• Title/Summary/Keyword: TRYPTONE

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Liquid culture condition of Tremella fuciformis mycelia (흰목이 균사 액체배양 조건)

  • Chang, Hyun-You;Lee, Chan;Choi, Sung-Woo;Yun, Jong Won
    • Journal of Mushroom
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    • v.6 no.1
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    • pp.27-31
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    • 2008
  • The optimization of submerged culture conditions for mycelial growth and exopolysaccharide (EPS) production in an edible mushroom Tremella fuciformis were studied in shake flasks and bioreactors. The temperature of $28^{\circ}C$ and pH 8 in the beginning of fermentation in agitated flasks was the most efficient condition to obtain maximum mycelial biomass and EPS. The optimal medium constituents were as follows (g l-1): glucose 20, tryptone 2, $KH_2PO_4$ 0.46, $K_2HPO_4$ 1 and $MgSO_4H_2O$ 0.5. The fungus was cultivated under various agitation and aeration conditions in a 5L stirred-tank bioreactor. The maximum cell mass and EPS production were obtained at a relatively high agitation speed of 200 rpm and at an aeration rate of 2 vvm. The flow behavior of the fermentation broth was Newtonian and the maximum apparent viscosity (35 cP) was observed at a highly aerated condition (2 vvm). The EPS productivity in an airlift reactor was higher than that in the stirred-tank reactor. The EPS was protein-bound polysaccharides consisted of mainly mannose, xylose, and fructose. The molecular weights of EPS were determined to be $1.3{\sim}1.5{\times}10^6$.

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Cultural Conditions for Mycelial Growth and Molecular Phylogenetic Relationship in Different Wild Strains of Schizophyllum commune

  • Alam, Nuhu;Cha, Youn-Jeong;Shim, Mi-Ja;Lee, Tae-Soo;Lee, U-Youn
    • Mycobiology
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    • v.38 no.1
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    • pp.17-25
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    • 2010
  • The common split-gilled mushroom, Schizophyllum commune is found throughout the world on woody plants. This study was initiated to evaluate conditions for favorable vegetative growth and to determine molecular phylogenetic relationship in twelve different strains of S. commune. A suitable temperature for mycelial growth was obtained at $30^{\circ}C$. This mushroom grew well in acidic conditions and pH 5 was the most favorable. Hamada, glucose peptone, Hennerberg, potato dextrose agar and yeast malt extract were favorable media for growing mycelia, while Lilly and glucose tryptone were unfavorable. Dextrin was the best and lactose was the less effective carbon source. The most suitable nitrogen sources were calcium nitrate, glycine, and potassium nitrate, whereas ammonium phosphate and histidine were the least effective for the mycelial growth of S. commune. The genetic diversity of each strain was investigated in order to identify them. The internal transcribed spacer (ITS) regions of rDNA were amplified using PCR. The size of the ITS1 and ITS2 regions of rDNA from the different strains varied from 129 to 143 bp and 241 to 243 bp, respectively. The sequence of ITS1 was more variable than that of ITS2, while the 5.8S sequences were identical. A phylogenetic tree of the ITS region sequences indicated that the selected strains were classified into three clusters. The reciprocal homologies of the ITS region sequences ranged from 99 to 100%. The strains were also analyzed by random amplification of polymorphic DNA (RAPD) with 20 arbitrary primers. Twelve primers efficiently amplified the genomic DNA. The number of amplified bands varied depending on the primers used or the strains tested. The average number of polymorphic bands observed per primer was 4.5. The size of polymorphic fragments was obtained in the range of 0.2 to 2.3 kb. These results indicate that the RAPD technique is well suited for detecting the genetic diversity in the S. commune strains tested.

Medium Optimization for Pediocin SA131 Production by Pediococcus pentosaceus SA131 against Bovine Mastitis Using Response Surface Methodology

  • Park, Yeo-Lang;Lee, Na-Kyoung;Park, Keun-Kyu;Park, Yong-Ho;Kim, Jong-Man;Nam, Hyang-Mi;Jung, Suk-Chan;Paik, Hyun-Dong
    • Food Science of Animal Resources
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    • v.30 no.1
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    • pp.66-72
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    • 2010
  • Pediococcus pentosaceus SA131 was isolated from jeotgal, is the bacteriocin producer against bovine mastitis pathogens, Streptococcus uberis E290, Enterococcus gallinarum E362, and Staphylococcus epidermidis ATCC 12228. The medium composition for pediocin SA131 production by P. pentosaceus SA131 was optimized using response surface methodology. Component of medium was studied as carbon source (glucose, fructose, lactose, glycerol, sucrose, maltose, and mannitol), nitrogen source (beef extract, yeast extract, peptone, malt extract, and tryptone), mineral and surfactant ($MgSO_4$, $KH_2PO_4$, $(NH_4)_2SO_4$, $MnSO_4$, NaCl, sodium acetate, and Tween 80). Through one factor-at-a-time experiment, glucose, fructose, yeast extract, malt extract, NaCl, $MgSO_4$, and Tween 80 were determined as the good ingredient. The effects of major factors for pediocin SA131 production were investigated by two-level fractional factorial designs (FFD). By a $2^4$ FFD, fructose, yeast extract, and $MnSO_4$ were found to be the important factors for the bacteriocin production. Subsequently, a $2^3$ central composite design (CCD) was adopted to derive a statistical model for optimizing the composition of the fermentation medium. The estimated optimum composition for the production of pediocin SA131 by P. pentosaceus SA131 was as follows; 0.13% fructose, 1% glucose, 1.8% yeast extract, 2.58% $MnSO_4$, 0.2% NaCl, and 0.2% Tween 80. The pediocin production under optimized medium was increased to 1,000 AU/mL, compared to the 400 AU/mL in MRS medium.

Studies on Microbial Penicillin Amidase ( I ) Optimization of the Enzyme Production from Escherichia coli (미생물 페니실린 아미다제에 관한 연구 (I) E. coli로부터 효소생산 조건의 최적화)

  • Kim, Bong-Hee;Seong, Baik-Lin;Mheen, Tae-Iek;Moon H. Ban
    • Microbiology and Biotechnology Letters
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    • v.9 no.1
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    • pp.29-34
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    • 1981
  • To maximize the production of penicillin amidase from Estherichia coli (ATCC 9637), the media composition and several factors affecting the engyme production during fermentation were studied. The optimal media composition was found to be; 3.5% tryptone, 1.5% monosodium glutamate and 0.5% yeast extract. The addition of 0.15% phenylacetic acid as an enzyme inducer at the initial stage of cultivation increased the engyme productivity about 5 fold. It was found that the engyme activity reached maximum within 16hr of cultivation. The maximum production of the enzyme obtained was about 102.5 units/l broth under the optimized condition. The enzyme production was markedly increased by the optimization as compared with those previously reported.

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Detection of Polyhydroxyalkanoate-Accumulating Bacteria from Domestic Wastewater Treatment Plant Using Highly Sensitive PCR Primers

  • Huang, Yu-Tzu;Chen, Pi-Ling;Semblante, Galilee Uy;You, Sheng-Jie
    • Journal of Microbiology and Biotechnology
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    • v.22 no.8
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    • pp.1141-1147
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    • 2012
  • Polyhydroxyalkanoate (PHA) is a class of biodegradable plastics that have great potential applications in the near future. In this study, the micro-biodiversity and productivity of PHA-accumulating bacteria in activated sludge from a domestic wastewater treatment plant were investigated. A previously reported primer set and a self-designed primer set (phaCF1BO/phaCR2BO) were both used to amplify the PHA synthase (phaC) gene of isolated colonies. The new primers demonstrated higher sensitivity for phaC, and combining the PCR results of the two primer sets was able to widen the range of detected genera and raise the sensitivity to nearly 90%. Results showed that 85.3% of the identified bacteria were Gram-negative, with Ralstonia as the dominant genus, and 14.7% were Gram-positive. In addition, Zoogloea and Rhizobium contained the highest amounts of intracellular PHA. It is apparent that glucose was a better carbon source than pentone or tryptone for promoting PHA production in Micrococcus. Two different classes, class I and class II, of phaC were detected from alphaproteobacteria, betaproteobacteria, and gammaproteobacteria, indicating the wide diversity of PHA-accumulating bacteria in this particular sampling site. Simultaneous wastewater treatment and PHA production is promising by adopting the high PHA-accumulating bacteria isolated from activated sludge.

Medium Optimization and Application of Affinity Column Chromatography for Trypsin Production from Recombinant Streptomyces griseus

  • Chi, Won-Jae;Song, Ju-Hyun;Oh, Eun-A.;Park, Seong-Whan;Chang, Yong-Keun;Kim, Eung-Soo;Hong, Soon-Kwang
    • Journal of Microbiology and Biotechnology
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    • v.19 no.10
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    • pp.1191-1196
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    • 2009
  • The production of Streptomyces griseus trypsin (SGT) by S. griseus IFO13350 transformed with the expression vector pWHM3-TR1R2, containing sprT encoding SGT and the two positive regulatory genes sgtR1 and sgtR2, was investigated in various media. Cultivation in Ferm-0 gave 1.4 times more trypsin activity than in C5/L medium. In addition, replacement of 2% glucose and 1% skim milk in Ferm-0 with 2% dextrin and 1% tryptone (designated Ferm-II) enhanced trypsin activity 4.1-fold. To simplify the purification process, the supernatant from the S. griseus transformant cultured in Ferm-II medium was fractionated with ammonium sulfate (25-55%), then subjected to Hitrap Benzamidine FF affinity column chromatography. The specific activity of SGT purified by one-step chromatography was 69,550 unit/mg protein and the overall purification yield was above 8%, indicating that this method is more effective than those previously reported. Purified SGT was most active at pH 8.0 and $50^{\circ}C$, and it maintained activity between pH 7.0 and 9.0 and at temperatures up to $70^{\circ}C$. These enzymatic properties are very similar to those of authentic eukaryotic trypsin purified from bovine pancreas.

Thermococcus onnurineus sp. nov., a Hyperthermophilic Archaeon Isolated from a Deep-Sea Hydrothermal Vent Area at the PACMANUS Field

  • Bae, Seung-Seob;Kim, Yun-Jae;Yang, Sung-Hyun;Lim, Jae-Kyu;Jeon, Jeong-Ho;Lee, Hyn-Sook;Kang, Sung-Gyun;Kim, Sang-Jin;Lee, Jung-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.16 no.11
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    • pp.1826-1831
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    • 2006
  • A novel hyperthermophilic, anaerobic, heterotrophic archaeon, designated strain $NA1^T$, was isolated from a deep-sea hydrothermal vent area (depth, 1,650 m) within the Papua New Guinea-Australia-Canada-Manus (PACMANUS) field. Cells of this strain were motile by means of polar flagella, coccoid-shaped with a diameter of approximately $0.5-1.0{\mu}m$, and occurred as single cells. Optimal temperature, pH, and NaCl concentration for growth were $80^{\circ}C$, 8.5, and 3.5%, respectively. The new isolate was an obligate heterotroph that utilized yeast extract, beef extract, tryptone, peptone, casein, and starch as carbon and energy sources. Elemental sulfur was required for growth and was reduced to hydrogen sulfide. The G+C content of the genomic DNA was 52.0 mol%. Phylogenetic analysis of the 16S rRNA gene indicated that strain $NA1^T$ belongs to the genus Thermococcus, and the organism is most closely related to T. gorgonarius, T. peptonophilus, and T. celer; however, no significant homology was observed among species by DNA-DNA hybridization. Strain $NA1^T$ therefore represents a novel species for which the name Thermococcus onnurineus sp. novo is proposed. The type strain is $NA1^T$ (=KCTC 10859, =JCM 13517).

Production of HCN, Weed Control Substance, by Pseudomonas koreensis and its Plant Growth-Promoting and Termiticidal Activities (Pseudomonas koreensis에 의한 잡초제어활성물질인 HCN 생성과 이 균주의 식물성장 촉진 및 흰개미 살충 활성)

  • Yoo, Ji-Yeon;Jang, Eun-Jin;Park, Soo-Yeun;Son, Hong-Joo
    • Journal of Environmental Science International
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    • v.27 no.9
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    • pp.771-780
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    • 2018
  • To develope a microbial weed control agent, HCN-producing bacteria were isolated, and their characteristics were investigated. A selected strain of WA15 was identified as Pseudomonas koreensis by morphological, cultural, biochemical and 16S rRNA gene analyses. The conditions for HCN production was investigated by a One-Variable-at-a-Time (OVT) method. The optimal HCN production conditions were tryptone 1%, glycine 0.06%, NaCl 1%, and an initial pH and temperature of 5.0 and $30^{\circ}C$, respectively. The major component for HCN production was glycine. Under optimal conditions, HCN production was about 3 times higher than that of the basal medium. The WA15 strain had physiological activities, such as indoleacetic acid that was associated with the elongation of plant roots and siderophore and ammonification inhibiting fungal growth, and produced hydrolytic enzymes, such as cellulase, pectinase and lipase. The strain was able to inhibit the growth of phytopathogenic fungi, such as Rhizoctonia solani, Botrytis cinerea and Fusarium oxysporum, by the synergistic action of volatile HCN and diffusible antimicrobial compounds. A microscopic observation of R. solani that was teated with the WA15 strain showed morphological abnormalities of fungal mycelia, which could explain the role of the antimicrobial metabolites that were produced by the WA15 strain. The volatile HCN produced by the WA15 strain was also found to have insecticidal activity against termites. Our results indicate that Pseudomonas koreensis WA15 can be applied as a microbial agent for weed control and also as a termite repellent. Furthermore, it could be applied as a microbial termiticidal agent to replace synthetic insecticides.

Production of Carotenoid from Halophilic Erythrobacter sp. and characterization of Physiological Properties (해양미생물 Erythrobacter 속으로부터 Carotenoid의 생산 및 그 생리활성)

  • 김종덕;강동수;김민용;노승배;최명락;송상호;백승한;서효진;김대현
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.1
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    • pp.143-151
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    • 2001
  • A marine bacterium producing carotenoid was isolated from the Yosu coastal area of South Korea, which was recorded as MCK-1. It was identified as Erythrobacter sp. Optimium conditions of marine carotenoid fermentation from Erythrobacter sp. were pH 6.0, a temperature of $25^{\circ}C$, 16 mM mannitol as a carbon source, 0.5% tryptone as a nitrogen source, 0.1 mM $Fe^{+2}$ ion as a mineral source and 1$\mu$M of cyanocobalamine as a growth factor in a jar-fermentor. Erythrobacter sp. was produced 351.27 mg/100mL of the marine carotenoid in these optimum conditions. This marine carotenoid was composed of 4 different conpounds, like as notoxanthin (61.4%), can thaxanthin (24.6%), fucoxanthin (8.2%), and zeaxanthin (5.8%). Physiological properties including antibacterial activity, cytotoxic effect, antioxidative effect and free radical scavenging activity were characterized with crude carotenoid. Carotenoid exhibited no antibacterial activity against E. coli and lactobacillus bulgaricus, but showed cytotoxic effect against cancer cells such as HepG2 (Hepatocellular carcinoma, human, ATCC HB-8065) and HeLa (Cervical carcinoma, human, ATCC CCL-2) cells. The impediment ratios for HepG2 and HeLa cell were 37.14% and 33.78%, respectively. This carotenoid expressed a strong antioxidative effect (77%) against CCL-13 5 $\mu\textrm{g}$/mL and 50 $\mu\textrm{g}$/mL crude carotenoid, respectively.

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Effects of Culture Condition on Solubilization of Coal by Microorganisms (배양 조건의 변화가 미생물에 의한 석탄의 액화에 미치는 영향)

  • 이현호;신현재양지원
    • KSBB Journal
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    • v.11 no.4
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    • pp.462-469
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    • 1996
  • Biosolubilization of an Australian lignite was investigated by using Streptomyces viridosporus and Poria cocos. In order to solubilize coals effectively they were pretreated by nitric acid both in surface and liquid cultures. The optimum growth pH was 7.5 for S. viridosporus and 4.5 for P. cocos. The effects of various carbon, nitrogen and metal sources on overall solubilization were also studied. Solubility increased with the addition of urea for S. viridosporus, and peptone and tryptone for P. cocos. However carbon and metal sources had little or negative effects on solubilization. Maximum amount of coal solubilized was 85%(w/w) in a batch fermentation culture. Extracellular materials produced by micro-organism were found to be responsible for the coal solubilization. Approximately 70 to 80% of coal solubilization was determined to be the result of non-enzymatic reactions, and the rest to be the result of enzymatic reactions. Characteristics of the solubilized coal were compared with those of original coal and pretreated coal by the approximate and ultimate composition analysis, and IR-spectrum analysis. The spectroscopic results showed that the mechanism of coal solubilization was caused by continuous oxidation.

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