• Title/Summary/Keyword: substrate culture

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Microbial Conversion of Woody Waste into Sugars and Feedstuff (II) - Production of Cellulolytic Enzymes from Aspergillus fumigatus and Saccharification of Popla Wood (미생물(微生物)에 의한 목질자원(木質資源)의 당화(糖化) 및 사료화(飼料化)에 관(關)한 연구(硏究) (II) - Aspergillus fumigatus KC-1으로부터 섬유소 분해 효소의 생산 및 현사시나무의 효소가수분해)

  • Chung, Ki-Chul;Huh, Jeong-Weon;Myung, Kyu-Ho;Kim, Yoon-Soo
    • Journal of the Korean Wood Science and Technology
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    • v.15 no.4
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    • pp.18-25
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    • 1987
  • The cellulolytic activities of Aspergillus fumigatus KC-1 was investigated, which showed the most active producer of cellulase among the 256 strains of cellulose-decomposing microorganisms screened in our laboratory. All the examined cellulolytic activities (filter paper-, Avicel-, cotton-, CMC-, salicin- and xylansaccharifying activity) in a culture of A. fumigatus KC-1 grown on 1% popular sawdust pretreated with peroxide alkaline reached a maximum within 4-5 days. The optimum pH and temperature for the enzymatic activity was found to be pH 4.5 and $60^{\circ}C$ respectively. The sawdust of poplar wood delignified with 1% NaOH and 20% peracetic acid succesively recorded the highest hydrolysis rate in the tests of enzymatic saccharification. The major end product of hydrolysis of poplar wood with the cellulolytic enzymes obtained from A. fumigatus KC-1 was glucose with small amount of cellobiose and xylose. It can be concluded from these results that A. fumigatus KC-1 is an advantagous source of a cellulase that is capable of hydrolyzing cellulose to glucose rapidly. The influence of degree of delignification, substrate size and its concentration on the rate of hydrolysis of poplar wood was also discussed.

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Effect of seeding using an avidin-biotin binding system on the attachment of periodontal ligament fibroblasts to nanohydroxyapatite scaffolds: three-dimensional culture

  • Jang, Yong-Ju;Jung, Im-Hee;Park, Jung-Chul;Jung, Ui-Won;Kim, Chang-Sung;Lee, Yong-Keun;Kim, Chong-Kwan;Choi, Seong-Ho
    • Journal of Periodontal and Implant Science
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    • v.41 no.2
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    • pp.73-78
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    • 2011
  • Purpose: For periodontal tissue engineering, it is a primary requisite and a challenge to select the optimum types of cells, properties of scaffold, and growth factor combination to reconstruct a specific tissue in its natural form and with the appropriate function. Owing to fundamental disadvantages associated with using a two-dimensional substrate, several methods of seeding cells into three-dimensional scaffolds have been reported and the authors have asserted its usefulness and effectiveness. In this study, we explore the cell attachment of periodontal ligament fibroblasts on nanohydroxyapatite (n-HA) scaffold using avidin biotin binding system (ABBS). Methods: Human periodontal ligament fibroblasts were isolated from the health tooth extracted for the purpose of orthodontic procedure. HA nanoparticles were prepared and $Ca(NO_3)_2-_4H_2O$ and $(OC_2H_5)_3P$ were selected as precursors of HA sol. The final scaffold was 8 mm in diameter and 3 mm in height disk with porosity value of 81.55%. $1{\times}10^5$ periodontal ligament fibroblasts were applied to each scaffold. The cells were seeded into scaffolds by static, agitating and ABBS seeding method. Results: The number of periodontal ligament fibroblasts attached was greater for ABBS seeding method than for static or agitating method (P<0.05). No meaningful difference has been observed among seeding methods with scanning electron microscopy images. However, increased strength of cell attachment of ABBS could be deduced from the high affinity between avidin and biotin ($Kd=10^{-15}\;M$). Conclusions: The high-affinity ABBS enhances the ability of periodontal ligament fibroblasts to attach to three-dimensionally constructed n-HA scaffold.

Studies on the cellulase properties of Aspergillus clavatus from the Cellulose-Cultural Properties (섬유질 문화재로부터 분리된 Aspergillus clavatus의 섬유소분해호소에 관한 연구)

  • Chung, Hee-Jin;Han, Sung-Hee;Ahn, Hee-Kyun;Min, Kyung-Hee
    • The Korean Journal of Mycology
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    • v.15 no.1
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    • pp.29-37
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    • 1987
  • Cellulolytic mocrooganisms were isolated from the cellulose-cultural properties. Among them, Aspergillus clavatus with the highest cellulase activity was identified by the morphological characteristics and it's enzyme activities were compared on the various cultural conditions. It was found that the induction of carboxymethylcellulase(CMCase), avicelase and salicinase in CMC liquid media showed the highest enzyme acitivity on five day's cultivation at $30^{\circ}C$ and thereafter their activities were gradually decreased with time. After crude extracellular enzymes precipitated with the 70% saturated ammonium sulfate solution were dialyzed with 20 mM acetate buffer pH 6.0, each crude enzyme was examined. The optimal activities of CMCase and avicelase were both found to be at $50^{\circ}C$ and pH 6.0. Their thermal stability was appeared at the $50^{\circ}C$. CMCase and avicelase had the maxima activities with 1.5% and 2.2% substrate concentration, respectively. The concentration of 5 mM $Mg^{++}$ or $Ca^{++}$ was found to have a maximum cellulase activity and its activity was inhibited with more than 5 mM $Cu^{++}$ and $Zn^{++}$ concentration. Cellulase activity was also inhibited sensitively by the inhibitors such as 2-mercaptoethanol, iodine and sodium azide.

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Production of L-Lactic Acid from Soluble Starch by Enterococcus sp. JA-27. (Enterococcus sp. JA-27에 의한 가용성 전분으로부터 L형 젖산의 생산)

  • 김경아;김미경;장경린;전홍기
    • Microbiology and Biotechnology Letters
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    • v.31 no.3
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    • pp.250-256
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    • 2003
  • Lactic acid bacteria with amylolytic and acid producing activities can ferment starch directly to lactic acid thereby producing a monomer for the production of biodegradable poly lactic acid (PLA). In this study, the strain producing L-lactic acid from soluble starch was isolated from Nuruk. The isolated strain was identified as Enterococcus sp. through its morphological, cultural, biochemical characteristics as well as the 16S rDNA sequence analysis, and named Enterococcus sp. JA-27. Enterococcus sp. JA-27 produced exclusively L-lactic acid from soluble starch as a carbon source. The optimal conditions for the maximum production of L-lactic acid from Enterococcus sp. JA-27 were 30 C, pH 8, 1.5 % soluble starch as a substrate and 3.5 % tryptone as a nitrogen source, 0.1 % $K_2$$HPO_4$, 0.04 % $MgSO_4$. $7H_2$O, 0.014 % $MnSO_4$$.$4$H_2O$, 0.004% $FeSO_4$$.$$7H_2$O. Batch and fed batch culture were carried out and the former was more effective. L-Lactic acid production in the optimum medium was significantly increased in a 7 L jar fermenter, where the maximum L-lactic acid concentration was 3 g/L. For the purification of lactic acid in fermented broth, two stage ionexchange column chromatographies were employed and finally identified by HPLC.

Development of Passive Nutrient Supplying System and Its Effects on the Growth of Muskmelon (Cucumis melo L.) (수동방식 양액공급 시스템 개발과 멜론 재배 효과)

  • Nam, Sang-Sik;Oh, Yong-Bee;Kim, Yong-Bum;Choi, In-Hu
    • Horticultural Science & Technology
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    • v.19 no.3
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    • pp.338-341
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    • 2001
  • A new passive nutrient supplying system (PNS) was designed. The experiment was conducted to compare PNS with automatic hydroponic system (AHS) by investigating the growth characteristics of muskmelon (Cucumis melo L.) in spring and summer. No significant differences in growth characteristics of leaf area and shoot dry weight were observed between PNS and AHS. However, better netting of melon fruit was shown in PNS. Sugar content of melon fruit was also $1.0-2.0Brix^{\circ}$ higher in PNS than in AHS. The suitable substrate for melon culture with PNS was the mixture of perlite 70% and rice hull 30%. These results suggested that new PNS could be introduced to growers without any loss of fruit yield and quality of muskmelon.

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Conversion of Ginseng Saponin with the Enzyme Produced by Rhizopus sp. (Part 1) Confirmation of Conversion of Ginsenoside- Rb$_1$to Ginsenoside-Rd (Rhizopus sp.가 생산하는 효소에 의한 인삼 Saponin의 전환 (제1보) Ginsenoside-Rb$_1$에서 Ginsenoside-Rd로의 전환확인)

  • 김상달;서정훈
    • Microbiology and Biotechnology Letters
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    • v.10 no.4
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    • pp.267-273
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    • 1982
  • Among 12 kinds of ginsenosides in ginseng saponin, ginsenoside-Rb$_1$was contained the most abundantly. But ginsenoside-Rd which is similar to ginsenoside-Rb$_1$in structure, was known to be superior to ginsenoside-Rb$_1$pharmaceutically. In order to convert ginsenoside-Rb$_1$into ginsenoside-Rd by microbial enzyme treatment, a Rhizopus sp. was selected among various strais of molds found in rotten ginseng roots. Enzyme was prepared from the extract of wheat bran koji culture by ammonium sulfate precipitation (1.0 sat'd) and succeeding ammonium sulfate fractionation method (0.6-0.9 sat'd). For the purpose of use as substrate, saponins were purified by the several purification steps from alcohol extract of red ginseng roots. We obtained the total saponin which was composed of 36.5% of ginsenoside Rb$_1$, 12.2% of ginsenoside-Rd and other ginsenosides. For increase of ginsenoside-Rb$_1$ component ratio, we also obtained further purified ginsenoside-Rb group saponin containing 54.5% of ginsenoside-Rb$_1$, 1.1% of ginsenoside- Rd and other ginsenosides from purified the total saponin. In the enzymatic reaction system including the total saponin or the ginsenoside-Rb group saponin, we confirmed the specific conversion of ginsenoside-Rb$_1$to ginsenoside-Rd proportionally and no change of any other ginsenoside patterns by thin layer chromatography and high performance liquid chromatography.

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Isolation of an Agarolytic Bacteria, Cellvibrio mixtus SC-22 and The Enzymatic Properties (한천분해세균 Cellvibrio mixtus SC-22의 분리 및 효소적 특성)

  • Cha, Jeong-Ah;Kim, Yoo-Jin;Seo, Yung-Bum;Yoon, Min-Ho
    • Journal of Applied Biological Chemistry
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    • v.52 no.4
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    • pp.157-162
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    • 2009
  • An agar-liquefying bacteria (SC-22), which produces a diffusible agarase that caused agar softening around the colony was isolated from Daecheong lake in Korea. Chemotaxanomic and phylogenetic analyses based on 16S rRNA gene sequences revealed the strain was classified as Cellvibrio mixtus SC-22. The isolate SC-22 showed maximal extracellular agarase activity with 58.5 U/mL after 48 h cultivation in the presence of 0.2% agar. It was observed that the isolate produced two kinds of extracellular and three kinds of intracellular isoenzymes. The major agarase was purified from the culture filtrate of agarolytic bacteria by ammonium sulfate precipitation, anion exchange and gel filtration column chromatographic methods. The molecular mass of the purified enzyme was estimated to be 25 kDa by SDS-PAGE. The optimum pH and temperature of the purified enzyme were pH 7.0 and $50^{\circ}C$, respectively. The agarase activity was activated by $Fe^{2+}$, $Na^+$ and $Ca^{2+}$ ions while it was inhibited by $Hg^{2+}$, $Mn^{2+}$ and $Cu^{2+}$ at 1 mM concentration. The predominant hydrolysis product of agarose by the enzyme was galactose and disaccharide on TLC, indicating the cleavage of $\beta$-1,4 linkage in a random manner. The enzyme showed high substrate specificity for only agar and agarose among various polysaccharides.

Culture of Potted $Viola{\times}wittrockiana$ 'Majestic GT Scarlet Shadow' in Various Mixtures of Recycled Perlite and CGF in a Mat Subirrigation System (재사용한 펄라이트 및 다공성 CGF의 혼합배지와 매트저면관수를 이용한 분화$Viola{\times}wittrockiana$ 'Majestic GT Scarlet Shadow'의 재배)

  • Kim Gyeong-Hee;Lee Kang-Mo;Jeong Byoung-Ryong
    • Journal of Bio-Environment Control
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    • v.15 no.3
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    • pp.239-243
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    • 2006
  • This research compared growth and flowering of potted $Viola{\times}wittrockiana$ 'Majestic GT Scarlet Shadow' in various mixtures of recycled perlite and cellular glass foam (CGF). The crop was cultured in a mat subirrigation system. Numbers of leaves, opened flowers, flower buds, and branches, fresh and dry weights of shoot, leaf area, and total chlorophyll concentration were the greatest in plants grown in the medium of used perlite + granular rockwool+ peatmoss (25:50:25, v/v/v). Plant height and length of the longest root were the highest and longest, respectively, in plants grown in the medium of CGF+peatmoss (25:75, v/v). The medium of CGF+peatmoss (25:75, v/v) was better than the medium of used perlite+peatmoss (25:75, v/v) in plant growth. The medium of CGF + granular rockwool (25:75, v/v) was poorer than the medium of recycled perlite + granular rockwool (25:75, v/v).

Effect of Exogenous Fibrolytic Enzyme Application on the Microbial Attachment and Digestion of Barley Straw In vitro

  • Wang, Y.;Ramirez-Bribiesca, J.E.;Yanke, L.J.;Tsang, A.;McAllister, T.A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.1
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    • pp.66-74
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    • 2012
  • The effects of exogenous fibrolytic enzymes (EFE; a mixture of two preparations from Trichoderma spp., with predominant xylanase and ${\beta}$-glucanase activities, respectively) on colonization and digestion of ground barley straw and alfalfa hay by Fibrobacter succinogenes S85 and Ruminococcus flavefaciens FD1 were studied in vitro. The two levels (28 and 280 ${\mu}g$/ml) of EFE tested and both bacteria were effective at digesting NDF of hay and straw. With both substrates, more NDF hydrolysis (p<0.01) was achieved with EFE alone at 280 than at 28 ${\mu}g$/ml. A synergistic effect (p<0.01) of F. succinogenes S85 and EFE on straw digestion was observed at 28 but not 280 ${\mu}g$/ml of EFE. Strain R. flavefaciens FD1 digested more (p<0.01) hay and straw with higher EFE than with lower or no EFE, but the effect was additive rather than synergistic. Included in the incubation medium, EFE showed potential to improve fibre digestion by cellulolytic ruminal bacteria. In a second batch culture experiment using mixed rumen microbes, DM disappearance (DMD), gas production and incorporation of $^{15}N$ into particle-associated microbial N ($^{15}N$-PAMN) were higher (p<0.001) with ammoniated (5% w/w; AS) than with native (S) ground barley straw. Application of EFE to the straws increased (p<0.001) DMD and gas production at 4 and 12 h, but not at 48 h of the incubation. EFE applied onto S increased (p<0.01) $^{15}N$-PAMN at 4 h only, but EFE on AS increased (p<0.001) $^{15}N$-PAMN at all time points. Prehydrolysis increased (p<0.01) DMD from both S and AS at 4 and 12 h, but reduced (p<0.01) $^{15}N$-PAMN in the early stage (4 h) of the incubation, as compared to non-prehydrolyzed samples. Application of EFE to barley straw increased rumen bacterial colonization of the substrate, but excessive hydrolytic action of EFE prior to incubation decreased it.

Tetraspore Release and Growth of a Crustose Coralline Alga, Lithophyllum yessoense (Rhodophyta, Corallinaceae) (홍조류 무절석회조, 납작돌잎 (Lithophyllum yessoense) 사분포자체의 포자방출과 생장)

  • Hwang Eun Kyoung;Kim Eun Jin;Kim Hyung Geun;Sohn Chul Hyun
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.35 no.3
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    • pp.242-246
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    • 2002
  • Growth and tetraspore release pattern of lithophyllum yessoense (Rhodophyta, Corallinaceae) were investigated from March 2000 to July 2001. Pinkish tetraspores were 40.2 $\pm$ 0.4 $\mu$m in diameter. After release, tetraspores attached on substrate shortly. Culture conditions were five temperatures (5, 10, 15, 20, 25^{\circ}C), four irradiances (0, 20, 50, 100 $\mu$mol${\cdot}m^{-2}$${\cdot}s^{-1}$) and six salinities (0, 9, 17, 25, 34, 43 ppt). Maximum growth of gametophyte was occurred at $20^{\circ}C$, $20{\mu}mol\;m^{-2}$${\cdot}s^{-1}$, 16: 8h (L:D) and 34 put. Maximum relative growth rate was 0.1232 at $20^{\circ}C$. The amount of tetraspore release showed maximum at September as 266 cells per crust area ($cm^2$), and tetraspores did not release from January to March.