers produced a fat of greater desaturation under these conditions, whereas the fat formed in the pigmented moulds remained unaffected2. Evidence has been presented previously correlating the action of the fatty acid dehydrogenase system of Fusaria3 with that of the pigments so characteristic of this genus. Increased desaturation of the fats of the non-pigmented Fusarium lini Bolley was also caused by naphthoquinones chemically related to solanione4.

Higher Fatty Acid Dehydrogenase of Mammalian Liver

AN enzyme dehydrogenating higher fatty acids has been described by several workers, including Lang and Mayer1 and Annau et al.2 Lang and Mayer showed that adenylic acid activated such a system in rat liver, while Annau et al. showed that hypoxanthine similarly activated a fatty acid dehydrogenase of ox liver.

Fatty acid dehydrogenase in adipose tissue

Research Article| April 01 1943 Fatty acid dehydrogenase in adipose tissue B. Shapiro; B. Shapiro 1Department of Pathological Physiology, The Hebrew University, Jerusalem Search for other works by this author on: This Site PubMed Google Scholar E. Wertheimer E. Wertheimer 1Department of Pathological Physiology, The Hebrew University, Jerusalem Search for other works by this author on: This Site PubMed Google Scholar Biochem J (1943) 37 (1): 102–104. https://doi.org/10.1042/bj0370102 Views Icon Views Article contents Figures & tables Video Audio Supplementary Data Peer Review Share Icon Share Facebook Twitter LinkedIn MailTo Cite Icon Cite Get Permissions Citation B. Shapiro, E. Wertheimer; Fatty acid dehydrogenase in adipose tissue. Biochem J 1 April 1943; 37 (1): 102–104. doi: https://doi.org/10.1042/bj0370102 Download citation file: Ris (Zotero) Reference Manager EasyBib Bookends Mendeley Papers EndNote RefWorks BibTex toolbar search Search Dropdown Menu toolbar search search input Search input auto suggest filter your search All ContentAll JournalsBiochemical Journal Search Advanced Search This content is only available as a PDF. © 1943 CAMBRIDGE UNIVERSITY PRESS1943 Article PDF first page preview Close Modal You do not currently have access to this content.