• Title/Summary/Keyword: Microbial Synthesis

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Effects of Synchronization of Carbohydrate and Protein Supply on Ruminal Fermentation, Nitrogen Metabolism and Microbial Protein Synthesis in Holstein Steers

  • Seo, Ja-Kyeom;Yang, Ji-Young;Kim, Hyun-J.;Upadhaya, Santi Devi;Cho, W.M.;Ha, Jong-K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.11
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    • pp.1455-1461
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    • 2010
  • Three rumen-cannulated Holstein steers were fed three diets, each with a different synchrony index (SI) (LS: 0.77, MS: 0.81, and HS: 0.83), in order to examine the effect of diet on rumen fermentation, nitrogen balance, and microbial protein synthesis. Synchrony index was calculated based on the carbohydrate and crude protein fractions of each ingredient and their degradation rates. Feeding the steers diets with different SIs did not influence dry matter, crude protein, NDF, or ADF digestibility. The concentrations of total and individual VFA in the rumens of steers that were fed the two higher-SI diets were higher than in those fed the low-SI diet (p<0.05), but there was no significant difference between the two higher-SI diets. One hour after feeding, steers on the LS diet had lower ruminal pHs than did those fed the MS or HS diets (p<0.05), and animals on the LS diet generally showed higher ruminal $NH_3$-N levels than did animals on the other diets, with the 4-h post-feeding difference being significant (p<0.05). Steers receiving the LS diet excreted more nitrogen (N) in their urine than did those on the two higher-SI diets (p<0.05), and the total N excretion of those on the LS diet was also higher (p<0.05). Microbial N levels calculated from the concentration of urinary purine derivatives were generally higher when the SI was higher, with the highest microbial protein synthesis being produced by steers on the HS diet (p<0.05). In conclusion, in the current study, ingestion of a synchronous diet by Holstein steers improved microbial protein synthesis and VFA production and decreased total N output.

Effects of Synchronization of Carbohydrate and Protein Supply in Total Mixed Ration with Korean Rice Wine Residue on Ruminal Fermentation, Nitrogen Metabolism and Microbial Protein Synthesis in Holstein Steers

  • Piao, Min Yu;Kim, Hyun-J.;Seo, J.K.;Park, T.S.;Yoon, J.S.;Kim, K.H.;Ha, Jong-K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.11
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    • pp.1568-1574
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    • 2012
  • Three Holstein steers in the growing phase, each with a ruminal cannula, were used to test the hypothesis that the synchronization of the hourly rate of carbohydrate and nitrogen (N) released in the rumen would increase the amount of retained nitrogen for growth and thus improve the efficiency of microbial protein synthesis (EMPS). In Experiment 1, in situ degradability coefficients of carbohydrate and N in feeds including Korean rice wine residue (RWR) were determined. In Experiment 2, three total mixed ration (TMR) diets having different rates of carbohydrate and N release in the rumen were formulated using the in situ degradability of the feeds. All diets were made to contain similar contents of crude protein (CP) and neutral detergent fiber (NDF) but varied in their hourly pattern of nutrient release. The synchrony index of the three TMRs was 0.51 (LS), 0.77 (MS) and 0.95 (HS), respectively. The diets were fed at a restricted level (2% of the animal's body weight) in a $3{\times}3$ Latin-square design. Synchronizing the hourly supply of energy and N in the rumen did not significantly alter the digestibility of dry matter, organic matter, crude protein, NDF or acid detergent fiber (ADF) (p>0.05). The ruminal $NH_3$-N content of the LS group at three hours after feeding was significantly higher (p<0.05) than that of the other groups; however, the mean values of ruminal $NH_3$-N, pH and VFA concentration among the three groups were not significantly different (p>0.05). In addition, the purine derivative (PD) excretion in urine and microbial-N production (MN) among the three groups were not significantly different (p>0.05). In conclusion, synchronizing dietary energy and N supply to the rumen did not have a major effect on nutrient digestion or microbial protein synthesis (MPS) in Holstein steers.

Production of L-DOPA by Thermostable Tyrosine Phenol-lyase of a Thermophilic Symbiobacterium Species Overexpressed in Recombinant Escherichia coli

  • Lee, Seung-Goo;Ro, Hyeon-Su;Hong, Seung-Pyo;Kim, Eun-Hwa;Sung, Moon-Hee
    • Journal of Microbiology and Biotechnology
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    • v.6 no.2
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    • pp.98-102
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    • 1996
  • A thermostable tyrosine phenol-lyase gene of a thermophilic Symbiobacterium species was cloned and overexpressed in Escherichia coli in order to produce the biocatalyst for the synthesis of 3, 4-dihy-droxyphenyl-L-alanine (L-DOPA). The substrates used for the synthetic reaction were pyrocatechol, so-dium pyruvate, and ammonium chloride. The enzyme was stable up to $60^{\circ}C$, and the optimal temperature for the synthesis of L-DOPA was $37^{\circ}C$ . The optimal pH of the reaction was about 8.3. Enzyme activity was highly dependent on the amount of ammonium chloride and the optimal concentration was estimated to be 0.6 M. In the case of pyrocatechol, an inactivation of enzyme activity was observed at con-centrations higher than 0.1 M. Enzyme activity was increased by the presence of ethanol. Under op-timized conditions, L-DOPA production was carried out adding pyrocatechol and sodium pyruvate to the reaction solution intermittently to avoid substrate depletion during the reaction. The concentration of L-DOPA reached 29.8 g/l after 6 h, but the concentration didn t increase further because of the formation of byproducts by a non-enzymatic reaction between L-DOPA and pyruvate.

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Effect of Levels of Supplementation of Concentrate Containing High Levels of Cassava Chip on Rumen Ecology, Microbial N Supply and Digestibility of Nutrients in Beef Cattle

  • Wanapat, M.;Khampa, S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.1
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    • pp.75-81
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    • 2007
  • The object of this study was to determine the influence of supplementation of concentrate containing high levels of cassava chip on rumen ecology, microbial protein and digestibility of nutrients. Four, rumen fistulated crossbred beef steers with initial body weight of 400${\pm}$10 kg were randomly assigned according to a 4${\times}$4 Latin square design. The dietary treatments were concentrate cassava chip based offering at 0, 1, 2 and 3% BW with urea-treated rice straw fed ad libitum. It was found that ruminal pH was significantly decreased with increase of concentrate. Volatile fatty acids (VFA) concentration in the rumen was significantly different among treatments. In addition, a molar proportion of propionate was higher in supplemented groups at 2 and 3% BW (p<0.05), leading to significantly decreased acetate:propionate ratio. Furthermore, microbial N supply was significantly improved and was highest at 2% BW supplementation. The efficiency of rumen microbial-N synthesis based on organic matter (OM) truly digested in the rumen was highest in level of concentrate supplementation at 2% BW (80% of cassava chip in diets). Moreover, bacterial populations such as amylolytic bacteria was linearly increased, while cellulolytic bacteria was linearly decreased (p<0.01) when cattle received concentrate supplementation in all levels. The total protozoal counts were significantly increased, while fungal zoospores were dramatically decreased in cattle receiving increased levels of concentrate. In conclusion, cassava chip can be use as energy source at 80% in concentrate and supplementation of concentrate at 2% BW with urea-treated rice straw as roughage could improve rumen fermentation efficiency in beef cattle.

The Requirement of Ruminal Degradable Protein for Non-Structural Carbohydrate-Fermenting Microbes and Its Reaction with Dilution Rate in Continuous Culture

  • Meng, Q.X.;Xia, Z.G.;Kerley, M.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.10
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    • pp.1399-1406
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    • 2000
  • A continuous culture study was conducted to determine the impact of ruminal degradable soy protein (S-RDP) level and dilution rate (D) on growth of ruminal non-structural carbohydrate-fermenting microbes. Corn starch, urea and isolated soy protein (ISP) were used to formulate three diets with S-RDP levels of 0, 35 and 70% of total dietary CP. Two Ds were 0.03 and $0.06h^{-1}$ of the fermenter volume in a single-effluent continuous culture system. As S-RDP levels increased, digestibilities of dietary dry matter (DM), organic matter (OM) and crude protein (CP) linearly (p=0.001) decreased, whereas digestion of dietary starch linearly (p=0.001) increased. Increasing D from 0.03 to $0.06h^{-1}$ resulted in decreased digestibilities of dietary DM and OM, but had no effect on digestibilities of dietary starch (p=0.77) and CP (p=0.103). Fermenter pH, the concentration of volatile fatty acids (VFA) and daily VFA production were unaffected (p=0.159-0.517) by S-RDP levels. Molar percentages of acetate, propionate and butyrate were greatly affected by S-RDP levels (p=0.016-0.091), but unaffected by D (p=0.331-0.442). With increasing S-RDP levels and D, daily bacterial counts, daily microbial N production (DMNP) and microbial efficiency (MOEFF; grams of microbial N produced per kilogram of OM truly digested) were enhanced (p=0.001). The increased microbial efficiency with increasing S-RDP levels is probably the result of peptides or amino acids that served as a stimulus for optimal protein synthesis. The quantity of ruminal degradable protein from soy proteins required for optimum protein synthesis of non-structural carbohydrate-fermenting microbes appears to be equivalent to 9.5% of dietary fermented OM.

Potential Industrial Applications and Evolution of Carbohydrolases and Glycansucrases

  • Kim, Do-Man;Seo, Eun-Seong;Lee, Jin-Ha;Kang, Hee-Kyoung;Cho, Jae-Young
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2004.06a
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    • pp.215-218
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    • 2004
  • Dextrans make up a class of polysaccharides that are D-glucans of various structures with contiguous $\alpha$-1longrightarrow6 ~6 glycosidic linkages in the main chains and $\alpha$-1longrightarrow2, $\alpha$-1longrightarrow3, or $\alpha$-1longrightarrow4 branch glycosidic linkages, depending on the specificity of the particular dextransucrase. Glucansucrases that catalyze glucans synthesis from sucrose. When other carbohydrates, in addition to sucrose, are present in the enzyme digest, the enzyme transfers glucose to the carbohydrate acceptors in the secondary reaction that diverts some of the glucose from incorporation into glucan. Many carbohydrate acceptors have been recognized and the products that result are dependent on the particular enzyme and the structure of the particular acceptor. Because of these unique catalytic characteristics, various dextransucrases have many important industrial and medical uses. To improve the understanding of their action mode and extend their applications, this study describes mechanism of glucan synthesis and potential industrial uses of dextransucrases, and our recent findings on the structural, functional organization and directed evolution of the glucansucrases to offer for designing glucansucrases with improved properties.

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Anti-Microbial and Anti-Wrinkle Effect of Kaempferol and Kaempferol Rhamnosides isolated from Hibiscus cannabinus L. (양마에서 분리한 Kaempferol과 그 배당체의 항균 및 주름억제 효과)

  • Lee, Keun Ha;Kong, Hae Jin;Cho, Young Long;Joo, Chul Gue;Kwon, Sun Sang;Hwang, Jae Sung;Park, Chung
    • Korean Journal of Medicinal Crop Science
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    • v.20 no.6
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    • pp.454-460
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    • 2012
  • In this study, kaempferol and its rhamnosides (${\alpha}$-rhamnoisorobin, afzelin, kaempferitrin) were isolated electively by bioconversion technology from Hibiscus cannabinus L. leaves to evaluate the anti-wrinkles effects and anti-microbial effects. In order to evaluate anti-wrinkles activity, reduction of expression matrix metalloproteinase-1 (MMP-1) protein and proliferation/ pro-collagen production were investigated. Kaempferol and ${\alpha}$-rhamnoisorobin showed inhibition activity of MMP-1 generated to compared to positive control. In HaCaT cell proliferation assay, kaempferol and ${\alpha}$-rhamnoisorobin significantly promoted cell proliferation in a concentration-dependent manner. In addition, procollagen synthesis assays (by HDF-N cell) showd that TGF-${\beta}$ induced procollagen production and also, all four kinds of experimental significantly promoted procollagen synthesis in a concentration-dependent manner. Kaempferol and ${\alpha}$-rhamnoisorobin exhibited strong antimicrobial activities on five of microbes, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Candida albicans and Aspergillus niger.

Downregulation of EHT1 and EEB1 in Saccharomyces cerevisiae Alters the Ester Profile of Wine during Fermentation

  • Yang, Xue;Zhang, Xuenan;He, Xi;Liu, Canzhen;Zhao, Xinjie;Han, Ning
    • Journal of Microbiology and Biotechnology
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    • v.32 no.6
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    • pp.761-767
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    • 2022
  • EHT1 and EEB1 are the key Saccharomyces cerevisiae genes involved in the synthesis of ethyl esters during wine fermentation. We constructed single (Δeht1, Δeeb1) and double (Δeht1Δeeb1) heterogenous mutant strains of the industrial diploid wine yeast EC1118 by disrupting one allele of EHT1 and/or EEB1. In addition, the aromatic profile of wine produced during fermentation of simulated grape juice by these mutant strains was also analyzed. The expression levels of EHT1 and/or EEB1 in the relevant mutants were less than 50% of the wild-type strain when grown in YPD medium and simulated grape juice medium. Compared to the wild-type strain, all mutants produced lower amounts of ethyl esters in the fermented grape juice and also resulted in distinct ethyl ester profiles. ATF2, a gene involved in acetate ester synthesis, was expressed at higher levels in the EEB1 downregulation mutants compared to the wild-type and Δeht1 strains during fermentation, which was consistent with the content of acetate esters. In addition, the production of higher alcohols was also markedly affected by the decrease in EEB1 levels. Compared to EHT1, EEB1 downregulation had a greater impact on the production of acetate esters and higher alcohols, suggesting that controlling EEB1 expression could be an effective means to regulate the content of these aromatic metabolites in wine. Taken together, the synthesis of ethyl esters can be decreased by deleting one allele of EHT1 and EEB1 in the diploid EC1118 strain, which may modify the ester profile of wine more subtly compared to the complete deletion of target genes.

Synergistic Effect of Reductase and Keratinase for Facile Synthesis of Protein-Coated Gold Nanoparticles

  • Gupta, Sonali;Singh, Surinder P.;Singh, Rajni
    • Journal of Microbiology and Biotechnology
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    • v.25 no.5
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    • pp.612-619
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    • 2015
  • We have synthesized gold nanoparticles (GNPs) using chicken feathers (poultry waste) and Bacillus subtilis RSE163. Disulfide reductase and keratinase produced by Bacillus subtilis during the degradation of chicken feather has been used to reduce Au3+ from HAuCl4 precursor to produce gold nanoparticles. The synthesized biogenic GNPs were characterized by UV-visible spectroscopy, transmission electron microscopy (TEM), and zeta potential measurements. Fourier transform infrared (FTIR) spectroscopy indicated the presence of protein capping on synthesized GNPs, imparting multifunctionality to the GNP surface. Furthermore, the nontoxic nature of biogenic GNPs was insured by interaction with Escherichia coli (ATCC11103), where TEM images and enhancement of growth rate of E. coli in log phase signified their nontoxic nature. The results indicate that the synthesis of biocompatible GNPs using poultry waste may find potential applications in drug delivery and sensing.

Feeding Value of Ammoniated Rice Straw Supplemented with Rice Bran in Sheep : I. Effects on Digestibility, Nitrogen Retention and Microbial Protein Yield

  • Orden, E.A.;Yamaki, K.;Ichinohe, T.;Fujihara, T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.4
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    • pp.490-496
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    • 2000
  • In vivo digestibility, nitrogen retention and microbial protein yield from diets of 100% ammonia treated rice straw (ARS) ($D_1$); 65% untreated rice straw (URS)+30% rice bran (RB)+5% SBM ($D_2$) and 85% ARS+15% RB ($D_3$) were determined using three Japanese Corriedale wethers in a $3{\times}3$ Latin Square Design. Results showed that DM consumption and organic matter digestibility were highest in $D_3$; but did not promote high protein digestibility, which RB+SBM had effected in URS based-diet. Dry matter intake and OM digestibility were the same for $D_1$ and $D_3$. Solubility of fiber bonds was increased by ammoniation, resulting in higher NDF digestibility. Nitrogen retention and microbial protein yield of rice bran supplemented groups was higher than ARS, but supplementation did not significantly increase efficiency of microbial protein synthesis from ARS which did occur when RB+SBM was added to untreated straw. The quality of ammoniated rice straw could be improved through RB supplementation because of its positive effects on DM digestibility, nitrogen retention and microbial protein yield. However, the addition of RB+SBM to URS resulted to more efficient N utilization.