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Sphingorhabdus sp. YGSMI21, a novel microbial strain with an enantioselective epoxide hydrolase activity, was isolated from tidal samples contaminated by accidental oil spills subjected to enriched culture with polycyclic aromatic hydrocarbon. This strain was able to optically decompose (R)-styrene oxide (SO) and showed 100% optical purity. In addition, it showed a good enantioselectivity for the derivatives of (S)-SO, (S)-2-chlorostyrene oxide (CSO), (S)-3-CSO and (S)-4-CSO. For (S)-2-CSO, (S)-3-CSO and (S)-4-CSO, 99.9%ee was obtained with the yield of 26.2%, 24.8%, and 11.0%, respectively, when using 10 mg cells of Sphingorhabdus sp. YGSMI21 at pH 8.0 with 4 mM racemic substrates at pH 8.0 and 25°C. The values obtained in this study for (S)-2-CSO, particularly the yield of 26.2%, is noteworthy, considering that obtaining an enantiomerically pure form is difficult. Taken together, Sphingorhabdus sp. YGSMI21 can be regarded as a whole-cell biocatalyst in the production of various (S)-CSO with the chlorine group at a different position.

Background and purpose Sea weeds have been used since ancient times in Asian countries, especially in Korea, Japan, and China, as both edible sea vegetables and traditional medicinal tonics due to their health benefits. Eisenia bicyclis has been studied for anti-allergic and anti-cancer effects; however, its effects on the cardiovascular system, especially on platelet function, are yet to be explored. Therefore, we examined the effect of E. bicyclis on platelet function. Study design and methods E. bicyclis extract (EBE) was prepared and in vitro effects on ADP-induced platelet aggregation, granule secretion, intracellular calcium ion ([Ca2+]i) mobilization, fibrinogen binding to integrin αIIbβ3 and clot retraction were evaluated. Phosphorylation levels of MAPK signaling molecules and P2Y12 receptor downstream signaling pathway components were studied. In vivo effects were studied using an arteriovenous (AV) shunt model. Results EBE markedly inhibited in vitro ADP-induced platelet aggregation, granule secretion (ATP release and P-selectin expression), [Ca2+]i mobilization, fibrinogen binding to integrin αIIbβ3, and clot retraction; attenuated MAPK pathway activation; and inhibited phosphorylation of PI3K/Akt, PLCγ2, and Src. The extract significantly inhibited in vivo thrombus weight in an AV shunt model. Conclusion E. bicyclis inhibits agonist-induced platelet activation and thrombus formation through modulation of the P2Y12 receptor downstream signaling pathway, suggesting its therapeutic potential in ethnomedicinal applications as an anti-platelet and anti-thrombotic compound to prevent cardiovascular diseases.

This study aimed to investigate the various biological activities of Geranium thunbergii such as antimicrobial activity and protective effect against oxidative damage. To evaluate its antioxidant and antimicrobial activities, we first performed methanol extraction; this methanol extract was further partitioned using various solvents. And then, its antioxidant activity was measured using various assays including total phenolic content and protection against oxidative DNA damage, and antimicrobial activities were examined using minimum inhibiting concentration (MIC) test, and paper disc method. In addition, high-performance liquid chromatography was performed to analyze the major chemical components of ethyl acetate fraction. The G. thunbergii fraction with ethyl acetate exhibited higher antioxidant and antimicrobial activities than the other fractions. The results showed that G. thunbergii ethyl acetate fraction at 50 μg/mL had strong DPPH and ABTS radical scavenging activities of 80.88% and 80.12%, respectively. In addition, the ethyl acetate fraction protected DNA from the oxidative damage induced by ferrous ion and hydroxyl radicals and showed high antimicrobial activity with diameter of inhibition zones ranging from 13.33 to 15.67 mm. High-performance liquid chromatography analysis revealed the major phenolic compounds of G. thunbergii to be ellagic acid and gallic acid. These results suggest that G. thunbergii might protect DNA against oxidative stress induced by reactive oxygen species and can be utilized as a natural source of antioxidant and antimicrobial agent in the food industry.

Geranium thunbergii is a perennial plant commonly used as an oriental medicine for prevention of diarrhea, constipation, and gastrointestinal disorders. However, its anti-inflammatory effect has not been evaluated thus far. Therefore, the present study aimed to investigate the anti-inflammatory effect of G. thunbergii. In this study, G. thunbergii extracted with methanol; this methanol extract was further partitioned using various solvents, and G. thunbergii ethyl acetate fraction (GTEF) was obtained. To determine the anti-inflammatory activity of G. thunbergii, the effects of GTEF on nitric oxide (NO) production in lipopolysaccharide-stimulated RAW 264.7 cells were evaluated. GTEF suppressed NO production in a dose-dependent manner without any toxic effects. In addition, western blotting was performed to examine the effect of GTEF on expression of inducible nitric oxide synthase and cyclooxygenase-2. These results suggest that GTEF as a phytoextract may be useful for the prevention or treatment of inflammation.

Sphingorhabdus sp. YGSMI21 is a novel strain exhibiting high enantioselective hydrolysis activity for styrene oxide. Here, we present its complete genome sequence, consisting of one circular chromosome (3.86 Mb) and one plasmid (0.196 Mb).

Marine bacteria possessing epoxide hydrolase (EH) activity were isolated from the oil-spilled foreshore of South Korea. While the isolated Rhodococcus sp. YSMI04 and YSNA32 showed different enantioselectivities to racemic styrene oxide (SO), two strains exhibited same enantiopreference to 3-chlorostyrene oxide (3-CSO) and 4-CSO. In the case of 3-CSO, Rhodococcus sp. YSMI04 and YSNA32 exhibited (R)-3-CSO preferred hydrolysis activity, whereas Roseobacter sp. TSBP12 showed a preference for (S)-3-CSO. In the case of 4-CSO, Rhodococcus sp. YSMI04 and YSNA32 showed enantiopreference to (S)-4-CSO, Roseobacter sp. TSBP12 exhibited (R)-4-CSO preferred activity. (S)-4-CSO was obtained with 35% yield (theoretically 50%) using Roseobacter sp. TSBP12.

Sea weeds have been used since ancient times in Asian countries, especially in Korea, Japan, and China, as both edible sea vegetables and traditional medicinal tonics due to their health benefits. Eisenia bicyclis has been studied for anti-allergic and anti-cancer effects; however, its effects on the cardiovascular system, especially on platelet function, are yet to be explored. Therefore, we examined the effect of E. bicyclis on platelet function.E. bicyclis extract (EBE) was prepared and in vitro effects on ADP-induced platelet aggregation, granule secretion, intracellular calcium ion ([CaEBE markedly inhibited in vitro ADP-induced platelet aggregation, granule secretion (ATP release and P-selectin expression), [CaE. bicyclis inhibits agonist-induced platelet activation and thrombus formation through modulation of the P

BACKGROUND Brown algae have been used for their nutritional value as well as a source of bioactive compounds with antioxidant, anti-inflammatory, antimicrobial and anti-obesity effects. Obesity is an important condition implicated in various diseases, including diabetes, hypertension, dyslipidemia and coronary heart disease. However, anti-obesity effects of Eisenia bicyclis remain unknown. RESULTS We investigated the anti-obesity effects of 6,6′-bieckol, 6,8′-bieckol, 8,8′-bieckol, dieckol and phlorofucofuroeckol A isolated from E. bicyclis. Anti-obesity activity was evaluated by examining the inhibition of differentiation of 3T3-L1 adipocytes and the expression of peroxisome proliferator-activated receptor γ (PPARγ), CCATT/enhancer-binding protein α (C/EBPα) and sterol regulatory element binding protein-1c (SREBP-1c) at the mRNA and protein level. Differentiated 3T3-L1 cells were treated with the purified phlorotannins at concentrations of 10, 25 and 50 µg mL−1 for 8 days. The results indicated that the purified phlorotannins suppressed the differentiation of 3T3-L1 adipocytes in a dose–dependent manner, without toxic effects. Among the five compounds, 6,6′-bieckol markedly decreased lipid accumulation and expression levels of PPARγ, C/EBPα, SREBP-1c (mRNA and protein), and fatty acid synthase and acyl-coA carboxylase (mRNA). CONCLUSION These findings suggest that E. bicyclis suppressed differentiation of 3T3-L1 adipocyte through downregulation of adipogenesis and lipogenesis. © 2014 Society of Chemical Industry

In this study, we determined 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical scavenging activities as well as the reducing power for screening of antioxidant activity of bieckols in Eisenia bicyclis (Kjellamn) Setchell (E. bicyclis). The compounds 6,6′-bieckol, 6,8′-bieckol and 8,8′-bieckol are representative members of the phlorotannins family. The isolated bieckols displayed markedly strong DPPH and ABTS radical scavenging effects when compared to those of positive controls (BHA and Trolox). These bieckols were also found to have significant reducing power. The isolated bieckols (6,6′-bieckol, 6,8′-bieckol and 8,8′-bieckol) were found to effectively suppress the detrimental effects of 1O2 on type-II photosensitization. The concentrations required to exert a 50 % quenching effect on 1O2 (QC50) were found to be 30.7, 35.7 and 49.4 μM for 6,6′-bieckol, 6,8′-bieckol and 8,8′-bieckol, respectively. Interestingly, all these bieckols were found to be superior to histidine (5.9 mM), a well-known 1O2 quencher. These results suggested that brown algae phlorotannins, and bieckols in particular, may play an important role in protecting marine organisms against sunlight damage by eliminating 1O2. This knowledge may contribute to the development of natural bioactive products with potential applications in reducing photo-produced oxidative damage involving reactive oxygen species in living organisms.

In this study, we aimed to elucidate the antioxidant capacity of Eisenia bicyclis and evaluated its antioxidant activity using various assay systems such as 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, 2,2-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical scavenging, reducing power ability, and content of total polyphenol. Among all the performed experiments, the ethyl acetate fraction of E. bicyclis exhibited higher antioxidant activities. From this finding, isolation and purification were performed on the ethyl acetate fraction and identified dieckol and phlorofucofureoeckol-A by spectroscopic analyses including FAB-mass in the negative mode, (1) H NMR, (13) C NMR, (1) H-(1) H COSY, HMQC, and HMBC spectra. Interestingly, ABTS radical scavenging activities of dieckol and phlorofucofuroeckol showed strong effects of 65.36% and 70.38% at a concentration of 50 μg/mL, respectively. DPPH radical scavenging and reducing power abilities were increased in a dose-dependent manner at various concentrations. These results suggest that dieckol and phlorofucofuroeckol-A of E. bicyclis may play an important role in protection from oxidative stress involving reactive oxygen species and may contribute to the development of new bio products, for example, a useful preservative to improve food quality and a drug for various oxidative damage-associated diseases. Practical Application: The results suggest that dieckol and phlorofucofuroeckol-A can be utilized as a natural source for potential application of antioxidant in food industry and drug for oxidative damage-associated diseases.

A β-galactosidase from Thermotoga maritima produced galacto-oligosaccharides (GOS) from lactose by transgalactosylation when expressed in Escherichia coli. The enzyme activity for GOS production was maximal at pH 6.0 and 90 °C. In thermal stability experiments, the enzyme followed first-order kinetics of pH and thermal inactivation, and half-lives at pH 5.0, pH 8.0, 80 °C, and 95 °C were 27 h, 82 h, 41 h, and 14 min, respectively, suggesting that the enzyme was stable below 80 °C and in the pH range of 5.0–8.0. Mn2+ was the most effective divalent cation for GOS production. Cu2+ and EDTA inhibited more than 84% of enzyme activity. GOS production increased with increasing lactose concentrations and peaked at 500 g lactose/l. Among tested enzyme concentrations, the highest production of GOS was obtained at 1.5 units enzyme/ml. Under the optimal conditions of pH 6.0, 80 °C, 500 g lactose/l, and 1.5 units enzyme/ml, GOS production was 91 g/l for 300 min, with a GOS productivity of 18.2 g/l · h and a conversion yield of GOS to lactose of 18%.

We found that certain Str-, Gen- or Rif- mutants derived from Pseudomonas putida KH146-2, which are resistant to streptomycin, gentamicin or rifampicin, respectively, are tolerant to the aromatic compound 4-hydroxybenzoate (4HBA). The minimum inhibitory concentration (MIC) of 4HBA as the sole carbon source for the wild-type strain was 1%, whereas the MIC for the mutants was 1.7%. Frequency of 4HBA-tolerant mutants among spontaneous Str-, Gen- and Rif- mutants was 5-15%, 3-5%, and 3% respectively. These 4HBA-tolerant mutants also tolerated to a variety of organic chemicals such as 3-hydroxybenzoate, aliphatic and heterocyclic compounds, chlorobenzoates, as well as organic solvents toluene and m-xylene. The Str mutants had a point mutation in the rpsL gene, which produces the ribosomal protein S12. The Rif mutants were found to have a point mutation in the rpoB gene, which encodes the RNA polymerase beta-subunit. Mutation points in Gen mutants still remain unknown. Str-, Gen- and Rif-phenotypes occurred in spontaneous 4HBA-tolerant mutants which had been selected by successively increasing concentrations (from 0.8% to 5%) of 4HBA. Complementation experiments with one of the Str mutants demonstrated a causal relationship between a rpsL mutation (str-1) and 4HBA tolerance. Uptake experiments using [14C]-4HBA revealed that apparent ability of 4HBA to be taken up by the membrane transport system was reduced two to threefold in the mutants compared to the wild-type strain, accounting at least partly for the enhanced tolerance to 4HBA. Our approaches thus could be effective in improvement of tolerance to aromatic compounds of bacteria applicable for bioremediation.

Limited proteolysis of papain on the intact family F/10 β-xylanase (45 kDa) led to the isolation of catalytic domain (32 kDa) and xylan-binding domain (15 kDa). The two truncated protein fragments were purified by gel filtration to homogeneity. The purified catalytic domain was fully active against 4-O-methyl-d-glucurono-d-xylan, and the action mode on xylan was the same as the intact xylanase. However, the removal of xylan-binding domain reduces dramatically the capability of adsorption for xylan.

Eisenia bicyclis is edible brown algae recognized as a rich source of bioactive derivatives mainly phlorotannins reported for their anti-oxidant properties. Of all phlorotannins identified so far, dieckol has shown the most potent effect in anti-inflammatory, radical scavenging and neuroprotective functions. However, whether dieckol up-regulates hemeoxygenase 1 (HO-1) and this mediates its anti-inflammatory effect in murine macrophages remains poorly understood. Dieckol (12.5-50 μM) inhibited nitric oxide production and attenuated inducible nitric oxide synthase, phospho (p)-PI-3K, p-Akt, p-IKK-α/β, p-IκB-α and nuclear p-NF-κBp65 protein expressions, and NF-κB transcriptional activity in LPS (0.1 μg/ml) stimulated murine macrophages. On the other hand, dieckol up-regulated HO-1 which partly mediated its anti-inflammatory effect in murine macrophages. Thus, dieckol appeared to be a potential therapeutic agent against inflammation through HO-1 up-regulation.

Brown algae have been used for their nutritional value as well as a source of bioactive compounds with antioxidant, anti-inflammatory, antimicrobial and anti-obesity effects. Obesity is an important condition implicated in various diseases, including diabetes, hypertension, dyslipidemia and coronary heart disease. However, anti-obesity effects of Eisenia bicyclis remain unknown.We investigated the anti-obesity effects of 6,6'-bieckol, 6,8'-bieckol, 8,8'-bieckol, dieckol and phlorofucofuroeckol A isolated from E. bicyclis. Anti-obesity activity was evaluated by examining the inhibition of differentiation of 3T3-L1 adipocytes and the expression of peroxisome proliferator-activated receptor γ (PPARγ), CCATT/enhancer-binding protein α (C/EBPα) and sterol regulatory element binding protein-1c (SREBP-1c) at the mRNA and protein level. Differentiated 3T3-L1 cells were treated with the purified phlorotannins at concentrations of 10, 25 and 50 µg mL(-1) for 8 days. The results indicated that the purified phlorotannins suppressed the differentiation of 3T3-L1 adipocytes in a dose-dependent manner, without toxic effects. Among the five compounds, 6,6'-bieckol markedly decreased lipid accumulation and expression levels of PPARγ, C/EBPα, SREBP-1c (mRNA and protein), and fatty acid synthase and acyl-coA carboxylase (mRNA).These findings suggest that E. bicyclis suppressed differentiation of 3T3-L1 adipocyte through downregulation of adipogenesis and lipogenesis.

Twenty-three kinds of commercially available enzymes were subjected to the screening test for xylanolytic activities, namely, endo-β-xylanase, β-xylosidase, α-arabinofuranosidase and α-glucuronidase.The majority of the enzymes contained some of the activities. The above enzymes were also submitted to the hydrolyzable test on the arabinoglucuronoxylan of corn hull. Among them, Cellulase C-0901 (Sigma Chemicals) originating from Penicillium funiculosum, although lacking in a-glucuronidase, degraded the xylan very well, and released 80.5% of arabinose and 60.8% of xylose based on the component sugars of the xylan. On the other hand, Cellulase 'Onozuka' R-10 (Yakult Co.) originating from Trichoderma viride, although containing all of the xylanolytic activities, decomposed hardly any of the arabinoglucuronoxylan. According to these results, commer cially available enzymes contained high and various xylanolytic activities. However, the intensities of enzyme activity and hydrolyzable ability differed remarkably in the degradation of arabinogluc uronoxylan. In addition, Cellulase C-0901 was most suitable among them for the enzymatic hydrolysis of the xylan.

Arabinoxylan (`Cellace') corn fiber, containing 28.1% (w/w) as l-arabinose and 32.8% (w/w) as d-xylose, was hydrolyzed by a crude enzyme containing β-xylanase, β-xylosidase and α-l-arabinofuranosidase originating from the extracellular culture broth of Penicillium funiculosum. The resultant hydrolysate contained l-arabinose, d-xylose and small amounts of other mono- and oligosaccharides. The l-arabinose and d-xylose were 21.3% (w/w) and 18.7% (w/w), respectively, based on the initial arabinoxylan. Williopsis saturnus var. saturnus, which metabolizes d-xylose without using l-arabinose, was aerobically cultured in the hydrolysate at 30 °C for 96 h. The sugar solution after removal of yeast cells contained l-arabinose and d-xylose which were 20.3% (w/w) and 0.002% (w/w), respectively, of the initial arabinoxylan. The solution was decolorized with activated carbon, and deionized with cation- and anion-exchange resins. The clear sugar solution thus obtained was composed of l-arabinose and d-xylose which were 19.3% (w/w) and 0.001% (w/w), respectively, of the initial arabinoxylan. After concentration of the sugar solution under reduced pressure, followed by crystallization of l-arabinose, 16% (w/w) l-arabinose (based on the initial arabinoxylan) was obtained as crude crystals. No d-xylose was detected in the final preparation.

Production of the artificial sweetener, lactosucrose, by various microorganisms containing levansucrase activity was investigated. Of the tested bacteria, Bacillus subtilis was the most effective producer using lactose as an acceptor and sucrose as a fructosyl donor. Lactosucrose production by this strain was optimal at pH 6.0 and 55 degrees C whereupon 181 g lactosucrose l(-1) was produced from 225 g lactose l(-1) and 225 g sucrose l(-1) in 10 h.

In this study, we aimed to elucidate the antioxidant capacity of Eisenia bicyclis and evaluated its antioxidant activity using various assay systems such as 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, 2,2-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical scavenging, reducing power ability, and content of total polyphenol. Among all the performed experiments, the ethyl acetate fraction of E. bicyclis exhibited higher antioxidant activities. From this finding, isolation and purification were performed on the ethyl acetate fraction and identified dieckol and phlorofucofureoeckol-A by spectroscopic analyses including FAB-mass in the negative mode, (1) H NMR, (13) C NMR, (1) H-(1) H COSY, HMQC, and HMBC spectra. Interestingly, ABTS radical scavenging activities of dieckol and phlorofucofuroeckol showed strong effects of 65.36% and 70.38% at a concentration of 50 μg/mL, respectively. DPPH radical scavenging and reducing power abilities were increased in a dose-dependent manner at various concentrations. These results suggest that dieckol and phlorofucofuroeckol-A of E. bicyclis may play an important role in protection from oxidative stress involving reactive oxygen species and may contribute to the development of new bio products, for example, a useful preservative to improve food quality and a drug for various oxidative damage-associated diseases. Practical Application: The results suggest that dieckol and phlorofucofuroeckol-A can be utilized as a natural source for potential application of antioxidant in food industry and drug for oxidative damage-associated diseases.

We found that certain Str-, Gen- or Rif- mutants derived from Pseudomonas putida KH146-2, which are resistant to streptomycin, gentamicin or rifampicin, respectively, are tolerant to the aromatic compound 4-hydroxybenzoate (4HBA). The minimum inhibitory concentration (MIC) of 4HBA as the sole carbon source for the wild-type strain was 1%, whereas the MIC for the mutants was 1.7%. Frequency of 4HBA-tolerant mutants among spontaneous Str-, Gen- and Rif- mutants was 5-15%, 3-5%, and 3% respectively. These 4HBA-tolerant mutants also tolerated to a variety of organic chemicals such as 3-hydroxybenzoate, aliphatic and heterocyclic compounds, chlorobenzoates, as well as organic solvents toluene and m-xylene. The Str mutants had a point mutation in the rpsL gene, which produces the ribosomal protein S12. The Rif mutants were found to have a point mutation in the rpoB gene, which encodes the RNA polymerase beta-subunit. Mutation points in Gen mutants still remain unknown. Str-, Gen- and Rif-phenotypes occurred in spontaneous 4HBA-tolerant mutants which had been selected by successively increasing concentrations (from 0.8% to 5%) of 4HBA. Complementation experiments with one of the Str mutants demonstrated a causal relationship between a rpsL mutation (str-1) and 4HBA tolerance. Uptake experiments using [14C]-4HBA revealed that apparent ability of 4HBA to be taken up by the membrane transport system was reduced two to threefold in the mutants compared to the wild-type strain, accounting at least partly for the enhanced tolerance to 4HBA. Our approaches thus could be effective in improvement of tolerance to aromatic compounds of bacteria applicable for bioremediation.

Cotton-seed residual cake, which is a byproduct of the process of oil extraction from the seed, was delignified with sodium hypochlorite (1% available chlorine). Xylan was then prepared from the delignified wet material by alkali extraction with 15% sodium hydroxide. The cotton-seed xylan contained 64.7% xylose and 9.4% uronic acid. The xylan was hydrolyzed with 0.125 M sulfuric acid at 90 degrees C for 15 min. The resultant hydrolysis products were separated by gel-permeation chromatography on BioGel P-4 and Toyopearl HW-40F columns connected in series, with water as an eluate. Xylose and xylooligosaccharides with a degree of polymerization ranging from DP 2 to 15 were separated under such conditions, and each xylooligosaccharide-containing peak fraction afforded a single band on fluorophore-assisted carbohydrate electrophoresis. These results suggest that cotton-seed xylan is suitable for the preparation of xylose and xylooligosaccharides.