To establish in vitro mass production system of grape anthocyanin pigments through callus and cell suspension culture, the effects of nitrogen amount and the ratio of $NO_3^-$/$NH_4^+$ in the medium on cell growth and anthocyanin production were investigated. Total nitrogen amount and the ratio of $NO_3^-$/$NH_4^+$ in the medium strongly affected anthocyanin production and cell growth. When $NH_4^+$ was fixed, the cell growth was promoted by 50 mM total nitrogen (20 mM $NO_3^-$ : 30 mM $NH_4^+$ ) than other nitrogen combinations, and was strongly inhibited when $NO_3^-$ was lacking (0 mM $NO_3^-$ : 60 mM $NH_4^+$ ) while anthocyanin production was increased. When $NO_3^-$ was fixed, the cell growth was promoted by 70 mM total nitrogen (40 mM $NO_3^-$ : 30 mM $NH_4^+$) than other nitrogen combinations, and was strongly inhibited when $NO_3^-$ was lacking (0 mM $NO_3^-$ : 60 mM $NH_4^+$ ) while anthocyanin production was increased. Cell growth was gradually increased by all nitrogen combinations, but anthocyanin production reached its peak on day 4 in culture. Anthocyanin content increased with decreasing cell density. Sucrose was rapidly hydrolyzed to fructose and glucose within 4 days. Glucose and fructose concentrations in the medium increased and peaked at the 4th day. The anthocyanin content of $NH_4^+$-free 2% sucrose media was 2 times (200 $\mu\textrm{g}$/g) higher than that of 1% sucrose. When $NO_3^-$ was lacking, the highest anthocyanin production was observed at 4% sucrose after 12 days of culture, and increased along with the sucrose concentration.
An enzymatic reaction system was developed and optimized for bioconversion of resveratrol from glucose. Liquid enzyme extracts were prepared from Alternaria sp. MG1, an endophytic fungus from grape, and used directly or after immobilization with sodium alginate. When the enzyme solution was used, efficient production of resveratrol was found within 120 min in a manner that was pH-, reaction time-, enzyme amount-, substrate type-, and substrate concentration-dependent. After the optimization experiments using the response surface methodology, the highest value of resveratrol production (224.40 μg/l) was found under the conditions of pH 6.84, 0.35 g/l glucose, 0.02 mg/l coenzyme A, and 0.02 mg/l ATP. Immobilized enzyme extracts could keep high production of resveratrol during recycling use for two to five times. The developed system indicated a potential approach to resveratrol biosynthesis independent of plants and fungal cell growth, and provided a possible way to produce resveratrol within 2 h, the shortest period needed for biosynthesis of resveratrol so far.
Commercial flake graphite cast iron substrate was coated with titanium powder by low pressure plasma spraying and was irradiated with a $CO_2$ laser to produce the wear resistant composite layer. From the experimental results of this study, it was possible to composite TiC particles on the surface layer by direct reaction between carbon existed in the cast iron matrix and titanium with thermal sprayed coating by remelting and alloying them using laser irradiation. The cooling rate of laser remelted cast iron substrate without titanium coating was about $1{\times}10^4$ K/s to $1{\times}10^5$ K/s in the order under the condition used in this study. The microstructure of alloyed layer consisted of three zones, that is, TiC particule crystallized zone (MHV $400{\sim}500$), the mixed zone of TiC particule+ledebulite (MHV $650{\sim}900$) and the ledebulite zone (MHV $500{\sim}700$). TiC particules were crystallized as a typical dendritic morphology. The secondary TiC dendrite arms were grown to the polygonized shape and were necking. And then the separated arms became cubic crystal of TiC at the slowly solidified zone. But in the rapidly solidified zone of fusion boundry, the fine granular TiC particules were grouped like grape.
Objectives: Gallic acid (GA) is the major component of tannin which could be easily founded in various natural materials such as green tea, red tea, grape juice, and Corni Fructus. The purpose of this study is to investigate the effect of Gallic acid (GA) on production of interleukin (IL) in mouse macrophage Raw 264.7 cells stimulated by lipopolysaccharide (LPS). Methods: Productions of interleukins were measured by High-throughput Multiplex Bead based Assay with Bio-plex Suspension Array System based on $xMAP^{(R)}$ (multi-analyte profiling beads) technology. Firstly, cell culture supernatant was obtained after treatment with LPS and GA for 24 hour. Then, it was incubated with the antibody-conjugated beads for 30 minutes. And detection antibody was added and incubated for 30 minutes. And Strepavidin-conjugated Phycoerythrin (SAPE) was added. After incubation for 30 minutes, the level of SAPE fluorescence was analyzed on Bio-plex Suspension Array System and concentration of interleukin was determined. Results: The results of the experiment are as follows. 1. GA significantly inhibited the production of IL-3, IL-10, IL-12p40, and IL-17 in LPS-induced mouse macrophage RAW 264.7 cells at the concentration of 25, 50, 100, 200 uM (p<0.05). 2. GA significantly inhibited the production of IL-6 in LPS-induced mouse macrophage RAW 264.7 cells at the concentration of 50, 100, 200 uM (p<0.05). 3. GA diminished the production of some cytokine such as IL-4, IL-5, and IL-13 in LPS-induced mouse macrophage RAW 264.7 cells. 4. GA did not show the inhibitory effect on the production of IL-$1{\alpha}$ and IL-9 in LPS-induced mouse macrophage RAW 264.7 cells. Conclusions: These results suggest that GA has anti-inflammatory activity related with its inhibitory effects on the production of interleukins such as IL-3, IL-10, IL-12p40, IL-17, and IL-6 in LPS-induced macrophages.
Journal of Physiology & Pathology in Korean Medicine
/
v.24
no.4
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pp.586-591
/
2010
Chemokine and Growth Factor are major mediumtors of immuno-inflammatory pathway. The purpose of this study is to investigate whether productions of Chemokine and Growth Factor in lipopolysaccharide (LPS)-induced mouse macrophage RAW 264.7 cells are modulated by Gallic acid (GA), which is easily founded in tannin-containing natural materials such as red wine, green tea, grape juice, and Corni Fructus. Productions of Chemokine and Growth Factor were analyzed by High-throughput Multiplex Bead based Assay with Bio-plex Suspension Array System based on $xMAP^{(R)}$ (multi-analyte profiling beads) technology. At first, cell culture supernatant was obtained after treatment with LPS and GA for 24 hour. Then, the antibody-conjugated beads were added and incubated for 30 minutes. After incubation, detection antibody was added and incubated for 30 minutes. And Strepavidin-conjugated Phycoerythrin (SAPE) was added. After incubation for 30 minutes, the level of SAPE fluorescence was analyzed on Bio-plex Suspension Array System. Based on fluorescence intensity, concentrations of Chemokine and Growth Factor were determined. The results of the experiment are as follows. GA significantly inhibited the production of interferon-inducible protein (IP)-10, keratinocyte-derived chemokine(KC), and vascular endothelial growth factor (VEGF) in LPS-induced RAW 264.7 cells at the concentration of 25, 50, 100, 200 uM (p<0.05). GA significantly inhibited the production of monocyte chemoattractant protein-1(MCP-1) and macrophage-colony stimulating factor(M-CSF) in LPS-induced RAW 264.7 cells at the concentration of 50, 100, 200 uM (p<0.05). GA diminished the production of granulocyte macrophage-colony stimulating factor (GM-CSF) in LPS-induced RAW 264.7 cells. But GA did not show the inhibitory effect on the production of leukemia inhibitory factor (LIP) and macrophage inflammatory protein (MIP)-2 in LPS-induced RAW 264.7 cells. These results suggest that GA has the immuno-modulating activity related with its inhibitory effects on the production of IP-10, KC, MCP-1, VEGF, and M-CSF in LPS-induced macrophages.
During the production of grape wine, the formation of thick leathery pellicle/bacterial cellulose (BC) at the airliquid interface was due to the bacterium, which was isolated and identified as Gluconacetobacter hansenii UAC09. Cultural conditions for bacterial cellulose production from G. hansenii UAC09 were optimized by central composite rotatable experimental design. To economize the BC production, coffee cherry husk (CCH) extract and corn steep liquor (CSL) were used as less expensive sources of carbon and nitrogen, respectively. CCH and CSL are byproducts from the coffee processing and starch processing industry, respectively. The interactions between pH (4.5-8.5), CSL (2-10%), alcohol (0.5-2%), acetic acid (0.5-2%), and water dilution rate to CCH ratio (1:1 to 1:5) were studied using response surface methodology. The optimum conditions for maximum BC production were pH (6.64), CSL (10%), alcohol (0.5%), acetic acid (1.13%), and water to CCH ratio (1:1). After 2 weeks of fermentation, the amount of BC produced was 6.24 g/l. This yield was comparable to the predicted value of 6.09 g/l. This is the first report on the optimization of the fermentation medium by RSM using CCH extract as the carbon source for BC production by G. hansenii UAC09.
Jeong, Yu Jeong;Woo, Su Gyeong;An, Chul Han;Jeong, Hyung Jae;Hong, Young-Soo;Kim, Young-Min;Ryu, Young Bae;Rho, Mun-Chual;Lee, Woo Song;Kim, Cha Young
Molecules and Cells
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v.38
no.4
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pp.318-326
/
2015
We previously reported that the SbROMT3syn recombinant protein catalyzes the production of the methylated resveratrol derivatives pinostilbene and pterostilbene by methylating substrate resveratrol in recombinant E. coli. To further study the production of stilbene compounds in E. coli by the expression of enzymes involved in stilbene biosynthesis, we isolated three stilbene synthase (STS) genes from rhubarb, peanut, and grape as well as two resveratrol O-methyltransferase (ROMT) genes from grape and sorghum. The ability of RpSTS to produce resveratrol in recombinant E. coli was compared with other AhSTS and VrSTS genes. Out of three STS, only AhSTS was able to produce resveratrol from p-coumaric acid. Thus, to improve the solubility of RpSTS, VrROMT, and SbROMT3 in E. coli, we synthesized the RpSTS, VrROMT and SbROMT3 genes following codon-optimization and expressed one or both genes together with the cinnamate/4-coumarate:coenzyme A ligase (CCL) gene from Streptomyces coelicolor. Our HPLC and LC-MS analyses showed that recombinant E. coli expressing both ScCCL and RpSTSsyn led to the production of resveratrol when p-coumaric acid was used as the precursor. In addition, incorporation of SbROMT3syn in recombinant E. coli cells produced resveratrol and its mono-methylated derivative, pinostilbene, as the major products from p-coumaric acid. However, very small amounts of pterostilbene were only detectable in the recombinant E. coli cells expressing the ScCCL, RpSTSsyn and SbROMT3syn genes. These results suggest that RpSTSsyn exhibits an enhanced enzyme activity to produce resveratrol and SbROMT3syn catalyzes the methylation of resveratrol to produce pinostilbene in E. coli cells.
Journal of the Korea Academia-Industrial cooperation Society
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v.18
no.5
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pp.581-586
/
2017
The facility modernization policy has been established to improve fruits quality and to increase fruits yield per acreage. The fruit production quantity of farms joined in the policy was increased. Therefore, many fruit farms want to participate in the policy. The government has subsidized fruit farms to modernize their facilities such as rain proof, drainage way, frost proof, etc. This study analyzes the performance of the facility modernization policy focused on apple, eastern pear, and grape cultivation sector. One hundred apple farms, one hundred eastern pear farms, and 91 grape farms were surveyed. The performance of the policy was reviewed using analytical technique such as Covariate Matching and Propensity Score Matching and several policy implications were suggested.
Danored is considered as the most favorable grape variety for national wine manufacturing in terms of sugar, organic acid contents, annual production amount, as well as quality control convenience after harvest. This study was canied out to know the influence of suspended solid(SS) and skin contact time(SCT) with Danored variety on the white wine quality. The results are summarized as follows; 1. Suspended solid and skin contact time did not greatly influence on the chemical composition of Danored juice. But skin contact time provided slightly increaseness to the content of phenol compounds 2. Suspended solid accelerated fermentation rate of white wine manufacturing. Although the content of suspended solid are less amount than 0.3% in must, wine fermentation was completed to dryness in concentration of sugar. 3. When the content of suspended solid was around 3%, higher alcohol in white wine was increased by the 20%. This result is relatively small increasement compared to the other reported results. 4. Because Danored contained less amount of phenol compounds comparison with other grape varieties, browning capacity in white wine was much favorable to quite low concentration. Especially, phenolic compounds was decreased remarkably as one of fermentation characteristics of Danored variety. 5. As a result of sensory evaluation, wine quality was A-1> B-1> A-2> B-2 in its favorable ranking orders. It was so concluded that A-1 might be the most acceptable one from this study.
Irrigation applications (IA) and increased bud load (BL) are fundamental practices that are used to achieve optimum yields in grape production, while maintaining fruit quality parameters. Two different irrigation amounts (IA-I and IA-II) based on growth stages, in addition to a non-irrigated (rain-fed) control, along with two different BL applications [normal bud load based on traditional pruning practices (1BL) and double bud load (2BL)] were evaluated over a two-year experiment for their effects on the biochemical composition of the berries. Berries from the rain-fed vines had higher sugar levels, whereas no significant change was detected in organic acid levels. The increased bud load (2BL) treatment yielded less sugar in the berries compared to the 1BL control in both years. The total non-colored phenolic compounds (NPC) were greater in the irrigated (especially IA-I) and 2BL treatments than in those of the rain-fed and 1BL control. However, total anthocyanin was greater in the non-irrigated and 1BL control than in the irrigated and 2BL treatments. The antioxidant contents of the berries also varied according to the treatments and years. Our results implied that implementing a higher bud load along with the IA-I irrigation application, in which irrigation applications were 50 and 75% of the cumulative evaporation from the Class A pan during berry set to veraison and veraison to harvest growth stages, respectively, can help in obtaining greater yields in high-plateau viticulture. Thus, if more buds are left on the vines, along with sufficient irrigation and rainfall, yield may increase while maintaining or increasing the biochemical composition of the berries.
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