• Title/Summary/Keyword: starch isolation

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Isolation of Mutants Overproducing Amylase from Nuruk Fungi by NTG (NTG에 의한 Amylase활성이 높은 누룩사상균의 변이주의 분리)

  • 정혁준;김영숙;유대식
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.6
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    • pp.987-994
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    • 2000
  • Aspergillus coreanus NR 15-1, Asp. oryzae NR 15-3 and Asp. oryzae NR 2-5 isolated from traditional Korean nuruk were screened as parental strains producing starch hydrolyzing enzymes. They were mutagenized by N-methyl -N'-nitro-N-nitrosoguanidine (NTG) and mutants were isolated for analysis of various amylase activities and the ability of acid production. Among them, the mutants harboring high saccharogenic activity, dextrinogenic activity, and the ability of acid production were selected. Fifteen, six, and five strains of mutants were isolated from Asp. coreanus NR 15-1, Asp. oryzae NR 2-5, and Asp. oryzae NR 15-3, respectively followed by NTG mutagenesis. Among these mutants, thirteen strains were identified as auxotrophic mutants. \ulcorner (Arg. ̄) mutant from Asp. coreanus NR 15-1 showed high glucoamylase activity and total acid productivity. Z6 (Ade. ̄) mutant from Asp. oryzae NR 2-5 showed the highest $\alpha$-amylase activity, therefore \ulcorner and Z6 mutant were selected.

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Isolation of a Novel Freshwater Agarolytic Cellvibrio sp. KY-YJ-3 and Characterization of Its Extracellular ${\beta}$-Agarase

  • Rhee, Young-Joon;Han, Cho-Rong;Kim, Won-Chan;Jun, Do-Youn;Rhee, In-Ku;Kim, Young-Ho
    • Journal of Microbiology and Biotechnology
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    • v.20 no.10
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    • pp.1378-1385
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    • 2010
  • A novel agarolytic bacterium, KY-YJ-3, producing extracellular agarase, was isolated from the freshwater sediment of the Sincheon River in Daegu, Korea. On the basis of Gram-staining data, morphology, and phylogenetic analysis of the 16S rDNA sequence, the isolate was identified as Cellvibrio sp. By ammonium sulfate precipitation followed by Toyopearl QAE-550C, Toyopearl HW-55F, and MonoQ column chromatographies, the extracellular agarase in the culture fluid could be purified 120.2-fold with a yield of 8.1%. The specific activity of the purified agarase was 84.2 U/mg. The molecular mass of the purified agarase was 70 kDa as determined by dodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The optimal temperature and pH of the purified agarase were $35^{\circ}C$ and pH 7.0, respectively. The purified agarase failed to hydrolyze the other polysaccharide substrates, including carboxymethyl-cellulose, dextran, soluble starch, pectin, and polygalacturonic acid. Kinetic analysis of the agarose hydrolysis catalyzed by the purified agarase using thin-layer chromatography showed that the main products were neoagarobiose, neoagarotetraose, and neoagarohexaose. These results demonstrated that the newly isolated freshwater agarolytic bacterium KY-YJ-3 was a Cellvibrio sp., and could produce an extracellular ${\beta}$-agarase, which hydrolyzed agarose to yield neoagarobiose, neoagarotetraose, and neoagarohexaose as the main products.

Isolation of Bacillus subtilis CK-2 Hydrolysing Various Organic Materials (다양한 유기물을 분해하는 Bacillus subtilis CK-2의 분리)

  • Kim, Chul-Ho;Lee, Sang-Hyup
    • Journal of Life Science
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    • v.21 no.12
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    • pp.1716-1720
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    • 2011
  • A bacterium hydrolysing various organic materials including cellulose, protein, starch and lipid was isolated. The isolate was identified as Bacillus subtilis, and named Bacillus subtilis CK-2 in this paper. This bacterium showed optimal growth at $40\sim45^{\circ}C$, pH 6~9, and 0~3% of NaCl. B. subtilis CK-2 seemed to synthesis highly active autolysin. The hydrolytic enzymes produced by B. subtilis CK-2 were primary enzymes because extracellular enzyme activities varied similarly to the growth curve. The hydrolytic enzymes seemed to be stable at basic pH conditions. From these results, B. subtilis CK-2 was found to bea useful bacterial agent for composting, or for use in feed-production waste in agriculture, fishery, forest materials, livestock farming, and food.

Isolation and Identification of Burkholderia pyrrocinia CH-67 to Control Tomato Leaf Mold and Damping-off on Crisphead Lettuce and Tomato

  • Lee, Kwang-Youll;Kong, Hyun-Gi;Choi, Ki-Hyuck;Lee, Seon-Woo;Moon, Byung-Ju
    • The Plant Pathology Journal
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    • v.27 no.1
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    • pp.59-67
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    • 2011
  • A bacterial strain CH-67 which exhibits antagonism towards several plant pathogenic fungi such as Botrytis cinerea, Fulvia fulva, Rhizoctonia solani, Sclerotinia sclerotiorum, Colletotrichum sp. and Phytophthora sp. was isolated from forest soil by a chitin-baiting method. This strain was identified as Burkholderia cepacia complex (Bcc) and belonging to genomovar IX (Burkholderia pyrrocinia) by colony morphology, biochemical traits and molecular method like 16S rRNA and recA gene analysis. This strain was used to develop a bio-fungicide for the control of tomato leaf mold caused by Fulvia fulva. Various formulations of B. pyrrocinia CH-67 were prepared using fermentation cultures of the bacterium in rice oil medium. The result of pot experiments led to selection of the wettable powder formulation CH67-C containing modified starch as the best formulation for the control of tomato leaf mold. CH67-C, at 100-fold dilution, showed a control value of 85% against tomato leaf mold. Its disease control efficacy was not significantly different from that of the chemical fungicide triflumidazole. B. pyrrocinia CH-67 was also effective in controlling damping-off caused by Rhizoctonia solani PY-1 in crisphead lettuce and tomato plants. CH67-C formulation was recognized as a cell-free formulation since B. pyrrocinia CH-67 was all lethal during formulation process. This study provides an effective biocontrol formulation of biofungicide using B. pyrrocinia CH-67 to control tomato leaf mold and damping-off crisphead lettuce and tomato.

Isolation and Identification of the Antagonistic Microorganisms Against Streptococous spp. Causing Dental Caries in Korean Soy Sauce (한국재래간장으로부터 구강질환 방제균의 선발 및 동정)

  • 엄수정;이여진;김진락;이은탁;김상달
    • Journal of Life Science
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    • v.13 no.4
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    • pp.535-540
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    • 2003
  • The antagonistic microorganisms against Streptococcus sanguis, S. salivarius and S. mutans causing the dental caries of oral diseases were isolated from Korean traditional soy sauce. Twenty five strains were isolated by pairing culture, paper disc culture and dual culture methods. The isolate NG 06 strain was observed with various cultural and physiological test, and $Biolog^{(R)}$ Bacterial Identification System. The strain was identified as Bacillus racemilacticus. The isolate NG 16 strain was confirmed to Gram-positive, rods, endospore production, utilization of melibiose, casein hydrolysis and starch hydrolysis. Also the second strain NG 16 was identified as $\beta$. amyloliquefaciens.

Bioactivity of Metabolites from Actinomycetes Isolates from Red Sea, Egypt

  • Osman, Mohamed E.;El-nasr, Amany A. Abo;Hussein, Hagar M;Hamed, Moaz M
    • Microbiology and Biotechnology Letters
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    • v.50 no.2
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    • pp.255-269
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    • 2022
  • Actinomycetes isolated from marine habitats represent a promising source of bioactive substances. Here, we report on the isolation, identification, productivity enhancement and application of the bioactive compounds of Streptomyces qinglanensis H4. Eighteen marine actinomycetes were isolated and tested for resistance to seven bacterial diseases. Using 16S rRNA sequencing analysis (GenBank accession number MW563772), the most powerful isolate was identified as S. qinglanensis. Although the strain produced active compound(s) against a number of Gram-negative and Gram-positive bacteria, it failed to inhibit pathogenic fungi. The obtained inhibition zones were 22.0 ± 1.5, 20.0 ± 1, 16.0 ± 1, 12.0 ± 1, 22.0 ± 1 and 24.0 ± 1 mm against Bacillus subtilis ATCC 6633, Escherichia coli ATCC 19404, Enterococcus faecalis ATCC 29212, Pseudomonas aeruginosa ATCC 9027, Candida albicans ATCC 10231 and Staphylococcus aureus ATCC6538, respectively. To maximize bioactive compound synthesis, the Plackett-Burman design was used. The productivity increased up to 0.93-fold, when S. qinglanensis was grown in optimized medium composed of: (g/l) starch 30; KNO3 0.5; K2HPO4 0.25; MgSO4 0.25; FeSO4·7H2O, 0.01; sea water concentration (%) 100; pH 8.0, and an incubation period of 9 days. Moreover, the anticancer activity of S. qinglanensis was tested against two different cell lines: HepG2 and CACO. The inhibition activities were 42.96 and 57.14%, respectively. Our findings suggest that the marine S. qinglanensis strain, which grows well on tailored medium, might be a source of bioactive substances for healthcare companies.

DISTRIBUTION AND PHYSIOLOGICAL CHARACTERISTICS OF BACILLUS CEREUS IN RICE AND RICE PRODUCTS (미반류에 있어서 Bacillus cereus균의 분포와 생리적특성에 관한 연구)

  • LEE Myeong-Sook;CHANG Dong-Suck
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.13 no.4
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    • pp.163-171
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    • 1980
  • Recently, Bacillus has been identified as one of food poisoning bacteria especially in products of cereal foods in foreign countries. Therefore, the quantitative distribution of Bacillus cereus in market foods, its physiological characteristics, growth rate by temperature and heat resistance of its spore were examined. Thirty two samples of cooked rice, 20 samples of kimbab(cooked rice rolled with laver), 23 samples of rice cake, 13 samples of rice ana 13 samples of barley were collected from restaurents and food stores in Busan, Korea during the period from May to November in 1980. Forty samples of 101 samples submitted to the test appeared positive for Bacillus cereus showing abut $40\%$ in detection ratio. Detection ratio of Bacillus cereus was higher than $50\%$ in barley and rice, and about $30\%$ in rice products. Average Bacillus cereus content of in the samples was $2.6\times10^6/g$ in cooked rice, $2.3\times10^6/g$in kimbab, $4.9\times10^4/g$ in rice cake while that in rice and barley was about $10^3/g$. The result of biochemical tests of the bacterium was $100\%$ positive in catalase, egg yolk reaction, gelatin hydrolysis and glucose fermentation, $100\%$ negative in xylose, arabinose and mannitol oxidation, about $90\%$ positive in acetoin production, $80.0\%$ positive in nitrate reduction and citrate utilization and $55.0\%$ positive in starch hydrolysis test. Isolation ratio of Bacillus ceresus which showed haemolysis positive and starch hydrolysis negative results, was about $38\%$ in 40 strains examined. It is known that those strains has a close relation to food poisoning accident. Growth rate and generation time of Bacillus cereus isolated from the cooked rice were $0.34hr^{-1},\;2.02hr\;at\;20^{\circ}C,\;0.73hr^{-1},\;0.95hr\;at\;30^{\circ}C\;and\;0.49hr^{-1},\;1.44\;hr\;at\;40^{\circ}C$ respectively. Heat resistance value of Bacillus cereus spores suspended in phosphate buffer solution was $D_{90}=29.0min,\;D_{95}=8.7min,\;D_{98}=3.7\;min\;and\;D_{101}=2.3\;min(z=10.5)$.

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The Study on the Relationship between Changes of Rumen Microflora and Bloat in Jersey Cow (저지종 젖소의 반추위 내 미생물 균총 변화와 고창증 발병간의 상관관계 연구)

  • Kim, Sang Bum;Oh, Jong Seok;Jeong, Ha Yeon;Jung, Young Hun;Park, Beom Young;Ha, Seung Min;Im, Seok Ki;Lee, Sung Sill;Park, Ji Hoo;Park, Seong Min;Kim, Eun Tae
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.38 no.2
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    • pp.106-111
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    • 2018
  • This study was conducted to investigate the relationship between changes of rumen microflora and bloat in Jersey cow. Jersey cows (control age: 42 months, control weight: 558kg; treatment age: 29 months, treatment weight 507kg) were fed on the basis of dairy feeding management at dairy science division in National Institute of Animal Science. The change of microbial population in rumen was analyzed by using next generation sequencing (NGS) technologies due to metabolic disease. The diversity of Ruminococcus bromii, Bifidobacterium pseudolongum, Bifidobacterium merycicum and Butyrivibrio fibrisolvens known as major starch fermenting bacteria was increased more than 36-fold in bloated Jersey, while cellulolytic bacteria community such as Fibrobacter succinogenes, Ruminococcus albus and Ruminococcus flavefaciens was increased more than 12-fold in non-bloated Jersey. The proportion of bacteroidetes and firmicutes was 33.4% and 39.6% in non-bloated Jersey's rumen, while bacteroidetes and firmicutes were 24.9% and 55.1% in bloated Jersey's. In conclusion, the change of rumen microbial community, in particular the increase in starch fermenting bacteria, might have an effect to occur the bloat in Jersey cow.

Isolation of a New Agar Degrading Bacterium, Maribacter sp. SH-1 and Characterization of its Agarase (신규 한천분해세균 Maribacter sp. SH-1의 분리 및 효소 특성조사)

  • Lee, Chang-Eun;Lee, Sol-Ji;Lee, Dong-Geun;Lee, Sang-Hyeon
    • Microbiology and Biotechnology Letters
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    • v.44 no.2
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    • pp.156-162
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    • 2016
  • In this study, we isolated a new agar-degrading marine bacterium and characterized its agarase. An agardegrading marine bacterium SH-1 was isolated from seawater, collected from the seashore of Namhae in Gyeongnam province, Korea, and cultured in marine agar 2216 media. It was identified as Maribacter. sp. SH-1 by phylogenetic analyses, based on 16S rRNA gene sequence. The extracellular agarase was extracted from culture media of Maribacter sp. SH-1 and characterized. Its relative activities were 56, 62, 94, 100, and 8% at 20, 30, 40, 50, and 60℃, respectively, whereas 15, 100, 60, and 21% relative activities were observed at pH 5, 6, 7, and 8, respectively. Its extracellular agarase exhibited maximum activity (231 units/l) at pH 6.0 and 50℃, in 20 mM Tris-HCl buffer. Therefore, this agarase would be applicable as it showed the maximum activity at the temperature at which the agar is in a sol state. Furthermore, the agarase activities remained over 90% at 20, 30, and 40℃ after 0.5 h exposure at these temperatures. Thin layer chromatography analysis suggested that Maribacter sp. SH-1 produces extracellular β-agarase, as it hydrolyzes agarose to produce neoagarooligosaccharides, such as neoagarohexaose (34.8%), neoagarotetraose (52.2%), and neoagarobiose (13.0%). Maribacter sp. SH-1 and its β-agarase would be useful for the production of neoagarooligosaccharides, which shows functional properties, like skin moisturizing, skin whitening, inhibition of bacterial growth, and delay in starch degradation.

Genetic Diversity and Population Structure of Codium fragile (SURINGAR) HARlOT in Korea Using Allozymes (알로자임을 이용한 청각의 유전적 다양성과 집단구조)

  • Lee Bok-Kyu;Park So-Hye;Heo Youn-Seong;Ju Mu-Teol;Choi Joo-Soo;Huh Man-Kyu
    • Journal of Life Science
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    • v.16 no.2 s.75
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    • pp.213-218
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    • 2006
  • The study of genetic diversity and population structure was carried out in the Codium fragile using allozyme analysis. Although this species has been regarded as a ecologically and economically important source, there is no report on population structure in Korea. Starch gel electrophoresis was used to investigate the allozyme variation and genetic structure of four Korean populations of this species. Of the 15 genetic loci surveyed, nine (60.0%) was polymorphic in at least one population. Genetic diversity was high at the species level ($H_{ES}$=0.144), and, that of the population level was relatively low ($H_{EP}$=0.128). Nearly 87% of the total genetic diversity in C. fragile was apportioned within populations. The predominant asexual reproduction, population fragmentation, low fecundity, geographic isolation and colonization process are proposed as possible factors contributing to low genetic diversity in this species. The indirect estimated of gene flow based on $G_{ST}$ was 1.69. The moderate level of gene flow in C. fragile populations is mainly caused by thallus developed from isolated utricles dispersal via sea current.