• Title/Summary/Keyword: Microbial Synthesis

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Microbial production of coenzyme Q10

  • Suh, Jung-Woo
    • 한국약용작물학회:학술대회논문집
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    • 한국약용작물학회 2006년도 Proceedings of The Convention of The Korean Society of Applied Pharmacology
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    • pp.127-130
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    • 2006
  • Coenzyme Q10(CoQ10) is a biological quinine compound that is widely found in living organisms including yeast, plants, and animals. CoQ10 has two major physiological activities:(a)mitochondrial electron-transport activity and (b)antioxidant activity. Various clinical applications are also available : Parkinson's disease, Heart disease, diabetes. Because of its various application filed, the market size of CoQ 10 is continuously expanding all over the world. A Japanese company, Nisshin Pharma Inc. is the first industrial producer of CoQ10(1974). CoQ10 can be produced by fermentation and chemical synthesis. In several companies, these two methods are used for the production of CoQ10:chemical synthesis - Yungjin, Daewoong, Nishin Parma; fermentation - Kaneka, Kyowa, Yungjin, etc. Researchs in microbial production of CoQ10 have several steps: screening of producing microorganisms, strain development, fermentation process, purification process, scale-up process, plant production. Several strategies are available for the strain development : Random mutation and screening, directed metabolic engineering. For the optimization of fermentation process, various conditions (nutrient, aeration, temperature, culture type, etc.) are considered. Purification is one of the most important step because the quality of final products entirely depends on its purity. The production cost will be reduced and the quality of the CoQ10 will be impoved by continuous researches in strain development, fermentation process, purification process.

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Synthesis and Characterization of Fructooligosaccharides Using Levansucrase with a High Concentration of Sucrose

  • Seo Eun-Seong;Lee Jin-Ha;Choi Jae-Young;Seo Mi-Young;Lee Hee-Sun;Chang Seuk-Sang;Lee Hyung-Jong;Choi Jeong-Sik;Kim Doman
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제9권5호
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    • pp.339-344
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    • 2004
  • A method for synthesizing branched fructo-oligosaccharides (BFOS) with a high concentration of sucrose ($1{\~}3$ M) was developed using levansucrase prepared from Leuconortoc mesenteroides B-1355C. The degree of polymerization of oligosaccharides synthesized according to the present method ranged from 2 to over 15. The synthesized BFOS were stable at a pH ranges of 2 to 4 under $120^{\circ}C$. The percentage of BFOS in the reaction digest was $95.7\%$ (excluding monosaccharides; $4.3\%$ was levan). BFOS reduced the insoluble glucan formation by Streptococcus sobrinus on the surfaces of glass vials or stainless steel wires in the presence of sucrose. They also reduced the growth and acid productions of S, sobrinus. Oligosaccharides can be used as sweeteners for foods such as beverages requiring thermo- and acid-stable properties and 3s potential inhibitors of dental caries.

The Effects of Dietary Urea on Microbial Populations in the Rumen of Sheep

  • Kanjanapruthipong, J.;Leng, R.A.
    • Asian-Australasian Journal of Animal Sciences
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    • 제11권6호
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    • pp.661-672
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    • 1998
  • Two experiments were conducted to examine the effects of a range of concentrations of ruminal fluid ammonia ($NH_3$-N) on forage digestibility, microbial growth efficiency and the mix of microbial species. Urea was either continuously infused directly into the rumen of sheep fed 33.3 glh of oaten chaff (Exp. I) or sprayed onto the oaten chaff (750 g/d) given once daily (Exp. 2). Concentrations of $NH_3$-N increased with incremental addition of urea (p < 0.01). Volatile fatty acids (VFA) concentrations and 24 h in sacco organic matter digestibility in the rumen were higher when supplemental urea was given (p < 0.01). The (C2 + C4) : C3 VFA ratio was lower (p < 0.05) when $NH_3$-N was above 200 mgN/I. The fungal sporangia appearing on oat leaf blades were significantly higher when urea was supplemented, indicating that $NH_3$-N was a growthlimiting nutrient for fungi at levels of $NH_3$-N below 30 mgN/l. The density of protozoa was highest when $NH_3$-N concentrations were adjusted to 30 mgN/I for continuously fed ($4.4{\times}10^5/ml$) and to 168 mgN/1 for once daily feeding ($2.9{\times}10^5/ml$). Thereafter increasing concentrations of $NH_3$-N, were associated with a concomitant decline in protozoal densities. At the concentration of $NH_3$-N above 200 mgN/l, the density of protozoa was similar to the density of protozoa in ruminal fluid of the control sheep ($1.8{\times}10^5/ml$). The efficiency of net microbial protein synthesis in the rumen calculated from purine excretion was 17-47% higher when the level of $NH_3$-N was above 200 mgN/1. The possibilities are that 1) there is less bacterial cell lysis in the rumen because of the concomitant decrease in the protozoal pool and/or 2) microbial growth per se in the rumen is more efficient with increasing $NH_3$-N concentrations.

Synthesis of Japanese Encephalitis Virus in Porcine Kidney Stable Cells Observed by Fluorescent Antibody Technique and Autoradiography

  • Lee, Chong-Hoon;Fukai, Konosuke
    • 대한미생물학회지
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    • 제3권1호
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    • pp.51-65
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    • 1968
  • The site of the synthesis of Japanese encephalitis virus(JEV) in the actinomycin-treated and infecter PS Y15 cells(a porcine kidney stable cell line) was observed by the immunofluorescent antibody technique, acridine orange staining, and the autoradiographic analysis. In the parallel studies by immunofluorescent technique and acridine orange staining it the infected cells, Viral protein(as an antigen) and viral RNA were detected at the same site of cytoplasm. In the autoradiographic analysis, the cytoplasmic labeling of $^3H$-uridine was due to the synthesis of JEV-RNA, while the nucleolus and nucleus were not involved. In the autoradiographic studies on the secton of infected cells, the $^3H$-uridine was frequently incorporated around the cytoplasmic vacuoles. This localization of labeling agreed with the site of acridine orange positive granules. The results suggest that the syntheses of the viral RNA and viral protein occurred in the similar site of cytoplasm of the infected cells, and also the virus particles seem to be assembled in the sites of the viral RNA and protein syntheses.

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쑥 Pellet이 면양의 영양소 이용율과 반추위내 발효특성에 미치는 영향 (Effect of Replacing Concentrate with Mugwort Pellet on Nutrient Utilization and Ruminal Fermentation Characteristics in Sheep)

  • 김재황
    • Journal of Animal Science and Technology
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    • 제47권3호
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    • pp.419-428
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    • 2005
  • This study was carried out to investigate the effect of replacing dried mugwort (Artemisia montana Pampan) in concentrate a time of preparation pellet. The treatments, arranged in a 4${\times}$4 Latin square design, were four additional levels of mugwort at 0, 3, 5 and 10% in concentrate. Four crossbred (CorriedalexPolwarth, t) sheep with a mean body weight of 41.3 kg were used to evaluate nutrients digestibility, palatability, fermentation characteristics and microbial protein synthesis in the rumen. The digestibility of crude protein was improved (p < 0.05) to 6.1 % - 8.6 % in sheep fed 3, 5 and 10% mugwort pellet treatments compared with that of control. That of crude fat and NDF was improved (p < 0.05) to 5.8 % - 7.3 % in sheep fed 3 % compared to other treatments. The ruminal pH was significantly (p < 0.05) decreased in sheep fed 3 % mugwort pellet compared to other treatments when observed at 0.5 hour after feeding. The ammonia nitrogen concentrations were the highest in sheep fed all treatments at 1 hour after feeding. The ruminal concentrations of acetic acid and propionic acids were an improvement (p < 0.05) at the 3% and 5% treatments. Retained nitrogen of 3, 5 and 10% treatment with the value of 2.24 - 2.82 g was higher (p < 0.05) than that of the control with 0.78 g and microbial protein production of 10% treatment was higher (p < 0.05) than that of control. This study suggested that the replacing with 3% dry mugwort (Artemisia montana Pampan) in concentrate a time of preparation pellet will improve nutrient digestibility, palatability, ruminal fermentation characteristics and feed value.

Formation of Succinic Acid by Klebsiella pneumoniae MCM B-325 Under Aerobic and Anaerobic Conditions

  • Thakker Chandresh;Bhosale Suresh;Ranade Dilip
    • Journal of Microbiology and Biotechnology
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    • 제16권6호
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    • pp.870-879
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    • 2006
  • The present study describes the formation of succinic acid by a nonvirulent, highly osmotolerant Klebsiella pneumoniae strain SAP (succinic acid producer), its profile of metabolites, and enzymes of the succinate production pathway. The strain produced succinate along with other metabolites such as lactate, acetate, and ethanol under aerobic as well as anaerobic growth conditions. The yield of succinate was higher in the presence of $MgCO_3$ under $N_2$ atmosphere as compared with that under $CO_2$ atmosphere. Analysis of intracellular metabolites showed the presence of a smaller PEP pool than that of pyruvate. Oxaloacetate, citrate, and $\alpha$-ketoglutarate pools were considerably larger than those of isocitrate and fumarate. In order to understand the synthesis of succinate, the enzymes involved in end-product formation were studied. Levels of phosphoenolpyruvate carboxykinase, fumarate reductase, pyruvate kinase, and acetate kinase were higher under anaerobic growth conditions. Based on the profiles of the metabolites and enzymes, it was concluded that the synthesis of succinate took place via oxaloacetate, malate, and fumarate in the strain under anaerobic growth conditions. The strain SAP showed potential for the bioconversion of fumarate to succinate under $N_2$ atmosphere in the presence of $MgCO_3$. At an initial fumarate concentration of 10 g/l, 7.1 g/l fumarate was converted to 7 g/l succinate with a molar conversion efficiency of 97.3%. The conversion efficiency and succinate yield were increased in the presence of glucose. Cells grown on fumarate contained an 18-fold higher fumarate reductase activity as compared with the activity obtained when grown on glucose.

Potential use of Flemingia (Flemingia macrophylla) as a protein source fodder to improve nutrients digestibility, ruminal fermentation efficiency in beef cattle

  • Phesatcha, Burarat;Viennasay, Bounnaxay;Wanapat, Metha
    • Animal Bioscience
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    • 제34권4호
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    • pp.613-620
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    • 2021
  • Objective: This study aimed at studying the potential use of Flemingia (Flemingia macrophylla) as a protein source fodder to improve nutrients digestibility and ruminal fermentation efficiency in beef cattle. Methods: Four, Thai native beef cattle were randomly assigned in a 4×4 Latin square design. Four levels of Flemingia hay meal (FHM) were used to replace soybean meal (SBM) in the concentrate mixtures in four dietary treatments replacing levels at 0%, 30%, 60%, and 100% of SBM. Results: The experimental findings revealed that replacements did not effect on intake of rice straw, concentrate and total dry matter (DM) intake (p>0.05). However, the apparent digestibilities of DM, organic matter, crude protein, acid detergent fiber, and neutral detergent fiber were linearly increased up to 100% replacement levels. Moreover, the production of total volatile fatty acids, and propionate concentration were enhanced (p<0.05) whereas the concentration of acetate was reduced in all replacement groups. Consequently, the CH4 production was significantly lower when increasing levels of FHM for SBM (p<0.05). Furthermore, rumen bacterial population was additionally increased (p<0.05) while protozoal population was clearly decreased (p<0.05) in all replacement groups up to 100%. In addition, microbial nitrogen supply and efficiency of microbial nitrogen synthesis were enhanced (p<0.05), as affected by FHM replacements. Conclusion: The findings under this experiment suggest that 100% FHM replacement in concentrate mixture enhanced rumen fermentation efficiency, nutrients digestibilities, bacterial population, microbial protein synthesis, and subsequently reduced CH4 production in beef cattle fed on rice straw.

Purification, Characterization and Immobilization of Lipase from Proteus vulgaris OR34 for Synthesis of Methyl Oleate

  • Misbah, Asmae;Koraichi, Saad Ibnsouda;Jouti, Mohamed Ali Tahri
    • 한국미생물·생명공학회지
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    • 제48권4호
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    • pp.491-505
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    • 2020
  • A newly isolated strain, Proteus vulgaris OR34, from olive mill waste was found to secrete an alkaline extracellular lipase at 11 U·ml-1 when cultivated on an optimized liquid medium. This lipase was purified 94.64-fold with a total yield of 9.11% and its maximal specific activity was shown to be 3232.58 and 1777.92 U·mg-1 when evaluated using the pH-stat technique at 55℃ and pH 9 and Tributyrin TC4 or olive oil as the substrate. The molecular mass of the pure OR34 lipase was estimated to be around 31 kDa, as revealed by SDS-PAGE and its substrate specificity was investigated using a variety of triglycerides. This assay revealed that OR34 lipase preferred short and medium chain fatty acids. In addition, this lipase was stable in the presence of high concentrations of bile salt (NaDC) and calcium ions appear not to be necessary for its activity. This lipase was inhibited by THL (Orlistat) which confirmed its identity as a serine enzyme. In addition, the immobilization of OR34 lipase by adsorption onto calcium carbonate increased its stability at higher temperatures and within a larger pH range. The immobilized lipase exhibited a high tolerance to organic solvents and retained 60% of its activity after 10 months of storage at 4℃. Finally, the OR34 lipase was applied in biodiesel synthesis via oleic acid mediated esterification of methanol when using hexane as solvent. The best conversion yield (67%) was obtained at 12 h and 40℃ using the immobilized enzyme and this enzyme could be reused for six cycles with the same efficiency.

Synthesis of Chlorogenic Acid and p-Coumaroyl Shikimates from Glucose Using Engineered Escherichia coli

  • Cha, Mi Na;Kim, Hyeon Jeong;Kim, Bong Gyu;Ahn, Joong-Hoon
    • Journal of Microbiology and Biotechnology
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    • 제24권8호
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    • pp.1109-1117
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    • 2014
  • Chlorogenic acid and hydroxylcinnamoyl shikimates are major dietary phenolics as well as antioxidants, with recently discovered biological, activities including protection against chemotheraphy side effects and prevention of cardiovascular disease and cancer. Certain fruits and vegetables produce these compounds, although a microbial system can also be utilized for synthesis of chlorogenic acid and hydroxylcinnamoyl shikimates. In this study, we engineered Escherichia coli to produce chlorogenic acid and p-coumaroyl shikimates from glucose. For the synthesis of chlorogenic acid, two E. coli strains were used; one strain for the synthesis of caffeic acid from glucose and the other strain for the synthesis of chlorogenic acid from caffeic acid and quinic acid. The final yield of chlorogenic acid using this approach was approximately 78 mg/l. To synthesize p-coumaroyl shikimates, wild-type E. coli as well as several mutants were tested. Mutant E. coli carrying deletions in three genes (tyrR, pheA, and aroL) produced 236 mg/l of p-coumaroyl shikimates.

산삼배양액 이용에 관한 반추위 미생물 대사 연구 (Effects of the Artificial Culture Medium of Wild Ginsengs on Rumen Fermentation Characteristics In Vitro)

  • 배귀석;남경표;김혜숙;이상구;최행석;민우기;주종원;맹원재;장문백
    • Journal of Animal Science and Technology
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    • 제45권6호
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    • pp.987-996
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    • 2003
  • 본 실험은 수준별 산삼 배양액에 의한 반추위 내 미생물 발효성상에 미치는 영향에 대하여 조사하기 위해 실시되었다. 산삼 배양액의 수준에 따른 반추위 내 발효성상에 미치는 영향은 WGM을 3% 첨가한 처리구가 대조구 및 다른 WGM 수준 첨가 처리구에 비하여 미생물 단백질 합성량이 가장 높게 나타났다. 산삼 배양액 내에 존재하는 saponin의 영향으로 배양 초기 NH3-N 농도의 수준이 WGM를 첨가한 처리구가 대조구에 비해 낮은 경향(P〈0.05)을 보였으나, 미생물단백질 합성량은 WGM 처리구에서 6시간 이후 급격히 증가(P〈0.05)하는 경향을 나타내었다. 따라서 용해도가 높거나 급여 초기 사료의 이용율이 저하되는 급여 체계에서 산삼 배양액을 첨가는 초기 반추위 미생물의 이용율을 조절할 수 있을 것으로 사료된다. 반추위 내 프로토조아의 수는 WGM 처리구에서 전반적으로 배양 9시간까지 낮아졌고, 3% 처리구에서 가장 낮았는데(P〈0.05), 프로토조아 제거효과에 의해 반추위 미생물합성량 증진시키는 효과를 나타내었다. NDF와 ADF 소화율은 대조구와 모든 처리구에서 배양시간 경과에 따라 높아졌고, NDF 소화율은 대조구와 WGM 3% 처리구에서 그리고 ADF 소화율은 처리구별 차이가 없었다. Total VFA 생성량은 처리구가 대조구에 비해 낮았고, 배양 12시간부터는 5% 처리구에서 차이가 없게 나타났다. 따라서 용해도가 높거나 급여 초기 사료의 이용률이 저하되는 급여 체계에서 산삼 배양액을 첨가는 초기 반추위 미생물의 사료 이용률 증진과 프로토조아 제거효과에 의해 반추위 미생물합성량 증진에 도움이 될 수 있을 것이다.