• Title/Summary/Keyword: Bacillus sp. I-5

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Enzymatic Production of Amylopectin Cluster Using Cyclodextrin Glucanotransferase (Cyclodextrin Glucanotransferase를 이용한 아밀로펙틴 클러스터의 생산)

  • Lee, Hye-Won;Jeon, Hye-Yeon;Choi, Hyejeong;Shim, Jae-Hoon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.9
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    • pp.1388-1393
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    • 2014
  • To enzymatically prepare amylopectin cluster (APC), cyclodextrin glucanotransferase (CGTase I-5) and its mutant enzyme from alkalophilic Bacillus sp. I-5 were employed, after which the hydrolysis patterns of CGTase wild-type and its mutant enzyme toward amylopectin were investigated using multi-angle laser light scattering. CGTase wild-type dramatically reduced the molecular weight of waxy rice starch at the initial reaction, whereas the mutant enzyme degraded waxy rice starch relatively slowly. Based on the results, the molecular weight of one cluster of amylopectin could be about $10^4{\sim}10^5g/mol$. To determine production of cyclic glucans from amylopectin, matrix-assisted laser desorption ionization time-of-flight mass spectrometry was performed. CGTase I-5 produced various types of cyclic maltooligosaccharides from amylopectin, whereas the mutant enzyme hardly produced any.

Role of Crops and Residues, and Fertilization to Changes of microbial Population, Soil chemical Properties and Plant Growth -I. Microbial Population in the Habitate (작물(作物)의 종류(種類)와 잔사(殘渣) 및 시비량(施肥量)이 토양미생물상(土壤微生物相), 이화학성(理化學性) 및 작물생육(作物生育)에 미치는 역할(役割) -I. 미생물상(微生物相) 변화(變化))

  • Kim, Seung-Hwan;Lee, Sang-Kyu
    • Korean Journal of Soil Science and Fertilizer
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    • v.25 no.4
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    • pp.370-377
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    • 1992
  • A series of field and laboratory experimentes were conducted to find out the effects of kinds of crops residues and the different amount of fertilizer to the microbial interaction, chemical properties, plant growth and their interaction under continous cultivation of hot pepper, soybean, sesame and upland rice. The results obtained were summerized follows : 1. Total number of bacteria and actinomycetes were enhanced by cultivation of upland rice and soybean while no appreciable effects were obtained by the cultivation of hot pepper and sesame. 2. The number of bacteria, actinomycetes and fungi were increased by return of crop residues when the cultivation of hot pepper, soybean, sesame and upland rice. Specially, actinomycetes was remarkably increased by upland rice cultivation. 3. Increased amount of fertilizer were remarkably affected to decrease of number of soil microorganisms. Specially, actinomycetes succession was appearently affects while plant growing time. 4. The number of identified soil bacterial species were obtained high in order of Bacillus sp.>Rhizobium sp.>Agrobacterium sp.>Pseudomonas sp. The number of Gram positive bacteria were superior than that of Gram negative bateria. 5. Interaction between microbial succession and crops cultivation, the number of Bacillus sp. was increased in hot pepper, Rhizobium sp. was in soybean, and sesame, and Agrobacterium sp. were increased in soybean, respectively. 6. Survival and habitate of soil microorganisms were differ with kinds of crop, application of residue and fertilizers. Most high number of Bacillus sp. Rhizobium sp. and Pseudomonas sp. were obtained on the rhizoplane and rhizosphere while Agrobacterium sp. and Fusarium sp. were high in rhizosphere. 7. Factors in relation to change of soil microbial population was obtained high in order of climates>crops>organic>matter>fertilizer.

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Characterization of an Extracellular Xylanase from Bacillus sp. HY-20, a Bacterium in the Gut of Apis mellifera (꿀벌(Apis mellifera)의 장내 세균인 Bacillus sp. HY-20이 분비하는 Xylanase의 특성)

  • Lee, Lan-Hee;Kim, Do-Young;Han, Mi-Kyoung;Oh, Hyun-Woo;Ham, Su-Jin;Park, Doo-Sang;Bae, Kyung-Sook;Sok, Dai-Eun;Shin, Dong-Ha;Son, Kwang-Hee;Park, Ho-Yong
    • Korean Journal of Microbiology
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    • v.45 no.4
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    • pp.332-338
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    • 2009
  • A xylan-decomposing bacterium, HY-20, was isolated from the gut of a honeybee, Apis mellifera, and identified as Bacillus sp. The extracellular GH11 xylanase (XylP) gene (687-bp) of strain HY-20 encoded a protein of 228 amino acids with a deduced molecular mass of 25,522 Da and a calculated pI of 9.33. The primary structure of XylP was 97% identical to that of B. pumilus xylanase (GenBank accession no.: AY526092) that has not been characterized yet. The recombinant His-tagged enzyme (rXylP) overexpressed in Escherichia coli BL21 harboring pET-28a(+)/xylP was purified to electrophoretic homogeneity by cation exchange and gel permeation chromatographies. The purified enzyme exhibited the highest catalytic activity toward birchwood xylan at pH 6.5 and $50^{\circ}C$ and retained approximately 50% of its original activity when pre-incubated at $55^{\circ}C$ for 15 min. The recombinant enzyme was completely inactivated by $Hg^{2+}$ (1 mM) and N-bromosuccinimide (5 mM), while its activity was slightly stimulated by approximately 10% in the presence of $Mn^{2+}$ (1 mM), $Fe^{2+}$ (1 mM), and sodium azide (5 mM). rXylP was able to efficiently degrade various polymeric xylose-based substrates but PNP-sugar derivatives and glucose-based polymers were not susceptible to the enzyme.

Treatment of an Authentic Textile-dyeing Wastewater Utilizing a Fluidized Biofilter and Hybrid Recirculating System Composed of the Fluidized Biofilter and a UV/photocatalytic Reactor (실제 혼합염색폐수의 유동상 시스템을 활용한 미생물처리와 하이브리드 재순환시스템처리)

  • Lee, Eun Ju;Lim, Kwang-Hee
    • Korean Chemical Engineering Research
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    • v.53 no.1
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    • pp.71-77
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    • 2015
  • A fluidized biofilter was filled with Pseudomonas sp. and Bacillus cereus/thuringiensis-fixed waste-tire crumb media and was run to treat authentic textile-dyeing wastewater mixed with alkaline polyester-weight-reducing wastewater. As a result, its removal efficiency of $COD_{Cr}$ and color were 75~80% and 67%, respectively. In addition, upon constructing hybrid-recirculating system composed of the fluidized biofilter and a 450 W-UV/photocatalytic reactor, only fluidized biofilter was run bypassing UV/photocatalytic reactor at stage I. Subsequently, the hybrid system was continuously run at stage II-i, ii and iii. At stage II-i, the total removal efficiency of $COD_{Cr}$ was enhanced to be 80~85%, compared to 75% at stage I, owing to 20~30% removal efficiency of the UV/photocatalytic reactor. However, at stage II-i, the total removal efficiency of color was enhanced to be 65~70%, compared to 45~65% at stage I, even though the removal efficiency of the UV/photocatalytic reactor was tantamount to merely 0~5%. As far as the removal efficiency of fluidized biofilter of the hybrid-recirculating system is concerned, its removal efficiency of color was enhanced by the synergy effect of the hybrid-recirculating system unlike $COD_{Cr}$. Besides, despite of the increase of hybrid-recirculating system-recycle ratio, the deactivation of photo-catalytic activity was scarcely observed to eliminate the color while its irreversible deactivation was observed to eliminate $COD_{Cr}$.

Analysis of Cellular Fatty Acid Methyl Esters (FAMEs) for the Identification of Bacillus anthracis (균체 지방산 분석을 이용한 Bacillus anthracis의 동정)

  • Kim, Won-Yong;Song, Tae-Wook;Song, Mi-Ok;Nam, Ji-Yeon;Park, Chul-Min;Kim, Ki-Jung;Chung, Sang-In;Choi, Chul-Soon
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.1
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    • pp.31-40
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    • 2000
  • Bacillus anthracis, the etiological agent of anthrax has been classified into the Bacillus subgroup I with B. cereus, B. mycoides and B. thuringiensis based on morphological and DNA similarity. DNA studies have further indicated that these species have very AT-rich genomes and high homology, indeed it has been proposed that these four sub-species be recognized as members of the one species. Several methods have been developed to obtain good differentiation between these species. However, none of these methods provides the means for an absolutely correct differntiation. The analysis of fatty acid methyl esters (FAMEs) was employed as a quick, simple and reliable method for the identification of 21 B. anthracis strains and closley related strains. The most significant differences were found between B. anthracis and B. anthracis closely related strains in FAMEs profiles. All tested strains of B. anthracis had a branched fatty acid C17:1 Anteiso A, whereas the fraction of unsaturated fatty acid Iso C17:1 w10c was found in B. anthracis closely related strains. By UPGMA clustering analysis of FAMEs profiles, all of the tested strains were classified into two clusters defined at Euclidian distance value of 24.5. The tested strains of B. anthracis were clustered together including Bacillus sp. Kyungjoo 3. However, the isolates of B. anthracis closely related spp. Rho, S10A, 11R1, CAU9910, CAU9911, CAU9912 and CAU9913 were clustered with the other group. On the basis of these results, isolates of B. anthracis Bongchon, Kyungjoo 1, 2 and Bacillus sp. Kyungjoo 3 were reclassified as a B. anthracis. It is concluded that FAMEs analysis provides a sensitive and reliable method for the identification of B. anthracis from closely related taxa.

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Characterization of Denitrifying and Dissimilatory Nitrate Reduction to Ammonium Bacteria Isolated from Mud Crab Culture Environment

  • Hastuti, Yuni Puji;Rusmana, Iman;Nirmala, Kukuh;Affandi, Ridwan;Fatma, Yuli Siti
    • Microbiology and Biotechnology Letters
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    • v.49 no.3
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    • pp.432-439
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    • 2021
  • Microbial community plays important roles in the culture environment of mud crab Scylla serrata. One of the environmental management efforts for the cultivation of S.serrata is by stabilizing microorganisms involved in nitrogen cycle process. The availability of dissolved inorganic nitrogen in its culture environment under a recirculating system closely relates to the nitrogen cycle, which involves both anaerobic and aerobic bacterial activities. Anaerobically, there are two major nitrogen compound degradation processes, i.e., denitrification and dissimilatory nitrate reduction to ammonium (DNRA). This study aimed to identify denitrifying and DNRA bacteria isolated from the recirculating cultivation of S. serrata. The water samples were collected from anaerobic filters called close filter system, which is anaerobically conditioned with the addition of varying physical filter materials in the recirculating mud crab cultures. The results showed that three denitrifying bacterial isolates and seven DNRA bacterial isolates were successfully identified. The phylogenetic analysis based on 16S rRNA gene of the denitrifying bacteria revealed that HIB_7a had the closest similarity to Stenotrophomonas daejeonensis strain MJ03. Meanwhile, DNRA bacterial isolate of HIB_92 showed a 100% similarity to Bacillus sonorensis strain N3, Bacillus vallismortis strain VITS-17, Bacillus tequlensis strain TY5, Geobacillus sp. strain DB24, Bacillus subtilis strain A1, and Bacillus mojavensis strain SSRAI21. This study provides basic information denitrifying and DNRA bacterial isolates identity which might have the potential to be applied as probiotics in aquaculture systems in order to maintain optimal environmental conditions.

Improvement in Antagonistic Ablility of Antagonistic Bacterium Bacillus sp. SH14 by Transfer of the Urease Gene. (Urease gene의 전이에 의한 길항세균 Bacillus sp. SH14의 길항능력 증가)

  • 최종규;김상달
    • Microbiology and Biotechnology Letters
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    • v.26 no.2
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    • pp.122-129
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    • 1998
  • It were reported that antifungal mechanism of Enterobacter cloacae is a volatile ammonia that produced by the strain in soil, and the production of ammonia is related to the bacterial urease activity. A powerful bacterium SH14 against soil-borne pathogen Fusarium solani, which cause root rot of many important crops, was selected from a ginseng pathogen suppressive soil. The strain SH14 was identified as Bacillus subtilis by cultural, biochemical, morphological method, and $API^{circledR}$ test. From several in vitro tests, the antifungal substance that is produced from B. subtilis SH14 was revealed as heat-stable and low-molecular weight antibiotic substance. In order to construct the multifunctional biocontrol agent, the urease gene of Bacillus pasteurii which can produce pathogenes-suppressive ammonia transferred into antifungal bacterium. First, a partial BamH I digestion fragment of plasmid pBU11 containing the alkalophilic B. pasteurii l1859 urease gene was inserted into the BamH I site of pEB203 and expressed in Escherichia coli JM109. The recombinant plasmid was designated as pGU366. The plasmid pGU366 containing urease gene was introduced into the B. subtilis SH14 with PEG-induced protoplast transformation (PIP) method. The urease gene was very stably expressed in the transformant of B. subtilis SH14. Also, the optimal conditions for transformation were established and the highest transformation frequency was obtained by treatment of lysozyme for 90 min, and then addition of 1.5 ${mu}g$/ml DNA and 40% PEG4000. From the in vitro antifungal test against F. solani, antifungal activity of B. subtilis SH14(pGu366) containing urease gene was much higher than that of the host strain. Genetical development of B. subtilis SH14 by transfer of urease gene can be responsible for enhanced biocontrol efficacy with its antibiotic action.

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Overexpression of Termostable Bacillus sp. in Recombinant E.coli (재조합 E.coli에서 고온성 Bacillus 균주의 과발현에 관한 연구)

  • 서화정;이인선
    • Journal of Food Hygiene and Safety
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    • v.15 no.1
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    • pp.51-54
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    • 2000
  • In the 5'-flanking region of the D-AAT, AspAT and AlaDH gene, I found three or two pairs of sequences(designated as Pl, P2, P3) which show significant similarity to the E.coli consensus sequences of -35 and -10 for promoters. The spacing between -35 and -10 is 16 to 18bp in all the three putative promoters Pl, P2 and P3 which is in good agreement with the preferred spacer length in E.coli and in B.subtilis. Therefore, the putative promoters may also function to increase the efficiency of transcriptional initiation. The most stable, double-helical“Shine-Dalgarno”pairing is formed with a free energy change(ΔG) of -13.0 kcal/mol, -9.6 kcal/mol, -15.8 kcal/mol.

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Isolation and Identification of a Streptomyces sp. that Produces Antibiotics Against Multidrug - Resistant Acinetobacter baumannii (다제내성 Acinetobacter baumannii의 생장을 억제하는 항생물질을 생산하는 방선균의 분리.동정 및 항균효과)

  • Rhee, Ki-Hyeong
    • Microbiology and Biotechnology Letters
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    • v.39 no.1
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    • pp.37-42
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    • 2011
  • I isolated the actinomycete strain KH223 from soil samples collected from the Kye Ryong mountain area. This strain is antagonistic to the multidrug-resistant Acinetobacter baumannii. KH223 was confirmed as belonging to the genus Streptomyces based on the scanning electronmicroscopy(SEM) observations of the diaminopimelicacid(DAP) type and morphological and physiological characteristics. Comparison of the 16S rDNA nucleotide sequences revealed that KH223 has a relationship with Streptomyces galbus. Production of antibiotics by KH223 was most favorable when cultured on a glucose, polypeptone, and yeast extract(PY) medium for 6 days at 27$^{\circ}C$. The supernatant was found to exhibit an antimicrobial effect on various kinds of bacteria and fungi. Particularly, butanol and ethylacetate extracts of KH223 and cyclo(trp-trp) exhibited significant activity against A. baumannii at concentration ranges of 0.8-12.5 ${\mu}g$/mL, 5.0-25 ${\mu}g$/mL and 12.5${\rightarrow}$100 ${\mu}g$/mL, respectively. Moreover, in contrast to cyclo(trp-trp) had shown to activity against Micrococcus luteus JCM 1464 at the concentration of 12.5 ${\mu}g$/mL, the butanol extract of KH223 showed significant activity against Bacillus subtilis IAM 1069 and Micrococcus luteus JCM 1464 at the concentration of 0.4 and 0.8 ${\mu}g$/mL, respectively. These results suggest that KH223 may have a great potential in the production of new antibiotics to combat multidrug-resistant pathogens and further studies may be warranted for the same.

Reevaluation of Isolation and Identification of Gram-positive Bacteria in Kimchi (김치에 서식하는 Gram 양성세균의 분리 및 동정의 재평가)

  • 임종락;박현근;한홍의
    • Korean Journal of Microbiology
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    • v.27 no.4
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    • pp.404-414
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    • 1989
  • Attempts were made to isolate and identify Gram-positive or lactic acid bacteria in Kimchi fermentation. Species diversity depended on isolation media and temperatures, and diversity tended to be reduced with decrease of temperature. MRS and KM (natural medium prepared from Kimchi materials) were suitable respectively for isolation and present number of species. Identification of isolates was performed by dichotomous identification schemes arranged on the basis of Bergey's manual of Systematic Bacteriology (1986). Gram-positive bacteria isolated at different temperatures (5, 15, $25^{\circ}C$) were 5 species of Leuconostoc, 4 species of Streptococcus, 3 species of Pediococcus, 2 species of Bacillus and 18 species of Lactobacillus. Species with high frequency of appearance were Lactobacillus plantarum, Streptococcus raffinolactis, Leuconostoc mesenteroides subsp. mesenteroides at $25^{\circ}C$, L. plantarum, Lactobacillus fructosus, L. mesenteroides subsp. mesenteroides at $15^{\circ}C$ and L. mesenteroides subsp. mesenteroides, Leuconosotoc paramesenteroides, Lactobacillus maltaromicus at $15^{\circ}C$. In general, Kimchi fermentation was achieved by Lactobacillus spp. (59.7% frequency) at $25^{\circ}C$ and Leuconostoc spp. (65.2% frequency) at $5^{\circ}C$. Pediococcus cerevisiae and Streptococcus faecalis which have been so far known as bacteria of Kimchi fermentation were not isolated.

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