• Title/Summary/Keyword: Cellulomonas

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Factors affecting the protoplast formation and regeneration of Bacillus pumilus and Cellulomonas fimi for intergeneric protoplast fusion

  • Kim, D.M.;Lee, K.H.
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 1986.12a
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    • pp.527.3-527
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    • 1986
  • Several factors predicted to affect the protoplast formation and regeneration were investigated. The optimum lysozyme, casamino acid and PVP concentration were 0.5 (mg/$m\ell$), 0.1 (%) and 1.5(%). In B. pumilus, Penicillin-G treatment concentration was 0.3 (U/$m\ell$) and optimum treatment period was transit log. phase. And in the case of Celm. fimi, 0.3 (U/$m\ell$) and initial log. phase. Osmotic stabilizer and di-cation for OSM medium of B.pumilus and Gelm .fimi were 25mM CaCl2, 0.5M sodium sucinate and 50mM MgCl$_2$, 100mM CaCl$_2$, 0.4M sodium succinate. The regeneration frequency of B.pumilus and Celm. fimi were 14.6(%) and 6.9(%).

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Celluomonas sp. AP-7이 생산하는 Ascorbic Acid Phosphorylating Enzyme의 정제 및 특성

  • 이상협;최현일;방원기
    • Microbiology and Biotechnology Letters
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    • v.25 no.3
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    • pp.271-276
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    • 1997
  • An ascorbic acid phosphorylating enzyme, which catalyzes the formation of ascorbic acid-2-phosphate from ascorbic acid and pyrophosphate, was purified 32.7-folds to homogeneity from a cell-free extract of Cellulomonas sp. AP-7. The combination of DEAE- Sephacel ion exchange chromatography and Sephacryl S-200 get filtration was used for their purification. The molecular weight of the native protein was estimated to be 96.lkDa on high performance gel filtration chromatography. The SDS-PAGE analysis indicated that the protein consisted of four identical subunits of 24.6 kDa. The purified enzyme showed the optimal tempeature of 40$\circ$C and optimal pH of 4.5. The Km for ascorbic acid and pyrophosphate were 119 mM and 11.9 mM, respectively. The addition of 5,5'-dithiobis-(2-nitrobenzoic acid) into the reaction mixture resulted in the reduction of the enzyme activity at 51%. The enzyme also had a phosphatase activity at weakly acidic pH and the Km for ascorbic acid-2-phosphate in phosphatase activity was 7.9 mM.

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Screening of Methotrexate-Resistant Strains with High Thymidylate Synthase Activity (티미딜산 생성효소 활성이 높은 메토트렉세이트-내성 균주의 검색)

  • Kwak, In-Young;Lee, Jong-Soo
    • YAKHAK HOEJI
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    • v.36 no.4
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    • pp.345-349
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    • 1992
  • Thymidylate synthase activity from extracts of various methotrexate-resistant strains was measured by spectrophotometric assay. Methotrexate-resistant strains of Lactobacillus, Pseudomonas sp., Micrococcus sp. HS-1, Klebsillela pneumonae, Cellulomonas fimi and Serratia marcescens elevated thymidylate synthase levels, especially, Pseudomonas sp. KL-9 resistant to $10^{-9}M$ methotrexate have a 156-fold increase in thymidylate synthase, which suggests that Pseudomonas sp. is a convenient source of thymidylate synthase. Several methotrexate strains of yeast were tested, however, their enzyme activity was generally lower than that of bacteria tested.

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Characterization of TNP-cellulose as Substrate for Cellulase Assay (TNP-cellulose의 섬유소 분해효소 활성도 측정을 위한 기질로서의 특성)

  • Maeng, Jeong-Seob;Nam, Yoon-Kyu;Choi, Woo-Young
    • Korean Journal of Agricultural Science
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    • v.21 no.2
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    • pp.142-147
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    • 1994
  • Characteristics of TNP-cellulose which prepared from carboxymethyl cellulose powder, CM32, as substrate for cellulase activity assay were investigated. Enzymatic hydrolysis of TNP-cellulose occured on the cellulose moiety but not on amide bonds, following Michaelis-Menten kinetics. Three cellulase preparations from Trichoderma viride, Aspergillus niger, and Cellulomonas sp. were tested for their pH and temperature dependences and compared with the method determining the increase in reducing power. The enzyme activity was found to have the same temperature range in both methods, however the pH range was broadened in the case of using TNP-cellulose as substrate. The colorimetric method for cellulase assay using TNP-cellulose as substrate was compared with the other methods: one based on determination of the increase in reducing power; and the other based on determining the decrease in viscosity of Na-CM-cellulose solution. The activities measured by the colorimetric method showed a linear correlation with the enzyme concentration of certain range in all three enzymes tested, and the activity values were proportional to those obtained from the other methods. Depending on the enzyme, however, the activity values from this method were not always in proportion to those from the viscometric method. suggesting that this method was not specific for determination of the endo-type cellulase.

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Effect of Plant-growth-promoting Bacteria Inoculation on the Growth and Yield of Cucumber(Cucumis sativa L.) (식물생육촉진 세균이 오이 생육 및 수량에 미치는 영향)

  • Lee, Young-Han;Cho, Woo-Suk;Kim, Jong-Gyun;Lee, Han-Saeng;Park, Sang-Ryeol;Yun, Han-Dae
    • Korean Journal of Soil Science and Fertilizer
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    • v.30 no.2
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    • pp.196-199
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    • 1997
  • We studied the effect of inoculation of microorganisms known to produce plant growth promoting substances, on the growth and yield of cucumber(Cucumis sativa L.), through a field experiment. The microorganisms used were isolated from the forest soil and consisted of Micrococus sp., Baccilus sustilis, Enterobacter agglomerans, Baccilus megaterium, Pseudomonas putida, Cellulomonas sp. and Staphylococus xyposus. Fotr the multiplication, microorganisms were cultured in liquid media of Pseudomonas P and Sabouraud dextrose. Inoculation of microorganisms was done by spraying the culture media after the culture of them to soil and cucumber plants, three times during the growth of cucumber at the rate of 10l/ha. The inoculation of microorganisms tended to promote the growth of cucumber plant and increase the yield of it. No sign of significant improvement of soil chemical and physical properties were observed after the harvest of crop. The population of bacteria and actinomycetes tended to be higher in the inoculated plots than in not inoculated plots, while opposite was the case in the population of fungi.

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Identification of Enteric Bacteria from Nephila clavata (한국산 무당거미(Nephila clavata)에서 분리한 장내 세균의 동정)

  • 문은영;오현우;맹필재;배경숙
    • Korean Journal of Microbiology
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    • v.37 no.1
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    • pp.1-8
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    • 2001
  • Spiders are carnivores that prey upon insects and other small arthropods through digestion of food outside the body. Although spider poison may contain proteolytic enzymes, these are thought to play an insignificant role in actual digestion. The source of active proteolytic enzymes can be either the digestive tract cells of spider, or natural microbial flora in the digestive tract of spider. In this study, digestive tracts from the spider, Nephila clavata, were screened for bacteria that have protease or lipase activity. A total of $10^3-10^5$ CFU was recovered from a spider and more than 90% of them showed protease and lipase activity respectively. Of the microbial isolates, 63.3% showed protease or lipase activity, and 50% of these showed both protease and lipase activity. Some of the isolates were characterized using a battery of chemical, phenotypic and genotypic methods. Eleven Gram negative bacteriaa (Acinetobacter calcoaceticus, A. haemolyticus, Alcaligenes faecalis, Cedecea davisae, C. neteri, Klebsiella pneumoniae, Proteus vulgaris, Pseudomonas fluorescens, Serratia marcescens, Stenotrophomonas maltophilia, Suttonella indologenes) and 11 Gram positive bacteria (Bacillus cereus, B. coagulans, B. pasteurii, B. thuringiensis, Cellulomonas flavigena, Corynebacterium martruchotii, Enterococcus durans, E. faecalis, Micrococcus luteus, Staphylococcus hominis, S. sciuri) were identified.

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Studies on Microbial Utilization of Agricultural Wastes (Part IV) Cellulosic Waste Materials as Substrate on the Production of Cellulosic Single Cell Protein. (농산폐자원의 미생물학적 이용에 관한 연구 (제육보) 섬유소단세포단백 생산에서의 천연기질의 이용성)

  • Bae, Moo;Ko, Young-Hee
    • Microbiology and Biotechnology Letters
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    • v.5 no.1
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    • pp.18-23
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    • 1977
  • Experiments were carried out to pursue the availability and the feasibility of utilizable cellulosic materials as substrate for the production of cellulosic single cell protein. The resuluts were obtained as fellows. 1. Effects of carbolydrates as a sole carton source on the growth of Cellulomonas flavigena KIST 321 were examined. The result showed that cellulose and xylose would be most utlizable for cell mass production. 2. Alkaline treated waste papers and clothes resulted in good growth of the organism than intact ones did. However the waste papers as substrate of cellulosic fermentation were not digestible, even if the meterial was treated with alkalies. 3. Rice straw, rape straw and panic grass appeared to be good substrates for the cell mass production. 4. Leaves were proved to be a good substrate for the cell mass production, but wood sawdust was hardly digested by merely alkaline treatment. 5. When cellulosic wastes as the substrate were examined into the concentration of alkaline solution, the result suggested that the best productivity of cell mass from cellulosic materials was obtained on treatment with 0.8∼1.0% NaOH solution. 6. The productivity of cell mass was increased by washing out with water after alkaline treatment of newspaper, pine sawdust, lime sawdust and pine leaf.

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Thermostable ${\beta}$-Glycosidase-CBD Fusion Protein for Biochemical Analysis of Cotton Scouring Efficiency

  • Ha, Jae-Seok;Lee, Young-Mi;Choi, Su-Lim;Song, Jae-Jun;Shin, Chul-Soo;Kim, Ju-Hea;Lee, Seung-Goo
    • Journal of Microbiology and Biotechnology
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    • v.18 no.3
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    • pp.443-448
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    • 2008
  • Multidomain proteins for the biochemical analysis of the scouring efficiency of cotton fabrics were constructed by the fusion of a reporter moiety in the N-terminal and the cellulose binding domain (CBD) in the C-terminal. Based on the specific binding of the CBD of Cellulomonas fimi exoglucanase (Cex) to crystalline cellulose (Avicel), the reporter protein is guided to the cellulose fibers that are increasingly exposed as the scouring process proceeds. Among the tested reporter proteins, a thermostable ${\beta}$-glycosidase (BglA) from Thermus caldophilus was found to be most appropriate, showing a higher applicability and stability than GFP, DsRed2, or a tetrameric ${\beta}$-glycosidase (GUS) from Escherichia coli, which were precipitated more seriously during the expression and purification steps. When cotton fabrics with different scouring levels were treated with the BglA-CBD and incubated with X-Gal as the chromogenic substrate, an indigo color became visible within 2 h, and the color depth changed according to the conditions and extent of the scouring.

Twenty-five unrecorded bacterial species of the Republic of Korea belonging to the phylum Actinomycetota discovered during surveys in 2021

  • Inhyup Kim;Wan-Taek Im;Kiseong Joh;Myung Kyum Kim;Jung-Hoon Yoon;Wonyong Kim;Taegun Seo
    • Journal of Species Research
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    • v.12 no.3
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    • pp.229-239
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    • 2023
  • We isolated and identified 25 unrecorded bacterial species belonging to the phylum Actinomycetota found in the Republic of Korea. Sequence comparison of 16S rRNA was performed using the NCBI BLAST and EzBioCloud database to identify 25 species, which had a 16S rRNA gene sequence similarity of >98.8% and were allocated as unrecorded species in the Republic of Korea. Among the 25 unrecorded bacterial strains, Streptomyces was the most common with nine species, followed by Leifsonia with two species. Isoptericola, Nocardioides, Dermacoccus, Sinomonas, Patulibacter, Marmoricola, Allobranchiibius, Aldersonia, Actinokineospora, Agromyces, Aeromicrobium, Cellulomonas, and Gordonia with one species each were also found. Twenty-five unrecorded species were excavated in various environments, such as tidal flats, ferns, soil, pine cones, moss, mud, wetlands, and plants. These isolates were characterized on the basis of their phylogenetic, biochemical properties, and morphological data, and species descriptions were provided.

A report on 29 unrecorded bacterial species isolated from the Nakdonggang River, Republic of Korea

  • Ahyoung Choi;Ja Young Cho;Soo-Yeong Lee;Ji Young Jung;Kiwoon Baek;Seoni Hwang;Eui-Jin Kim;Jaeduk Goh
    • Korean Journal of Environmental Biology
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    • v.42 no.2
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    • pp.143-157
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    • 2024
  • As part of the research program "Freshwater Prokaryotic Organisms Research and Discovery," freshwater samples were collected from the Nakdonggang River. After plating the samples on several culture media and incubating aerobically, approximately 900 bacterial strains were isolated and identified using 16S rRNA gene sequences. Among the bacterial isolates showing higher than 98.7% 16S rRNA gene sequence similarity with those of already confirmed bacterial species previously unreported in Korea, 29 strains were selected. These strains were phylogenetically diverse and belonged to 3 phyla, 6 classes, 13 orders, and 21 genera. At the genus level, these previously unreported species were found to be affiliated with Novosphingobium, Sphingomonas, Polymorphobacter, Croceibacterium, Devosia, Endobacterium, Agaricicola, Bradyrhizobium, Paracoccus, and Pseudotabrizicola of the class Alphaproteobacteria; Undibacterium, Azonexus, and Dechloromonas of the class Betaproteobacteria; Acinetobacter and Budvicia of the class Gammaproteobacteria; Streptomyces, Nocardioides, Mycobacterium, and Cellulomonas of the phylum Actinomycetota; Flavobacterium and Pedobacter of the phylum Bacteroidota. These species were further characterized by examining their Gram reaction, colony and cell morphologies, biochemical properties, and phylogenetic positions. Detailed descriptions of these 29 previously unreported species are provided.