• Title/Summary/Keyword: shaking culture

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Characteristics and purification of proteoglycan from Phellinus igniarius (Phellinus igniarius로부터 분리한 단백다당류의 분리 및 특성)

  • Kim, Seon-Hee;Jung, In-Chang;Kwon, Yong-Il;Kim, So-Yeun;Lee, Jong-Suk;Lee, Hang-Woo;Lee, Jae-Sung
    • Applied Biological Chemistry
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    • v.43 no.1
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    • pp.57-62
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    • 2000
  • The proteoglycan, intracellular and extracellular, extracted from the liquid culture of Phellinus igniarius were purified and characterized. The mycelial productivity was proved to be better in shaking culture compared to standing culture. The productivity of intracellular proteoglycan of Phellinus igniarius appeared to be similar in two culturing methods. The standing culture of Phellinus igniarius produced 6 times as much extracellular proteoglycan compared to shaking culture. The proteoglycan were purified to a single peak by ion exchange chromatography(DEAE-cellulose) followed by gel filtration(Sepharose 2B). PIEPDG contained 79.0% total sugar and 7.2% protein. PIEPAG contained 56.7% total sugar and 40.8% protein. PIIPDG contained 64.8% total sugar and 17.4% protein. PIIPAG contained 56.9% total sugar and 41.5%n protein. The molecular weights of all the fractions were estimated to be above 100,000, from 134KDa of PIEPDG to 560 KDa of PIEPAG. The results of sugar analysis by HPLC showed that PIEPDG contains glucose only. The sugar part of PIIPDG and PIIPAG were consisted of glucose and inositol. The PIEPAG contained three kinds of monosaccharides, glucose, fructose and inositol.

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Favorable Condition for Mycelial Growth of Tricholoma matsutake (송이균 배양을 위한 균사생장 조건)

  • Kim, In-Yeup;Jung, Gwang-Reul;Han, Sang-Kuk;Cha, Joo-Young;Sung, Jae-Mo
    • The Korean Journal of Mycology
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    • v.33 no.1
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    • pp.22-29
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    • 2005
  • The main objectives of this research were to study the cultural and nutritional characteristics of Tricholoma matsutake and to establish its liquid culture system. The optimum growth of T. matsutake was observed in HA and TMM agar media. Similarly highest growth was observed in PDB and TMM liquid media. The optimal temperature for the mycelial growth was $25^{\circ}C$. The most suitable carbon source was dextrin among 12 different carbon sources tested. Yeast extract and peptone were best nitrogen sources among 17 different sources tested. The optimum mineral salts were $Fe_{2}(SO_{4})_{3}{\cdot}H_{2}O$ and KCl among 9 different sources tested. Shaking culture gave higher mycelial growth compared to stationary culture. Similarly, optimum medium amount for shaking culture was 100 ml per 250 ml flask. The highest mycelial growth was obtained when $5{\sim}7$ mycelial discs were inoculated in 100 ml of medium and incubated for $8{\sim}9$ weeks, respectively. The highest proportion of mycelial growth was observed at 40 : 1 ratio of medium to inoculum volume in 8 l air-lift fermenter.

Bioluminescent Determination of Lactose Secretion: A Measure of the In Vitro Performance of Mammary Acini from Lactating Rats

  • Choi, B.H.;Stewart, K.W.;Davis, S.R.;Myung, K.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.2
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    • pp.274-278
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    • 2002
  • A culture system for lactating rat mammary acini was evaluated, where the primary indicator of performance was lactose secretion, measured by a sensitive bioluminescence assay. Lactose secretion was reduced by half (p<0.01) over the first 6 h of culture by overnight feed withdrawal (FW) from tissue donors but was sensitive to increased glucose concentration in the culture media (p<0.001) up to 30 mM. Lactose production of cells from fed donors over the first 6 h in culture in 30 mM glucose was 8.9 fmol/cell/h - a rate calculated to be about half that in vivo. No significant difference was shown in lactose secretion by cells from fed or FW rats over 6-24 h. Lactose secretion was 3.6 fmol/cell/h by cells from fed animals in 40 mM glucose concentration media over the 6-24 h culture period. Addition of insulin to the culture media had no effect on rates of lactose secretion while addition of prolactin and hydrocortisone, with or without insulin, significantly (p<0.001) decreased lactose production over both 0-6 h and 6-24 h culture periods. Lactose synthesis in vitro was significantly enhanced by aeration of the media during collagenase digestion of mammary tissue (p<0.05). No improvement in lactose secretion was effected by shaking of cells during culture, Matrigel coating of culture dishes or change in cell density over a range up to 2.5 million cells per ml.

Production of Bacterial Cellulose by Gluconacetobacter hansenii PJK Isolated from Rotten Apple

  • Park, Joong-Kon;Park, Youn-Hee;Jung, Jae-Yong
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.8 no.2
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    • pp.83-88
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    • 2003
  • A cellulose-producing strain isolated from rotten apples was identified as Gluconacetobacter hansenii based on its physiological properties and the 16S rDNA complete sequencing method, and specifically named Gluconacetobacter hansenii PJK. The amount of bacterial cellulose (BC) produced by G. hansenii PJK in a shaking incubator was 1.5 times higher than that produced in a static culture. The addition of ethanol to the medium during cultivation enhanced the productivity of bacterial cellulose, plus the supplementation of 1% ethanol into the culture medium made the produced BC aggregate into a big lump and thus protected the bacterial-cellulose-producing G. hansenii PJK cells in the shear stress field from being converted into non-cellulose-producing (Cel) mutants. Cells subcultured three times in a medium containing ethanol retained their ability to produce BC without any loss in the production yield.

재조합 효모를 이용한 endoinulinase의 생산 특성

  • Han, Ji-Hye;Lee, Eun-Mi;Yun, Yeong-Mi;Lee, Hyeon-Cheol;Jeong, Bong-U;Chae, Geon-Sang
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.478-481
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    • 2000
  • The INU2 gene encoding an endoinulinase of Aspergillus ficuum was expressed by the Kluyveromyces marxianus INU1 promoter in a SUC2-deleted Saccharomyces cerevisiae to produce the endoinulinase free of an exoinulinase and an extracellular invertase in the culture medium. When inulin was included in the medium, a recombinant yeast strain produced the sufficient amount of the enzyme to make a halo around its colony. An expression of endoinulinase was dependent on the culture temperature and shaking. The highest expression of endoinulinase was observed at $30^{\circ}C$, and 150rpm.

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Studies on Cellulase Induction in Myriococcum albomyces (Myriococcum albomyces에 있어서 Cellulase 유도생성에 관한 연구)

  • Chung, Dong-Hyo
    • Korean Journal of Food Science and Technology
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    • v.3 no.1
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    • pp.1-5
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    • 1971
  • 1. Formation of cellulase in Myriococcum albomyces was investigated using shaking culture with addition of CMC or Avicel as an inducer to 5% wheat bran medium. 2. Three different types of cellulase fraction I, fraction II and fraction III in the culture filtrate were purified by elution column chromatography on a DEAE-Sephadex A-25. 3. By the addition of CMC as an inducer, CMCase activity was stronger than that of Avicelase. On the other hand, the addition of Avicel increased Avicelase activity.

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Detection of Lignans from Transformed Root Cultures of Schisandra chinensis Baillon (오미자의 형질전환된 근으로부터 리그난 화합물의 검출)

  • Hwang, Sung-Jin;Pyo, Byoung-Sik;Hwang, Baik
    • Korean Journal of Medicinal Crop Science
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    • v.12 no.6
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    • pp.448-453
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    • 2004
  • Transformed roots of Schisandra chinensis were obtained following co-cultivation of in vitro cultivated plantlet segments with Agrobaterium rhizogens ATCC15834. This root was examined for its growth and gomisin J contents under various culture conditions. Among the six basal culture media tested, WPM (Lloyd & McCown, 1980) medium supplemented with 5% sucrose was the best roots growth 6.2 (g D.W/flask) and gomisin J accumulation 1.56 $(X10^{-3}\;ug/g\;D.W)$. Initial inoculum size correlated with the yield of biomass while gomisin J contents was not affect. Gomisin J production was influenced by the initial sucrose concentration and the highest production yield was achieved at the concentration of 7%. The optimal shaking speeds for roots growth and gomisin J production was 120 and 140 rpm, respectively.

Isolation Identification and Physiological Characteristics of Some Sulfur-Reducing Microbes (수종 유황환원균주의 분리, 동정 및 그의 생리적 특성에 대하여)

  • 이민재;오명수
    • Korean Journal of Microbiology
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    • v.10 no.4
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    • pp.175-190
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    • 1972
  • This work was designed to illustrate physiological effects on the elimination of sulfur and its compounds in petroleum using sulfur-reducing bacteria. Deulfurizing bacteria were collected from sewage and soil at several areas in Soul and Ulsan, Korea. Seven supecies of sulfur-reducing microbes isolated were identified as : Pseudomonas marginata, Ps.effusa, Ps. putrefaciens, Ps.pseudcmcnelli, Ps. xanthochlora, Ps.bowlesiae, and Ps.aeruginosa. And some experiments were performed to define the growing characteristics of the Pseudomonads and the results obtained are as follows : 1) Shaking culture method was more effective for the growth of the cells than stagnant culture. 2) Beef peptone medium was better for the growth than other media. 3) Cuprous chloride of 50 ppm and cupper sulfate of 300 ppm treated, respectively, in the medium were effective for the growth. 4) Benzene of toluene of 5,000 ppm and petroleum ether of 50,000 ppm did not show remarkable inhibitory effects on the growth.

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Formation of protoplasts from pyricularia oryzae (벼 도열병균, pyricularia oryzae의 원형질체 형성)

  • 이용환;정후섭
    • Korean Journal of Microbiology
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    • v.23 no.3
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    • pp.209-214
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    • 1985
  • The optimum conditions of protoplasts formation from Pyricularia oryzae were investigated with lytic enzymes and osmotic stabilizers. The mycelia were begun to refease the protoplasts in response to the complex enzyme solution after 30-60 minutes and reached to maximum after 2-3hrs. Among the lytic enzymes tested, the mixture solution containing ${\beta}-Glucuronidase$(0.01 ml/ml), Cellulase ONOZUKA-RS(20mg/ml), Driselase (10mg/ml), and Macerozyme R-10 (10mg/ml) resulted in the highest rate of protoplasts releasing of Pyricularia oryzae. The best stabilizer was 0.6M KCl at pH 7.0. Shen the mycelia were digested with enzyme mixture, the stationary culture was better than shaking culture for higher protoplast formation.

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The Bioconversion of Red Ginseng Ethanol Extract into Compound K by Saccharomyces cerevisiae HJ-014

  • Choi, Hak Joo;Kim, Eun A;Kim, Dong Hee;Shin, Kwang-Soo
    • Mycobiology
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    • v.42 no.3
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    • pp.256-261
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    • 2014
  • A ${\beta}$-glucosidase producing yeast strain was isolated from Korean traditional rice wine. Based on the sequence of the YCL008c gene and analysis of the fatty acid composition, the isolate was identified as Saccharomyces cerevisiae strain HJ-014. S. cerevisiae HJ-014 produced ginsenoside Rd, $F_2$, and compound K from the ethanol extract of red ginseng. The production was increased by shaking culture, where the bioconversion efficiency was increased 2-fold compared to standing culture. The production of ginsenoside $F_2$ and compound K was time-dependent and thought to proceed by the transformation pathway of: red ginseng extract ${\rightarrow}Rd{\rightarrow}F_2{\rightarrow}$ compound K. The optimum incubation time and concentration of red ginseng extract for the production of compound K was 96 hr and 4.5% (w/v), respectively.