• Title/Summary/Keyword: Substrate culture

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Afforestation of a Brown Alga, Ecklonia cava Kjellman using a Biodegradable Polybutylene Succinate (생분해성 로프를 이용한 대형 갈조류 감태의 이식)

  • Baek, Jae-Min;Park, Seong-Wook;Hwang, Eun-Kyoung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.42 no.5
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    • pp.523-526
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    • 2009
  • Biodegradable polybutylene succinate (BPS) multifilament were designed to degrade upon disposal by the action of afforestration of Ecklonia cava. Matured thalli of E. cava were collected at Jeju for zoospore collection and the substrate for zoospores were BPS multifilament (12 mm, 500 Td/96). The materials were made as nets of $1\;m^2$ with a cross stripes of 10 cm. The unit biodegradable nets bearing germlings of E. cava was moved into Wando where the place is conducting intensive seaweed cultivation in Korea for 5 months of nursery culture until they grew to 10 cm in length after which the nets were transplanted into the sea bed at Jeju at a depth of 12 m and the algal growth was monitored from May 2007 to December 2008. This is the first instance of using the BPS materials for seaweed afforestation to avoid any environmental problems.

Spatial Heterogeneity of Bacteria: Evidence from Hot Composts by Culture-independent Analysis

  • Guo, Yan;Zhang, Jinliang;Deng, Changyan;Zhu, Nengwu
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.7
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    • pp.1045-1054
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    • 2012
  • The phylogenetic diversity of the bacteria in hot composting samples collected from three spatial locations was investigated by molecular tools in order to determine the influence of gradient effect on bacterial communities during the thermophilic phase of composting swine manure with rice straw. Total microbial DNA was extracted and bacterial near full-length 16S rRNA genes were subsequently amplified, cloned, restriction fragment length polymorphism-screened and sequenced. The superstratum sample had the highest microbial diversity among the three samples which was possibly related to the surrounding conditions of the sample resulting from the location. The results showed that the sequences related to Bacillus sp. were most common in the composts. In superstratum sample, 45 clones (33%) and 36 clones (27%) were affiliated with the Bacillus sp. and Clostridium sp., respectively; 74 clones (58%) were affiliated with the Clostridium sp. in the middle-level sample; 52 clones (40%) and 29 clones (23%) were affiliated with the Clostridium sp. and Bacillus sp. in substrate sample, respectively. It indicated that the microbial diversity and community in the samples were different for each sampling site, and different locations of the same pile often contained distinct and different microbial communities.

Production of a Biosurfactant Mannosylerythritol Lipid by Resting Cell of Candida sp. SY16. (Candida sp. SY16의 휴식세포를 이용한 생물계면활성제 Mannosylerythritol Lipid의 생산)

  • 김희식;전종운;최우영;오희목;이기형;권태종;윤병대
    • Microbiology and Biotechnology Letters
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    • v.30 no.2
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    • pp.167-171
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    • 2002
  • The resting cells of Candida sp. SY16 produced a large amount of mannosylerythritol lipid as a biosurfactant when incubated in the distilled water containing only the carbon source. The resting cells exhibited the highest production at 20 g cells per liter on the soybean oil of 75 g/1 as a sole substrate and pH 4∼5 in the shaking culture. Under the optimal conditions, the biosurfactant was extracellularly produced to 58 g/1 after 120 h in jar fermentor, and the yield became higher than that obtained by using the glowing cells of the strain in batch fermentation.

Purification and Characterization of Streptomyces griseus Trypsin Overexpressed in Streptomyces lividans

  • KOO, BON-JOON;KWANG HEE BAE;SI-MYONG BYUN;SOON-KWANG HONG
    • Journal of Microbiology and Biotechnology
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    • v.8 no.4
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    • pp.333-340
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    • 1998
  • Streptomyces griseus trypsin (SGT) is an extracellular proteinase produced by S. griseus. The sprT gene, which encodes premature SGT protein, was cloned into the plasmid pWHM3, a Streptomyces-E. coli shuttle vector. When the recombinant plasmid was introduced into Streptomyces lividans TK24, two proteins with molecular weights of 28 kDa and 42 kDa were detected. The 28-kDa protein was a SGT protein while the larger 42-kDa protein is thought to have been a premature form of the SGT protein. The SGT protein was purified to homogeneity via ammonium sulfate fractionation and many column chromatographies, including CM -sepharose chromatography, Mono-S chromatography, and Superose-12 chromatography, from the culture broth of S. lividans TK24 harboring the sprT gene. The N-terminal amino acid sequence, isoelectric points, and stabilities at various conditions of the SGT proteins purified from the Pronase and transformant were almost identical. The amount of the expressed SGT in S. lividans TK 24 was determined to be 5 times more than that of S. griseus based on the enzymatic activity against artificial substrate.

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Potential of Immobilized Whole-Cell Methylocella tundrae as a Biocatalyst for Methanol Production from Methane

  • Mardina, Primata;Li, Jinglin;Patel, Sanjay K.S.;Kim, In-Won;Lee, Jung-Kul;Selvaraj, Chandrabose
    • Journal of Microbiology and Biotechnology
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    • v.26 no.7
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    • pp.1234-1241
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    • 2016
  • Methanol is a versatile compound that can be biologically synthesized from methane (CH4) by methanotrophs using a low energy-consuming and environment-friendly process. Methylocella tundrae is a type II methanotroph that can utilize CH4 as a carbon and energy source. Methanol is produced in the first step of the metabolic pathway of methanotrophs and is further oxidized into formaldehyde. Several parameters must be optimized to achieve high methanol production. In this study, we optimized the production conditions and process parameters for methanol production. The optimum incubation time, substrate, pH, agitation rate, temperature, phosphate buffer and sodium formate concentration, and cell concentration were determined to be 24 h, 50% CH4, pH 7, 150 rpm, 30℃, 100 mM and 50 mM, and 18 mg/ml, respectively. The optimization of these parameters significantly improved methanol production from 0.66 to 5.18 mM. The use of alginate-encapsulated cells resulted in enhanced methanol production stability and reusability of cells after five cycles of reuse under batch culture conditions.

PRODUCTION OF HUMAN PROTEIN TIMP-2: A HIGHLY EFFECTIVE ANTI-AGING INGREDIENT

  • Schutz, R.;Imfeld, D.
    • Proceedings of the SCSK Conference
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    • 2003.09a
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    • pp.590-600
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    • 2003
  • The matrix metalloproteinases (MMPs) are a family of enzymes responsible for degrading connective tissue. MMPs catalyze the breakdown of collagen from the extracellular matrix, leading to wrinkle formation and accelerated skin aging. Furthermore, ultraviolet irradiation causes increased expression of certain MMPs. In the extracellular matrix turnover, MMPs are interacting with endogenous regulators named tissue inhibitors of metalloproteinases (TIMPs). Using peptide substrate assays, it has been demonstrated that TIMP-MMP complexes interact highly specifically with $K_{i}$ values of 10$^{-9}$ -10$^{-16}$ M. Therefore applications for TIMP as inhibitor of collagen degradation are suggested for cosmetic anti-aging products to prevent wrinkle formation and loss of elasticity. To date four TIMP proteins (TIMP-1, TIMP-2, TIMP-3 and TIMP-4) have been identified which show a high degree in sequence similarity. The production of human TIMP-2, a 194-residue nonglycosylated protein, was performed by fed-batch culture of Escherichia coli. TIMP-2 accumulated in the bacterial cells in an insoluble form as inclusion bodies. The inclusion bodies were solubilized and the protein refolded to yield the native TIMP-2 in the active form. The integrity of the protein was confirmed by mass analysis, Edman sequencing and gel shift experiments with authentic samples. The inhibitory activity of the refolded and purified TIMP-2 was demonstrated with MMP-1 and MMP-2 assays using synthetic fluorogenic peptide substrates.s.

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Characteristics of Microalgal Growth on Anaerobic Effluent of Animal Waste (축산분뇨 혐기성 처리수에서 미세조류의 성장특성)

  • Lim, Byung-Ran;Lee, Kisay;Noh, Seung You;Park, Ki Young
    • Journal of Korean Society on Water Environment
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    • v.24 no.3
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    • pp.306-310
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    • 2008
  • Characteristics of microalgal growth was investigated using anaerobic effluent from two-phase animal waste digestor as substrate. Batch experiments were carried out to investigate the effect of the initial nitrogen and phosphorus concentrations on growth of Microcystis aeruginosa, Chlorella sp. and Euglena gracilis. In 400 times diluted anaerobic effluent (TN 3 mg/L), single cell growth of the Euglena gracilis population increased twice without delay, although Chlorella sp. and Microcystis aerugenos take over 144 hours. Similar appearance with single cell growth was observed in mixed cultures. However, microalgae population did not increase under condition of 10 times diluted influent (TP 3 mg/L) in both pure and mixed cultures, which was affected by high organic and nitrogen concentration. Logistic growth model successfully fitted to determine biokinetic parameters such as ${\lambda}$: lag time, ${\mu}m$: maximal specific growth rate, A: asymptote of growth.

Biosynthesis of polyhydroxybutyrate and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) by bacillus thuringiensis R-510

  • Park, Sang-Kyu;Lee, Kang-Tae;Kim, Young-Baek;Rhee, Young-Ha
    • Journal of Microbiology
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    • v.35 no.2
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    • pp.127-133
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    • 1997
  • Biosynthesis of polyhydroxybutyrate and copolymer consisting of 3-hydroxybutyrate and 3-hydroxyvalerate [poly(3HB-co-3HV)] by Bacillus thuringiensis R-510 grown with glucose or with mixtures of glucose and propionate was investigated. n-Alkanoic acids other than propionate were not precursors of 3HV units. The fraction of 3HV unit in the copolymer increased from 0 to 84 mol% of 3HV. Polymer yield decreased as the fraction of propionate was increased but the molecular weight distribution was not affected by the composition of carbon substrate. The minimum melting temperature (around 65.deg.C) of poly (3HB-co-3HV) copolymers was observed for the polymer bearing approximately 35 mol% of 3HV. Polyhydroxyalkanoates production by this organism was not dependent on nutritional limitation, but remarkably influenced by dissolved oxygen concentration in the culture medium. Low level of dissolved oxygen concentration prevented spore formation in the cells and stimulated the synthesis of polyhydroxyalkanoate. The composition of poly (3HB-co-3HV) produced by B. thuringiensis R-510 lyhydroxyalkanoate. The composition of poly(3HB-co-3HV) propduced by B. thuringiensis R-510 varied according to the growth time. However, there was no evidence that polymers isolated from cells were mixtures of immiscible polymers.

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β-Glucosidase Formation In Cellulomonas sp. (Cellulomonas sp.의 β-글루코시다아제 생성)

  • Choi, Woo-Young
    • Korean Journal of Agricultural Science
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    • v.3 no.2
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    • pp.225-234
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    • 1976
  • To elucidate ${\beta}$-glucosidase formation on various carbon scurces by cellulolytic bact-eia, Cellulomonas sp. CS1-1, the strain was grown on Nutrient Yeast Broth, carboxymethyl cellulose, avicel and cellobiose using a Ouickfit FVIL fermentor operated in batch, and the growth characteristics on those substrates and ${\beta}$-glucosidase distribution of extra and intracellular enzyme components were studied. The results were: 1) ${\beta}$-glucosidase was always intracellular, and was formed under all growth conditions tested, ii) but levels of relative activities were higher when the culture was grown on cellobiose and on avicel, iii) the relative activities were always maximum during the growth phase of the organism irrespective of the substrate used.

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Purufication and Characterization of Extracellular Collagenase from Vibrio mimicus (Vibrio mimicus 가 생산하는 collagenase의 정제 및 특성)

  • 김용태;김세권
    • Journal of Life Science
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    • v.6 no.4
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    • pp.241-249
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    • 1996
  • A collagenase was isolated from the culture filtrate of Vibrio mimicus (ATCC 33658). The enzyme was purified to homogeneity by ammonium sulfate precipitation and DEAE-Sephadex A-50 chromatography, which an activity recovery of 22%. The molecular weight of the purified enzyme was estimated to be 42 kDa by SDS-polyacrylamide gel electrophoresis and gel filtration, indication a monomer structure. The optimum pH and temperature od the enzyme for insoluble collagen (Type I) were around 7.75 and 28$\circ$C, respectively. Some chelating agents and serine protease inhibitor inactivated the enzyme, but L-cysteine and histidine did not affect the activity. The amino acid composition indicated that the collagenase contained high amounts of amino acid residues of glycine and alanine. The K$_{m}$ and R$_{cat}$/K$_{m}$ values for the collagenase, using insoluble collagen (type I) as substrate, were 2.86 mg/ml and 972.28 U/mg-protein, respectively.

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