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Rare ginsenoside Ia synthesized from F1 by cloning and overexpression of the UDP-glycosyltransferase gene from Bacillus subtilis: synthesis, characterization, and in vitro melanogenesis inhibition activity in BL6B16 cells

  • Wang, Dan-Dan;Jin, Yan;Wang, Chao;Kim, Yeon-Ju;Perez, Zuly Elizabeth Jimenez;Baek, Nam In;Mathiyalagan, Ramya;Markus, Josua;Yang, Deok-Chun
    • Journal of Ginseng Research
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    • v.42 no.1
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    • pp.42-49
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    • 2018
  • Background: Ginsenoside F1 has been described to possess skin-whitening effects on humans. We aimed to synthesize a new ginsenoside derivative from F1 and investigate its cytotoxicity and melanogenesis inhibitory activity in B16BL6 cells using recombinant glycosyltransferase enzyme. Glycosylation has the advantage of synthesizing rare chemical compounds from common compounds with great ease. Methods: UDP-glycosyltransferase (BSGT1) gene from Bacillus subtilis was selected for cloning. The recombinant glycosyltransferase enzyme was purified, characterized, and utilized to enzymatically transform F1 into its derivative. The new product was characterized by NMR techniques and evaluated by MTT, melanin count, and tyrosinase inhibition assay. Results: The new derivative was identified as (20S)-$3{\beta},6{\alpha},12{\beta}$,20-tetrahydroxydammar-24-ene-20-O-${\beta}$-D-glucopyranosyl-3-O-${\beta}$-D-glucopyranoside(ginsenoside Ia), which possesses an additional glucose linked into the C-3 position of substrate F1. Ia had been previously reported; however, no in vitro biological activity was further examined. This study focused on the mass production of arduous ginsenoside Ia from accessible F1 and its inhibitory effect of melanogenesis in B16BL6 cells. Ia showed greater inhibition of melanin and tyrosinase at $100{\mu}mol/L$ than F1 and arbutin. These results suggested that Ia decreased cellular melanin synthesis in B16BL6 cells through downregulation of tyrosinase activity. Conclusion: To our knowledge, this is the first study to report on the mass production of rare ginsenoside Ia from F1 using recombinant UDP-glycosyltransferase isolated from B. subtillis and its superior melanogenesis inhibitory activity in B16BL6 cells as compared to its precursor. In brief, ginsenoside Ia can be applied for further study in cosmetics.

The Effect of Spray Dried Plasma, Lactose and Soybean Protein Sources on the Performance of Weaned Pigs

  • Liu, H.;Kim, I.B.;Touchette, K.J.;Newcomb, M.D.;Allee, G.L.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.9
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    • pp.1290-1298
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    • 2001
  • A total of 371 weaned pigs were used in three experiments to evaluate the effects of spray dried plasma (SDP), soybean protein sources, and lactose on growth performance. In Exp. 1, 128 pigs (5.99 kg, $18{\pm}2d$) were used to evaluate the effect of SDP (0 vs 7%), lactose (0 vs 30%), and two soybean protein sources [soybean meal (SBM) and extruded soybean protein concentrate (ESPC)] in phase I (d 0 to 14) diets on pig performance in a $2{\times}2{\times}2$ factorial arrangement. Spray-dried plasma increased phase I ADG (p<0.01) and ADFI (p<0.05) in the SBM diets, but not in the ESPC diets. Lactose improved ADG and gain/feed ratio (G/F) in phase I (p<0.01). In Exp. 2, 144 pigs (5.50 kg, $17{\pm}3d$) were used to evaluate the effect of SDP (0 vs. 3.5%) and three soybean protein sources [SBM, ESPC, and soybean protein concentrate (SPC)] in phase I diets, and the effects of two different phase II (d 14 to 28) diets (simple vs complex) in a $2{\times}3{\times}2$ factorial arrangement of treatments. In phase I, SDP increased ADG (p<0.01) and improved G/F (p<0.05). Pigs fed SBM had the highest ADG and ADFI, with a G/F similar to the pigs fed ESPC. In phase II, pigs fed the complex diet had improved ADG (p<0.01), ADFI (p<0.05), and G/F (p<0.05) compared to the simple diet. In Exp. 3, 99 weaned pigs (5.77 kg, $17{\pm}3$d) were used to evaluate the effect of SBM, ESPC, and ESPC with SDP in the phase I diets. Pigs fed SBM with no blood product in the diet had the lowest ADG (p<0.01), ADFI (p<0.01), and G/F (p<0.05) in the first week of phase I. There were no differences in soybean protein sources fed in phase I diets on overall pig performance. These experiments are indicated that SDP and lactose improve the phase I performance. Soybean meal can be used as the major protein source in phase I diets with SDP.

Effect of Wood Vinegar on the Performance, Nutrient Digestibility and Intestinal Microflora in Weanling Pigs

  • Choi, J.Y.;Shinde, P.L.;Kwon, I.K.;Song, Y.H.;Chae, Byung-Jo
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.2
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    • pp.267-274
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    • 2009
  • Two experiments were conducted to investigate the feeding value of wood vinegar in weanling pigs. In Experiment 1, weanling pigs (n = 224; Landrace ${\times}$Yorkshire ${\times}$Duroc, 21${\pm}$3 d-old, initial BW 6.12${\pm}$0.10 kg) were assigned to four dietary treatments. Different levels of wood vinegar were added to the diets as dietary treatments (0, 0.1, 0.2 and 0.3%). Each treatment comprised 4 replicates with 14 piglets in each. Experimental feeding was conducted for 28 d in two phases (phase I, d 0 to 14 and phase II, d 15 to 28). Feeding of wood vinegar linearly (p<0.05) improved the phase I, phase II and overall ADG and increased (linear, p<0.05) the overall and phase II ADFI. Linear improvements in the apparent fecal digestibility of dry matter (p = 0.013), gross energy (p = 0.019) and crude protein (p = 0.033) were observed as the level of wood vinegar was increased in the diet of pigs. Experiment 2 was conducted to compare dietary wood vinegar with commonly used growth promoters, organic acid (mixture of 21% phosphoric acid, 3.25% propionic acid, 2.8% formic acid, 10% calcium formate and 5% calcium propionate) and antibiotic (aparamycin). A total of 288 weanling piglets (Landrace ${\times}$Yorkshire ${\times}$Duroc, 22${\pm}$2 d-old, initial BW 6.62${\pm}$0.31 kg) were assigned to four treatments with four replicates (18 piglets/pen) for 28 days and fed in 2 phases: phase I, d 0 to 14 and phase II, d 15 to 28. The dietary treatments were control (corn-soybean meal basal diet without antibiotics) and diets containing 0.2% antibiotic, 0.2% organic acid and 0.2% wood vinegar. Pigs fed antibiotic showed higher (p<0.001) ADG and better feed efficiency followed by pigs fed wood vinegar and organic acid diets while those fed the control diet had lowest ADG and poorest feed efficiency. The overall and phase I ADFI was highest (p<0.001) in pigs fed wood vinegar and lowest in pigs fed the control diet. Apparent fecal digestibility of dry matter, gross energy and crude protein was significantly higher (p<0.05) in pigs fed the antibiotic diet when compared with pigs fed the control but comparable among pigs fed antibiotic, organic acid and wood vinegar diets. Higher populations of Lactobacillus (p = 0.004) were noted in the ileum of pigs fed the wood vinegar diet, while the population of coliforms in the ileum and cecum was higher (p<0.001) in pigs fed the control diet when compared with pigs fed antibiotic, organic acid or wood vinegar diets. These results indicated that wood vinegar could improve the performance of weanling pigs by improving the nutrient digestibility and reducing harmful intestinal coliforms; moreover performance of pigs fed wood vinegar was superior to those fed organic acid.

Performance and Metabolism of Calves Fed Starter Feed Containing Sugarcane Molasses or Glucose Syrup as a Replacement for Corn

  • Oltramari, C.E.;Napoles, G.G.O.;De Paula, M.R.;Silva, J.T.;Gallo, M.P.C.;Pasetti, M.H.O.;Bittar, C.M.M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.7
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    • pp.971-978
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    • 2016
  • The aim of this study was to evaluate the effect of replacing corn grain for sugar cane molasses (MO) or glucose syrup (GS) in the starter concentrate on performance and metabolism of dairy calves. Thirty-six individually housed Holstein male calves were blocked according to weight and date of birth and assigned to one of the starter feed treatments, during an 8 week study: i) starter containing 65% corn with no MO or GS (0MO); ii) starter containing 60% corn and 5% MO (5MO); iii) starter containing 55% corn and 10% MO (10MO); and iv) starter containing 60% corn and 5% GS (5GS). Animals received 4 L of milk replacer daily (20 crude protein, 16 ether extract, 12.5% solids), divided in two meals (0700 and 1700 h). Starter and water were provided ad libitum. Starter intake and fecal score were monitored daily until animals were eight weeks old. Body weight and measurements (withers height, hip width and heart girth) were measured weekly before the morning feeding. From the second week of age, blood samples were collected weekly, 2 h after the morning feeding, for glucose, ${\beta}$-hydroxybutyrate and lactate determination. Ruminal fluid was collected at 4, 6, and 8 weeks of age using an oro-ruminal probe and a suction pump for determination of pH and short-chain fatty acids (SCFA). At the end of the eighth week, animals were harvested to evaluate development of the proximal digestive tract. The composition of the starter did not affect (p>0.05) concentrate intake, weight gain, fecal score, blood parameters, and rumen development. However, treatment 5MO showed higher (p<0.05) total concentration of SCFAs, acetate and propionate than 0MO, and these treatments did not differ from 10MO and 5GS (p>0.05). Thus, it can be concluded that the replacement of corn by 5% or 10% sugar cane molasses or 5% GS on starter concentrate did not impact performance, however it has some positive effects on rumen fermentation which may be beneficial for calves with a developing rumen.

Antioxidant and ACE Inhibiting Activities of Sugared-Buchu (Allium ampeloprasum L. var. porum J. Gay) Fermented with Lactic Acid Bacteria (부추 당침액의 유산균 발효에 따른 항산화 및 ACE저해활성)

  • Lee, Jung-Bok;Bae, Jung-Shik;Son, Il-Kwon;Jeon, Chun-Pyou;Lee, Eun-Ho;Joo, Woo-Hong;Kwon, Gi-Seok
    • Journal of Life Science
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    • v.24 no.6
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    • pp.671-676
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    • 2014
  • In recent years, the growing interest in the health care benefits of sugared-plant fermented enzymes has led to increased consumption. This study investigated the fermentation of sugared-buchu (Leek:sugar, 1:3) by lactic acid bacteria (Lactobacillus acidophilus AML 0422, Lactobacillus brevis HLJ 59, Lactobacillus helveticus AML0410, Lactobacillus plantarium KCTC 13093) and the antibacterial activity, antioxidant activity, and functionality (e.g., anti-hypertensive activity) of the fermented product. The fermented sugared-buchu showed high antibacterial activity against Staphylococcus aureus KCTC 1916, at 31.43 mm, and its total polyphenols, total flavonoid content, and DPPH scavenging activity were 160.8-178 mg/ml, 100-108 mg/ml, and 51.4-58.1%, respectively. DPPH scavenging activity was to that of vitamin C (50 ppm). ACE inhibitory activity was 50.4-67%, depending on the strain of lactic acid bacteria, and the control of sugared-buchu activity was higher than 32.6%. These results suggest that sugared-buchu fermented with lactic acid bacteria has strong antibacterial, antioxidant, and ACE inhibitory activities.

Qualitative PCR Detection of Stack Gene GM Rice (LS28 X Cry1Ac) Developed in Korea (국내개발 stack gene GM 벼(LS28 X Cry1Ac)에 대한 정성 PCR 분석)

  • Shin, Kong-Sik;Park, Jong-Hyun;Lee, Jin-Hyoung;Lee, Si-Myung;Woo, Hee-Jong;Lim, Sun-Hyung;Kim, Hae-Yeong;Suh, Seok-Cheol;Kweon, Soon-Jong
    • Journal of Applied Biological Chemistry
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    • v.52 no.1
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    • pp.1-7
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    • 2009
  • For the development of qualitative PCR detection method of genetically modified (CM) rice, rice species-specific gene, OsCc-1 (rice cytochrome c gene), was selected as suitable far use as an endogenous gene in rice. The primer pair OsCytC-5'/3'with 111 bp amplicon was used for PCR amplification of the rice endogenous gene, OsCc-1 and no amplified product was observed from 8 different crops as templates. Qualitative PCR method was carried out with stack traits of L528$\times$CryIAc1 GM rice developed in Korea. For the qualitative PCRs, some primer pairs were designed with a construct-specific and event-specific type based on T-DNA and junction sequences of T-DNA in GM rice. Actck-5'/3' amplifying between actin promoter and OsCK1 gene introduced in LS28 gave rise to an amplicon 306 bp; also, CrLB-5'/3' from CryIAcl and CKRB-5'/3'amplifying the junction region of T-DNA and genome sequence from LS28 as event-specific primers gave rise to an amplicon 142 bp and 91 bp, respectively. These primer pairs for the detection of event-specific targets not produced PCR amplicons on non-CM rice and various crops in contrast to event lines. Therefore, in this study we verified that event-specific primers were effective to specifically detect stack trait lines and demonstrated that this method presented could be provided with the detection-method data for risk assessment analysis of GM rice to be commercialized.

Effects of the Brown Seaweed Residues Supplementation on In Vitro Fermentation and Milk Production and Composition of Lactating Dairy Cows (미역부산물 첨가가 In Vitro 발효성상과 젖소의 산유량 및 유성분에 미치는 영향)

  • Baek, I.K.;Maeng, W.J.;Lee, S.H.;Lee, H.G.;Lee, S.R.;Ha, J.K.;Lee, S.S.;Hwang, J.H.
    • Journal of Animal Science and Technology
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    • v.46 no.3
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    • pp.373-386
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    • 2004
  • This study was conducted to investigate effects of the brown seaweed residues supplementation on in vitro fermentation, and milk yield and milk composition of dairy cows. Therefore, two experiments consisting of an in vitro and an in vivo growth trial were used. In in vitro experiment, brown seaweed residues(BSR) was supplemented in basal diet with 0, 1, 2 and 4% respectively, and incubated for 3, 6, 9, 12, and 24 h. The pH value, ammonia-N and VFA were investigated. The pH value tended to increase with increasing BSR during the incubation. Particularly, pH was significantly higher in BSR treatments compared with control at 9 h(p < 0.05). While, ammonia-N concentration was not significantly different across treatments during the whole incubation. BSR supplementation did not affect total VFA production, but acetate was linearly increased in BSR treatments compared with control at 12 h(p < 0.05), and its concentration was highest(92.70 mM) in 4% BSR among treatments. The concentration of iso-butyrate tended to increase in BSR treatments in comparison to control during the incubation. In addition, the concentration of iso-valerate was higher in BSR treatments compared with control at 12 and 24 h. In growth trial, BSR was added(800 g/d/animaI) to diets of dairy cow. Dry matter intake was not affected by BSR supplementation, but daily milk yield(kg) significantly increased in BSR treatment compared with control(p < 0.05). However, milk composition(%) and milk yield(kg) were not significantly different between treatments. Milk fat(% and kg/d) tended to slightly decrease in BSR treatment compared with control(3.59% and 1.06 kg/d vs. 3.32% and 1.01 kg/d), The contents of C16:0 and C20:4 in milk significantly increased in BSR treatment compared with control reflecting from dietary fatty acid composition. The content of C18:0 in milk which is end product of biohydrogenation of CI8 unsaturated fatty acids in the rumen significantly increased in BSR treatment compared with control(p < 0.05). C18:2 content in milk tended to decrease, but tended to increase trans-II C18:l and CLA contents in milk in BSR treatment compared with control. In conclusion, it could be summarized that BSR may stabilize rumen pH, and it could improve milk yield and CIA content in milk with more than 4% of diet. Therefore, BSR could be beneficially used in dairy diets as a feed additive.

Two new triterpenoid saponins derived from the leaves of Panax ginseng and their antiinflammatory activity

  • Li, Fu;Cao, Yufeng;Luo, Yanyan;Liu, Tingwu;Yan, Guilong;Chen, Liang;Ji, Lilian;Wang, Lun;Chen, Bin;Yaseen, Aftab;Khan, Ashfaq A.;Zhang, Guolin;Jiang, Yunyao;Liu, Jianxun;Wang, Gongcheng;Wang, Ming-Kui;Hu, Weicheng
    • Journal of Ginseng Research
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    • v.43 no.4
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    • pp.600-605
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    • 2019
  • Background: The leaves and roots of Panax ginseng are rich in ginsenosides. However, the chemical compositions of the leaves and roots of P. ginseng differ, resulting in different medicinal functions. In recent years, the aerial parts of members of the Panax genus have received great attention from natural product chemists as producers of bioactive ginsenosides. The aim of this study was the isolation and structural elucidation of novel, minor ginsenosides in the leaves of P. ginseng and evaluation of their antiinflammatory activity in vitro. Methods: Various chromatographic techniques were applied to obtain pure individual compounds, and their structures were determined by nuclear magnetic resonance and high-resolution mass spectrometry, as well as chemical methods. The antiinflammatory effect of the new compounds was evaluated on lipopolysaccharide-stimulated RAW 264.7 cells. Results and conclusions: Two novel, minor triterpenoid saponins, ginsenoside $LS_1$ (1) and 5,6-didehydroginsenoside $Rg_3$ (2), were isolated from the leaves of P. ginseng. The isolated compounds 1 and 2 were assayed for their inhibitory effect on nitric oxide production in LPS-stimulated RAW 264.7 cells, and Compound 2 showed a significant inhibitory effect with $IC_{50}$ of $37.38{\mu}M$ compared with that of NG-monomethyl-L-arginine ($IC_{50}=90.76{\mu}M$). Moreover, Compound 2 significantly decreased secretion of cytokines such as prostaglandin $E_2$ and tumor necrosis factor-${\alpha}$. In addition, Compound 2 significantly suppressed protein expression of inducible nitric oxide synthase and cyclooxygenase-2. These results suggested that Compound 2 could be used as a valuable candidate for medicinal use or functional food, and the mechanism is warranted for further exploration.