20 results on '"Ogawa, Jun"'
Search Results
2. Characterization of ω3 fatty acid desaturases from oomycetes and their application toward eicosapentaenoic acid production in Mortierella alpina.
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Mo, Brian K H, Ando, Akinori, Nakatsuji, Ryohei, Okuda, Tomoyo, Takemoto, Yuki, Ikemoto, Hiroyuki, Kikukawa, Hiroshi, Sakamoto, Takaiku, Sakuradani, Eiji, and Ogawa, Jun
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FATTY acid desaturase ,UNSATURATED fatty acids ,EICOSAPENTAENOIC acid ,MORTIERELLA ,OOMYCETES ,ARACHIDONIC acid ,FATTY acids - Abstract
ω3 Polyunsaturated fatty acids are currently obtained mainly from fisheries; thus, sustainable alternative sources such as oleaginous microorganisms are required. Here, we describe the isolation, characterization, and application of 3 novel ω3 desaturases with ω3 polyunsaturated fatty acid–producing activity at ordinary temperatures (28 °C). First, we selected Pythium sulcatum and Plectospira myriandra after screening for oomycetes with high eicosapentaenoic acid/arachidonic acid ratios and isolated the genes psulω3 and pmd17 , respectively, which encode ω3 desaturases. Subsequent characterization showed that PSULω3 exhibited ω3 desaturase activity on both C18 and C20 ω6 polyunsaturated fatty acids while PMD17 exhibited ω3 desaturase activity exclusively on C20 ω6 polyunsaturated fatty acids. Expression of psulω3 and pmd17 in the arachidonic acid–producer Mortierella alpina resulted in transformants that produced eicosapentaenoic acid/total fatty acid values of 38% and 40%, respectively, at ordinary temperatures. These ω3 desaturases should facilitate the construction of sustainable ω3 polyunsaturated fatty acid sources. [ABSTRACT FROM AUTHOR]
- Published
- 2021
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3. Antimicrobial function of the polyunsaturated fatty acid KetoC in an experimental model of periodontitis.
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Sulijaya, Benso, Yamada‐Hara, Miki, Yokoji‐Takeuchi, Mai, Matsuda‐Matsukawa, Yumi, Yamazaki, Kyoko, Matsugishi, Aoi, Tsuzuno, Takahiro, Sato, Keisuke, Aoki‐Nonaka, Yukari, Takahashi, Naoki, Kishino, Shigenobu, Ogawa, Jun, Tabeta, Koichi, Yamazaki, Kazuhisa, Yamada-Hara, Miki, Yokoji-Takeuchi, Mai, Matsuda-Matsukawa, Yumi, and Aoki-Nonaka, Yukari
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UNSATURATED fatty acids ,PERIODONTITIS ,ANTI-infective agents ,METABOLITES ,LABORATORY mice ,POLYMERASE chain reaction ,BIOLOGICAL models ,RESEARCH ,ANIMAL experimentation ,RESEARCH methodology ,GRAM-negative anaerobic bacteria ,EVALUATION research ,MEDICAL cooperation ,COMPARATIVE studies ,RESEARCH funding ,MICE ,ANTIBIOTICS - Abstract
Background: The bioactive metabolite KetoC, generated by intestinal bacteria, exerts various beneficial effects. Nevertheless, its function in the pathogenesis of periodontitis remains unclear. Here, we investigated the effect of KetoC in a mouse model of periodontitis and explored the underlying mechanism.Methods: Thirty-one 8-week-old male C57BL/6N mice were randomly divided into four groups (non-ligation, non-ligation + KetoC, ligation + Porphyromonas gingivalis, and ligation + P. gingivalis + KetoC) (n = 7/8 mice/group) and given a daily oral gavage of KetoC (15 mg/mL) or vehicle for 2 weeks. To induce periodontitis, a 5-0 silk ligature was placed on the maxillary left second molar on day 7, and P. gingivalis W83 (109 colony-forming unit [CFU]) was administered orally every 3 days. On day 14, all mice were euthanized. Alveolar bone destruction was determined from the level of the cemento-enamel junction to the alveolar bone crest. Moreover, bone loss level was confirmed from gingival tissue sections stained with hematoxylin and eosin. The presence of P. gingivalis was quantified using real-time polymerase chain reaction. In vitro, the bacteriostatic and bactericidal effects of KetoC were assessed by analyzing its suppressive activity on the proliferation of P. gingivalis and using a live/dead bacterial staining kit, respectively. A double-bond-deficient metabolite (KetoB) was then used to investigate the importance of double-bond structure in the antimicrobial activity of KetoC on P. gingivalis.Results: In vivo, KetoC attenuated alveolar bone destruction and suppressed P. gingivalis in the periodontitis group. In vitro, KetoC (but not KetoB) downregulated the proliferation and viability of P. gingivalis in a dose-dependent manner.Conclusions: KetoC reduced alveolar bone destruction in a periodontitis model via its antimicrobial function. Therefore, this bioactive metabolite may be valuable in clinical applications to support periodontal therapy. [ABSTRACT FROM AUTHOR]- Published
- 2019
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4. Supplemental feeding of a gut microbial metabolite of linoleic acid, 10-hydroxy-cis-12-octadecenoic acid, alleviates spontaneous atopic dermatitis and modulates intestinal microbiota in NC/nga mice.
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Kaikiri, Hiroko, Miyamoto, Junki, Kawakami, Takahiro, Park, Si-Bum, Kitamura, Nahoko, Kishino, Shigenobu, Yonejima, Yasunori, Hisa, Keiko, Watanabe, Jun, Ogita, Tasuku, Ogawa, Jun, Tanabe, Soichi, and Suzuki, Takuya
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ATOPIC dermatitis ,MICROBIAL metabolites ,GUT microbiome ,DIETARY supplements ,LINOLEIC acid ,OLEIC acid ,LABORATORY mice ,ANIMAL behavior ,ANIMAL experimentation ,CYTOKINES ,DIET ,FECES ,FOOD ,GENES ,IMMUNOGLOBULINS ,INFLAMMATION ,MICE ,MOLECULAR structure ,POLYMERASE chain reaction ,UNSATURATED fatty acids ,REVERSE transcriptase polymerase chain reaction - Abstract
The present study investigated the antiallergic and anti-inflammatory effects of 10-hydroxy-cis-12-octadecenoic acid (HYA), a novel gut microbial metabolite of linoleic acid, in NC/Nga mice, a model of atopic dermatitis (AD). Feeding HYA decreased the plasma immunoglobulin E level and skin infiltration of mast cells with a concomitant decrease in dermatitis score. HYA feeding decreased TNF-α and increased claudin-1, a tight junction protein, levels in the mouse skin. Cytokine expression levels in the skin and intestinal Peyer's patches cells suggested that HYA improved the Th1/Th2 balance in mice. Immunoglobulin A concentration in the feces of the HYA-fed mice was approximately four times higher than that in the control mice. Finally, denaturing gradient gel electrophoresis of the PCR-amplified 16 S rRNA gene of fecal microbes indicated the modification of microbiota by HYA. Taken together, the alterations in the intestinal microbiota might be, at least in part, associated with the antiallergic effect of HYA. [ABSTRACT FROM AUTHOR]
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- 2017
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5. Production of dicarboxylic acids from novel unsaturated fatty acids by laccase-catalyzed oxidative cleavage.
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Takeuchi, Michiki, Kishino, Shigenobu, Park, Si-Bum, Kitamura, Nahoko, Watanabe, Hiroko, Saika, Azusa, Hibi, Makoto, Yokozeki, Kenzo, and Ogawa, Jun
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DICARBOXYLIC acids ,UNSATURATED fatty acids ,LACCASE - Abstract
The establishment of renewable biofuel and chemical production is desirable because of global warming and the exhaustion of petroleum reserves. Sebacic acid (decanedioic acid), the material of 6,10-nylon, is produced from ricinoleic acid, a carbon-neutral material, but the process is not eco-friendly because of its energy requirements. Laccase-catalyzing oxidative cleavage of fatty acid was applied to the production of dicarboxylic acids using hydroxy and oxo fatty acids involved in the saturation metabolism of unsaturated fatty acids inLactobacillus plantarumas substrates. Hydroxy or oxo fatty acids with a functional group near the carbon–carbon double bond were cleaved at the carbon–carbon double bond, hydroxy group, or carbonyl group by laccase and transformed into dicarboxylic acids. After 8 h, 0.58 mM of sebacic acid was produced from 1.6 mM of 10-oxo-cis-12,cis-15-octadecadienoic acid (αKetoA) with a conversion rate of 35% (mol/mol). This laccase-catalyzed enzymatic process is a promising method to produce dicarboxylic acids from biomass-derived fatty acids. Oxidative cleavage of oxo unsaturated fatty acid into dicarboxylic acid by laccase. [ABSTRACT FROM PUBLISHER]
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- 2016
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6. Metabolic diversity in biohydrogenation of polyunsaturated fatty acids by lactic acid bacteria involving conjugated fatty acid production.
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Kishino, Shigenobu, Ogawa, Jun, Yokozeki, Kenzo, and Shimizu, Sakayu
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LACTIC acid bacteria , *UNSATURATED fatty acids , *LINOLEIC acid , *OLEIC acid , *LACTIC acid , *FATTY acids - Abstract
Lactobacillus plantarum AKU 1009a effectively transforms linoleic acid to conjugated linoleic acids of cis-9, trans-11-octadecadienoic acid (18:2) and trans-9, trans-11–18:2. The transformation of various polyunsaturated fatty acids by washed cells of L. plantarum AKU 1009a was investigated. Besides linoleic acid, α-linolenic acid [ cis-9, cis-12, cis-15-octadecatrienoic acid (18:3)], γ-linolenic acid ( cis-6, cis-9, cis-12–18:3), columbinic acid ( trans-5, cis-9, cis-12–18:3), and stearidonic acid [ cis-6, cis-9, cis-12, cis-15-octadecatetraenoic acid (18:4)] were found to be transformed. The fatty acids transformed by the strain had the common structure of a C18 fatty acid with the cis-9, cis-12 diene system. Three major fatty acids were produced from α-linolenic acid, which were identified as cis-9, trans-11, cis-15–18:3, trans-9, trans-11, cis-15–18:3, and trans-10, cis-15–18:2. Four major fatty acids were produced from γ-linolenic acid, which were identified as cis-6, cis-9, trans-11–18:3, cis-6, trans-9, trans-11–18:3, cis-6, trans-10–18:2, and trans-10-octadecenoic acid. The strain transformed the cis-9, cis-12 diene system of C18 fatty acids into conjugated diene systems of cis-9, trans-11 and trans-9, trans-11. These conjugated dienes were further saturated into the trans-10 monoene system by the strain. The results provide valuable information for understanding the pathway of biohydrogenation by anaerobic bacteria and for establishing microbial processes for the practical production of conjugated fatty acids, especially those produced from α-linolenic acid and γ-linolenic acid. [ABSTRACT FROM AUTHOR]
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- 2009
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7. Improved production of various polyunsaturated fatty acids through filamentous fungus Mortierella alpina breeding.
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Sakuradani, Eiji, Ando, Akinori, Ogawa, Jun, and Shimizu, Sakayu
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UNSATURATED fatty acids ,MORTIERELLA ,BIOSYNTHESIS ,ARACHIDONIC acid ,FATTY acids ,FUNGI - Abstract
Studies on the application of functional lipids such as polyunsaturated fatty acids (PUFAs) have proceeded in various fields regarding health and dietary requirements in a search for novel and rich sources. Filamentous fungus Mortierella alpina 1S-4 produces triacylglycerols rich in arachidonic acid, ones reaching 20 g/L and containing 30–70% arachidonic acid as to the total fatty acids. Mutants derived from M. alpina 1S-4, defective in Δ5 and Δ6 desaturases, accumulate triacylglycerols rich in unique PUFAs, i.e., dihomo-γ-linolenic acid and Mead acid, respectively. Furthermore, various mutants derived from M. alpina 1S-4 have led to the production of oils containing n−1, n−3, n−4, n−6, n−7, and n−9 PUFAs. A variety of genes encoding fatty acid desaturases and elongases involved in PUFA biosynthesis in M. alpina 1S-4 has been isolated and characterized. Molecular breeding of M. alpina strains by means of manipulation of these genes facilitates improvement of PUFA productivity and elucidation of the functions of enzymes involved in PUFA biosynthesis. [ABSTRACT FROM AUTHOR]
- Published
- 2009
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8. Fatty Acid Desaturation and Elongation Reactions of Trichoderma sp. 1-OH-2-3.
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Ando, Akinori, Ogawa, Jun, Kishino, Shigenobu, Ito, Taiyo, Shirasaka, Norifumi, Sakuradani, Eiji, Yokozeki, Kenzo, and Shimizu, Sakayu
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UNSATURATED fatty acids ,TRICHODERMA ,PALMITIC acid ,STEARIC acid ,OLEIC acid ,LINOLEIC acid ,LINOLENIC acids ,STRAINS & stresses (Mechanics) - Abstract
The fatty acid desaturation and elongation reactions catalyzed by Trichoderma sp. 1-OH-2-3 were investigated. This strain converted palmitic acid (16:0) mainly to stearic acid (18:0), and further to oleic acid ( c9-18:1), linoleic acid ( c9, c12-18:2), and α-linolenic acid ( c9, c12, c15-18:3) through elongation, and Δ9, Δ12, and Δ15 desaturation reactions, respectively. Palmitoleic acid ( c9-16:1) and cis-9, cis-12-hexadecadienoic acid were also produced from 16:0 by the strain. This strain converted n-tridecanoic acid (13:0) to cis-9-heptadecenoic acid and further to cis-9, cis-12-heptadecadienoic acid through elongation, and Δ9 and Δ12 desaturation reactions, respectively. trans-Vaccenic acid ( t11-18:1) and trans-12-octadecenoic acid ( t12-18:1) were desaturated by the strain through Δ9 desaturation. The products derived from t11-18:1 were identified as the conjugated linoleic acids (CLAs) of cis-9, trans-11-octadecadienoic acid and trans-9, trans-11-octadecadienoic acid. The product derived from t12-18:1 was identified as cis-9, trans-12-octadecadienoic acid. cis-6, cis-9-Octadecadienoic acid was desaturated to cis-6, cis-9, cis-12-octadecatrienoic acid by this strain through Δ12 desaturation. The broad substrate specificity of the elongation, and Δ9 and Δ12 desaturation reactions of the strain is useful for fatty acid biotransformation. [ABSTRACT FROM AUTHOR]
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- 2009
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9. Transformation of an oleaginous zygomycete Mortierella alpina 1S-4 with the carboxin resistance gene conferred by mutation of the iron–sulfur subunit of succinate dehydrogenase.
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Ando, Akinori, Sakuradani, Eiji, Horinaka, Kota, Ogawa, Jun, and Shimizu, Sakayu
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SUCCINATE dehydrogenase ,BACTERIAL transformation ,UNSATURATED fatty acids ,GLUCURONIDASE ,GENETIC mutation ,AMINO acid sequence - Abstract
The sdhB gene encoding an iron–sulfur (Ip) subunit of succinate dehydrogenase (SDH, EC 1.3.99.1) complex was cloned from Mortierella alpina 1S-4. The deduced amino acid sequence of SdhB from M. alpina 1S-4 showed high similarity to those of SdhB from other organisms. The mutated sdhB ( CBXB) gene encodes a modified SdhB with an amino-acid substitution (a highly conserved histidine residue within the third cysteine-rich cluster of SdhB replaced by a leucine residue) and is known to confer carboxin resistance. We succeeded in transforming M. alpina 1S-4 by using the CBXB gene as a selectable marker gene and expressing the heterologous uidA gene encoding β-glucuronidase of Escherichia coli. Moreover, transformation efficiency was up to 40–50 transformants per 4.0 × 10
8 spores. This carboxin-transformation system, characterized by marginal background growth and mitotic stability in M. alpina 1S-4, is considered to be widely useful for the wild strain, M. alpina 1S-4, and various derivative mutants without laborious preparation of auxotrophic mutants as a host strain. [ABSTRACT FROM AUTHOR]- Published
- 2009
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10. Gut microbiota confers host resistance to obesity by metabolizing dietary polyunsaturated fatty acids.
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Miyamoto, Junki, Igarashi, Miki, Watanabe, Keita, Karaki, Shin-ichiro, Mukouyama, Hiromi, Kishino, Shigenobu, Li, Xuan, Ichimura, Atsuhiko, Irie, Junichiro, Sugimoto, Yukihiko, Mizutani, Tetsuya, Sugawara, Tatsuya, Miki, Takashi, Ogawa, Jun, Drucker, Daniel J., Arita, Makoto, Itoh, Hiroshi, and Kimura, Ikuo
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UNSATURATED fatty acids ,GUT microbiome ,NATURAL immunity ,LINOLEIC acid ,OMEGA-6 fatty acids ,FREE fatty acids ,MICROBIAL metabolites - Abstract
Gut microbiota mediates the effects of diet, thereby modifying host metabolism and the incidence of metabolic disorders. Increased consumption of omega-6 polyunsaturated fatty acid (PUFA) that is abundant in Western diet contributes to obesity and related diseases. Although gut-microbiota-related metabolic pathways of dietary PUFAs were recently elucidated, the effects on host physiological function remain unclear. Here, we demonstrate that gut microbiota confers host resistance to high-fat diet (HFD)-induced obesity by modulating dietary PUFAs metabolism. Supplementation of 10-hydroxy-cis-12-octadecenoic acid (HYA), an initial linoleic acid-related gut-microbial metabolite, attenuates HFD-induced obesity in mice without eliciting arachidonic acid-mediated adipose inflammation and by improving metabolic condition via free fatty acid receptors. Moreover, Lactobacillus-colonized mice show similar effects with elevated HYA levels. Our findings illustrate the interplay between gut microbiota and host energy metabolism via the metabolites of dietary omega-6-FAs thereby shedding light on the prevention and treatment of metabolic disorders by targeting gut microbial metabolites. The gut microbiome is an important regulator of metabolic health. Here the authors show that intestinal bacteria metabolize dietary linoleic acid to 10-hydroxy-cis-12-octadecenoic acid (HYA) which confers host resistance to high fat diet-induced obesity in mice. [ABSTRACT FROM AUTHOR]
- Published
- 2019
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11. Mead acid production by disruption of Δ12-desaturase gene in Mortierella alpina 1S-4.
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Kikukawa, Hiroshi, Ando, Akinori, Hannya, Asuka, Farida Asras, Mohd Fazli, Okuda, Tomoyo, Sakamoto, Takaiku, Hara, Kiyotaka Y., Sakuradani, Eiji, and Ogawa, Jun
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UNSATURATED fatty acids , *MORTIERELLA , *LINOLEIC acid , *OLEIC acid , *FATTY acids , *ACIDS , *LINOLENIC acids , *PALMITIC acid - Abstract
Mead acid (MA; 20:3ω9) is one of the ω9 series of polyunsaturated fatty acids (PUFAs). MA is used to inhibit the inflammation of joints and is applied to the medicinal or health food field. We aimed to construct MA-producing strains with disruption of the Δ12-desaturase gene (Δ1 2ds) via an efficient gene-targeting system using the lig4 -disrupted strain of Mortierella alpina 1S-4 as the host. The transformants showed a unique fatty acid composition that only comprised ω9-PUFAs and saturated fatty acids, while ω6-and ω3-PUFAs were not detected, and the total composition of ω9-PUFAs, including oleic acid (18:1ω9), 18:2ω9, 20:1ω9, 20:2ω9, and MA, was up to 68.4% of the total fatty acids. The MA production in the Δ1 2ds -disruptant reached 0.10 g/L (8.5%), which exceeded 0.050 g/L (4.6%) in the conventional Δ1 2ds -defective mutant JT-180. [ABSTRACT FROM AUTHOR]
- Published
- 2023
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12. Isolation and characterization of the ω3-docosapentaenoic acid-producing microorganism Aurantiochytrium sp. T7.
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Wu, Chang-Yu, Okuda, Tomoyo, Ando, Akinori, Hatano, Ayami, Kikukawa, Hiroshi, and Ogawa, Jun
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FATTY acids , *UNSATURATED fatty acids , *FUNCTIONAL analysis - Abstract
ω3-Docosapentaenoic acid (ω3-DPA), an ω3-polyunsaturated fatty acid (ω3-PUFA), is expected to have beneficial physiological functions to humans; however, because of its rarity in nature, it has not been fully analyzed. We isolated an ω3-DPA producing microorganism strain T7 from brackish areas in Japan. Although most oleaginous microorganisms rarely accumulate ω3-DPA (<5% of total lipid), strain T7 accumulated ω3-DPA with more than 20% of total fatty acids. The strain T7 was identified as a related species of Aurantiochytrium. In Aurantiochytrium sp. T7, ω3-DPA production reached 164 mg/L culture broth, and the ω3-DPA content reached 23.5% of the total fatty acids when cultivated in a medium containing 2% glucose as the carbon source and 1% yeast extract as the nitrogen source, with a salinity equivalent to 50% of that of seawater and a pH in the acidic range (pH < 5.5). Aurantiochytrium sp. T7 is a promising producer of high-purity ω3-DPA containing-lipid for the functional analysis of ω3-DPA whose physiological function has hardly been elucidated, and a useful strain for investigating the novel metabolic pathway of fatty acids. [ABSTRACT FROM AUTHOR]
- Published
- 2022
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13. The anti-inflammatory effect of the gut lactic acid bacteria-generated metabolite 10-oxo-cis-6,trans-11-octadecadienoic acid on monocytes.
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Hagiwara, Shuhei, Nagata, Kazuki, Kasakura, Kazumi, Sakata, Fumiya, Kishino, Shigenobu, Ogawa, Jun, Yashiro, Takuya, and Nishiyama, Chiharu
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LACTIC acid , *G protein coupled receptors , *UNSATURATED fatty acids , *BACTERIAL metabolites , *LACTIC acid bacteria , *MICROBIAL metabolites , *LIPOPOLYSACCHARIDES - Abstract
We evaluated the effect of gut bacterial metabolites of polyunsaturated fatty acids on inflammation and found that 10-oxo- cis -6, trans -11-octadecadienoic acid (γKetoC) strikingly suppressed LPS-induced IL-6 release from bone marrow-derived macrophages (BMMs), which was accompanied by reduced mRNA expression of Il6 , TNF , and Il1b. γKetoC decreased the cAMP concentration in BMMs, suggesting that γKetoC stimulated G protein-coupled receptors. A Gq agonist significantly suppressed LPS-induced IL-6 expression in BMMs, whereas a Gi inhibitor partially abrogated γKetoC-mediated IL-6 suppression. Cytosolic Ca2+ was markedly increased by γKetoC, which was partly but not fully abrogated by an ion channel inhibitor. Taken together, these data suggest that γKetoC suppresses inflammatory cytokine expression in macrophages primarily through Gq and partially through Gi. γKetoC suppressed osteoclast development and IL-6 expression in synovial fibroblasts from rheumatoid arthritis (RA) patients, suggesting the beneficial effect of γKetoC on the prevention or treatment of RA. • A gut lactic acid bacteria metabolite, γKetoC, suppresses activation of macrophages. • γKetoC decreases inflammatory cytokine release from macrophages via GPCR. • γKetoC inhibits osteoclast development and IL-6 expression in synovial fibroblasts. [ABSTRACT FROM AUTHOR]
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- 2020
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14. Arachidonic acid production by the oleaginous fungus Mortierella alpina 1S-4: A review.
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Kikukawa, Hiroshi, Sakuradani, Eiji, Ando, Akinori, Shimizu, Sakayu, and Ogawa, Jun
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ARACHIDONIC acid , *MORTIERELLA , *TRIGLYCERIDES , *UNSATURATED fatty acids , *BIOSYNTHESIS - Abstract
The filamentous fungus Mortierella alpina 1S-4 is capable of accumulating a large amount of triacylglycerol containing C20 polyunsaturated fatty acids (PUFAs). Indeed, triacylglycerol production by M. alpina 1S-4 can reach 20 g/L of culture broth, and the critical cellular signaling and structural PUFA arachidonic acid (ARA) comprises 30%–70% of the total fatty acid. The demonstrated health benefits of functional PUFAs have in turn encouraged the search for rich sources of these compounds, including fungal strains showing enhanced production of specific PUFAs. Screening for mutants and targeted gene manipulation of M. alpina 1S-4 have elucidated the functions of various enzymes involved in PUFA biosynthesis and established lines with improved PUFA productivity. In some cases, these strains have been used for indistrial-scale production of PUFAs, including ARA. In this review, we described practical ARA production through mutant breeding, functional analyses of genes encoding enzymes involved in PUFA biosynthesis, and recent advances in the production of specific PUFAs through molecular breeding of M. alpina 1S-4. [ABSTRACT FROM AUTHOR]
- Published
- 2018
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15. Disruption of lig4 improves gene targeting efficiency in the oleaginous fungus Mortierella alpina 1S-4.
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Kikukawa, Hiroshi, Sakuradani, Eiji, Ando, Akinori, Okuda, Tomoyo, Ochiai, Misa, Shimizu, Sakayu, and Ogawa, Jun
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UNSATURATED fatty acids , *GENE targeting , *DNA ligases , *FUNGAL spores , *MORTIERELLA , *DNA repair , *GENE replacement - Abstract
The oil-producing zygomycete Mortierella alpina 1S-4 is known to accumulate beneficial polyunsaturated fatty acids. We identified the lig4 gene that encodes for a DNA ligase 4 homolog, which functions to repair double strand breaks by non-homologous end joining. We disrupted the lig4 gene to improve the gene targeting efficiency in M. alpina . The M. alpina 1S-4 Δlig4 strains showed no defect in vegetative growth, formation of spores, and fatty acid production, but exhibited high sensitivity to methyl methansulfonate, an agent that causes DNA double-strand breaks. Importantly, gene replacement of ura5 marker by CBXB marker occurred in 67% of Δlig4 strains and the gene targeting efficiency was 21-fold greater than that observed in disruption of the lig4 gene in the M. alpina 1S-4 host strain. Further metabolic engineering of the Δlig4 strains is expected to result in strains that produce higher levels of rare and beneficial polyunsaturated fatty acids and contribute to basic research on the zygomycete. [ABSTRACT FROM AUTHOR]
- Published
- 2015
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16. Characterization of hydroxy fatty acid dehydrogenase involved in polyunsaturated fatty acid saturation metabolism in Lactobacillus plantarum AKU 1009a.
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Takeuchi, Michiki, Kishino, Shigenobu, Park, Si-Bum, Kitamura, Nahoko, and Ogawa, Jun
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UNSATURATED fatty acids , *HYDROXY acids , *DEHYDROGENASES , *BACTERIAL metabolism , *SATURATION (Chemistry) , *LACTOBACILLUS plantarum , *DEHYDROGENATION - Abstract
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 K m value for 10-hydroxy- cis -12-octadecenoic acid (HYA) was estimated to be 38 μM with a k cat of 7.6 × 10 −3 s −1 . The apparent K m value for 10-oxo- cis -12-octadecenoic acid (KetoA) was estimated to be 1.8 μM with a k cat of 5.7 × 10 −1 s −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. [ABSTRACT FROM AUTHOR]
- Published
- 2015
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17. Characterization of the linoleic acid Δ9 hydratase catalyzing the first step of polyunsaturated fatty acid saturation metabolism in Lactobacillus plantarum AKU 1009a.
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Takeuchi, Michiki, Kishino, Shigenobu, Hirata, Akiko, Park, Si-Bum, Kitamura, Nahoko, and Ogawa, Jun
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LINOLEIC acid , *LACTOBACILLUS plantarum , *HYDRATASES , *UNSATURATED fatty acids , *NAD (Coenzyme) - Abstract
Linoleic acid Δ9 hydratase, which is involved in linoleic acid saturation metabolism of Lactobacillus plantarum AKU 1009a, was cloned, expressed as a his-tagged recombinant enzyme, purified with an affinity column, and characterized. The enzyme required FAD as a cofactor and its activity was enhanced by NADH. The maximal activities for the hydration of linoleic acid and for the dehydration of 10-hydroxy- cis -12-octadecenoic acid (HYA) were observed at 37 °C in buffer at pH 5.5 containing 0.5 M NaCl. Free C16 and C18 fatty acids with cis -9 double bonds and 10-hydroxy fatty acids served as substrates for the hydration and dehydration reactions, respectively. The apparent K m value for linoleic acid was estimated to be 92 μM, with a k cat of 2.6∙10 −2 s −1 and a Hill factor of 3.3. The apparent K m value for HYA was estimated to be 98 μM, with a k cat of 1.2∙10 −3 s −1 . [ABSTRACT FROM AUTHOR]
- Published
- 2015
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18. 10-oxo-12(Z)-octadecenoic acid, a linoleic acid metabolite produced by gut lactic acid bacteria, potently activates PPARγ and stimulates adipogenesis.
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Goto, Tsuyoshi, Kim, Young-Il, Furuzono, Tomoya, Takahashi, Nobuyuki, Yamakuni, Kanae, Yang, Ha-Eun, Li, Yongjia, Ohue, Ryuji, Nomura, Wataru, Sugawara, Tatsuya, Yu, Rina, Kitamura, Nahoko, Park, Si-Bum, Kishino, Shigenobu, Ogawa, Jun, and Kawada, Teruo
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OLEIC acid , *LINOLEIC acid , *METABOLITES , *LACTIC acid bacteria , *ADIPOGENESIS , *UNSATURATED fatty acids - Abstract
Our previous study has shown that gut lactic acid bacteria generate various kinds of fatty acids from polyunsaturated fatty acids such as linoleic acid (LA). In this study, we investigated the effects of LA and LA-derived fatty acids on the activation of peroxisome proliferator-activated receptors (PPARs) which regulate whole-body energy metabolism. None of the fatty acids activated PPARδ, whereas almost all activated PPARα in luciferase assays. Two fatty acids potently activated PPARγ, a master regulator of adipocyte differentiation, with 10-oxo-12( Z )-octadecenoic acid (KetoA) having the most potency. In 3T3-L1 cells, KetoA induced adipocyte differentiation via the activation of PPARγ, and increased adiponectin production and insulin-stimulated glucose uptake. These findings suggest that fatty acids, including KetoA, generated in gut by lactic acid bacteria may be involved in the regulation of host energy metabolism. [ABSTRACT FROM AUTHOR]
- Published
- 2015
- Full Text
- View/download PDF
19. Characterization of a trifunctional fatty acid desaturase from oleaginous filamentous fungus Mortierella alpina 1S-4 using a yeast expression system.
- Author
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Kikukawa, Hiroshi, Sakuradani, Eiji, Kishino, Shigenobu, Park, Si-Bum, Ando, Akinori, Shima, Jun, Ochiai, Misa, Shimizu, Sakayu, and Ogawa, Jun
- Subjects
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FATTY acid desaturase , *FILAMENTOUS fungi , *MORTIERELLA , *YEAST , *BIOSYNTHESIS , *UNSATURATED fatty acids , *ENZYME kinetics - Abstract
A ω3-fatty acid desaturase gene (maw3) which is involved in biosynthesis of n-3 polyunsaturated fatty acids (PUFAs) was previously isolated from Mortierella alpina 1S-4. In this report, we investigated the products of MAW3 catalyzing reaction with endogenous and exogenous fatty acids in the yeast transformant. Two unusual fatty acids de novo synthesized in the yeast transformant expressing maw3 gene were identified as n-4 hexadecadienoic acid (16:29cis,12cis ) and n-1 hexadecatrienoic acid (16:39cis,12cis,15) by GC–MS and 1H NMR analyses. In addition to the desaturation activity at the ω3-position for 18- and 20-carbon PUFAs, MAW3 in the yeast transformant inserted a double bond at Δ12-position of endogenous palmitoleic acid (16:19cis ) and further at Δ15-position of the resulting 16:29cis,12cis to result in the formation of 16:39cis,12cis,15 leading to a bifunctional Δ12/Δ15-desaturase for 16-carbon fatty acids. Moreover, we evaluated the activity of MAW3 in the yeast transformant under different temperatures. The MAW3 did not have desaturation activities in M. alpina 1S-4 at 28°C but it had in the yeast transformant for various fatty acids. The MAW3 was demonstrated to be a trifunctional Δ12/Δ15/ω3-desaturase, exhibiting Δ12-desaturation for 16:19cis , Δ15-desaturation for 16- and 18-carbon fatty acids that had a preexisting cis-double bond at Δ12 position, and ω3-desaturation for 20-carbon fatty acids having that at Δ14-position. It is the first report that the fatty acid desaturase (MAW3) is shown to have Δ12- and Δ15-desaturation activities for a 16-carbon fatty acid, in addition to its major function, ω3-desaturation activity. [ABSTRACT FROM AUTHOR]
- Published
- 2013
- Full Text
- View/download PDF
20. Metabolic engineering for the production of polyunsaturated fatty acids by oleaginous fungus Mortierella alpina 1S-4.
- Author
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Sakuradani, Eiji, Ando, Akinori, Shimizu, Sakayu, and Ogawa, Jun
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BIOENGINEERING , *UNSATURATED fatty acids , *FATTY acid synthesis , *MORTIERELLA , *DIETARY supplements , *FISH oils as feed , *TRIGLYCERIDES - Abstract
Researches related with the application of functional lipids such as polyunsaturated fatty acids (PUFAs) have been conducted in various fields with a view to health and dietary requirements. Novel rich sources other than known natural sources such as plant seeds and fish oils are required for increasing demands of PUFAs. The filamentous fungus Mortierella alpina 1S-4 produces triacylglycerols rich in arachidonic acid, i.e., ones reaching 20 g/l in concentration and containing 30–70% arachidonic acid as total fatty acids. Various mutants derived from M. alpina 1S-4 have led to the production of oils containing various PUFAs. Molecular breeding of M. alpina strains by means of manipulation of the genes involved in PUFA biosynthesis facilitates improvement of PUFA productivity and elucidation of the functions of their enzymes. This review describes practical PUFA production through mutant breeding, functional analyses of the genes of the enzymes involved in PUFA biosynthesis, and recent advances in unique PUFA production through molecular breeding. [Copyright &y& Elsevier]
- Published
- 2013
- Full Text
- View/download PDF
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