• Title/Summary/Keyword: Bacillus sp. N16-5

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Isolation and Characteristics of Biosurfactant Producing Bacterium, Bacillus sp. TBM 911-5 (Biosurfactant 생산균주 Bacillus sp. TBM 911-5의 분리 및 특성)

  • 김선희;정연주;이상철;유주순;주우홍;정수열;최시림;최용락
    • Journal of Life Science
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    • v.14 no.2
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    • pp.320-324
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    • 2004
  • The objective of this study was investigate the characteristic of biosurfactant produced from the isolated strain. The strain was isolated from soil samples and identified as Bacillus sp. TBM 911-5 by physiological characteristics and the partial nucleotide sequence analysis of 16S rDNA. We measured the surface tension every 6 hours for 80 hours. The surface tension of the culture filtrate of Bacillus sp. TBM 911-5 was decreased to 29 mN/m. Biosurfactant concentration was determined by diluting the culture filtrate until the critical micelle concentration (CMC). The biosurfactant emulsified hydrocarbons, vegetable oil and crude oil. Using soybean oil as substrate, the maximum emulsification activity and stability was obtained from the biosurfactant. The biosurfactant produced from Bacillus sp. TBM 911-5 had strong properties as an emulsifying agent and an emulsion-stabilizing agent.

Intrageneric Protoplast Fusion between Alkalophilic Bacillus sp. F204 and Bacillus sp. K 17 (호알칼리성 Bacillus sp. F204와 Bacillus sp. K 17의 원형질체 융합)

  • 성낙계;노종수;박석규;정영철
    • Microbiology and Biotechnology Letters
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    • v.16 no.4
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    • pp.275-281
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    • 1988
  • To develop cellulase and xylanase-producing strain by protoplast fusion, alkalophilic Bacillus sp. F204 and K17 were treated with NTG(N-methyl-N'-nitro-N-nitrosoguanidine) and isolated anti-biotics resistant strains of S20 (Km$^r$ , Cm$^r$) and G70 (Str$^r$). The frequency of protoplast formation was about 95% when cells of mid-log phase were treated with 200$\mu\textrm{g}$/ml Iysozyme at 37$^{\circ}C$ for 30-45 minutes. Under addition of 0.4-0.5M sodium succinate, 0.5% casamino acid, 1.5% polyvinylpyrrolidone, 25mM MgC1$_2$ and 50mM CaC1$_2$ to the regeneration medium, the regeneration frequency of Bacillus sp. F204 and K17 was 24.9% and 26.2%, respectively. The fusion frequency was 6.6$\times$10$^{-6}$ in the presence of 30% polyethylene glycol 6000 containing 50mM $Ca^{++}$ at 45$^{\circ}C$ for 5 minutes. Cellulase complex and xylanase activities of fusant were compared with parental strains.

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Cloning, Expression, and Characterization of a Highly Active Alkaline Pectate Lyase from Alkaliphilic Bacillus sp. N16-5

  • Li, Gang;Rao, Lang;Xue, Yanfen;Zhou, Cheng;Zhang, Yun;Ma, Yanhe
    • Journal of Microbiology and Biotechnology
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    • v.20 no.4
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    • pp.670-677
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    • 2010
  • An alkaline pectate lyase, Bsp165PelA, was purified to homogeneity from the culture broth of alkaliphilic Bacillus sp. N16-5. The enzyme showed a specific activity as high as 1,000 U/mg and had optimum activity at pH 11.5 and $50^{\circ}C$. It was composed of a single polypeptide chain with a molecular mass of 42 kDa deduced from SDS-PAGE, and its isoelectric point was around pH 6.0. It could efficiently depolymerize polygalacturonate and pectin. Characterization of product formation revealed unsaturated digalacturonate and trigalacturonate as the main products. The pectate lyase gene (pelA) contained an open reading frame (ORF) of 1,089 bp, encoding a 36-amino acids signal peptide and a mature protein of 326 amino acids with a calculated molecular mass of 35.943 Da. The deduced amino acid sequence from the pelA ORF exhibited significant homology to those of known pectate lyases in polysaccharide lyase family 1. Some conserved active-site amino acids were found in the deduced amino acid sequence of Bsp165PelA. $Ca^{2+}$ was not required for activity on pectic substrates.

Bacillus ginsengihumi sp. nov., a Novel Species Isolated from Soil of a Ginseng Field in Pocheon Province, South Korea

  • Ten Leonid N.;Im Wan-Taek;Baek Sang-Hoon;Lee, Jung-Sook;Oh, Hee-Mock;Lee, Sung-Taik
    • Journal of Microbiology and Biotechnology
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    • v.16 no.10
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    • pp.1554-1560
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    • 2006
  • A Gram-positive, aerobic or facultative anaerobic, non motile, endospore-forming bacterial strain, designated Gsoil $114^T$, was isolated from a soil sample of a ginseng field in Pocheon Province (South Korea), and was characterized taxonomically by using a polyphasic approach. It grew well on nutrient agar medium and utilized a limited number of organic substrates as sole carbon sources, including D-xylose and some other carbohydrates, but did not utilize L-amino acids and organic acids. The isolate was positive for oxidase test but negative for catalase, and negative for degradation of macromolecules such as starch, cellulose, xylan, casein, chitin, and DNA. The G+C content of the genomic DNA was 41.8 mol%. The predominant isoprenoid quinone was menaquinone 7 (MK-7). The major fatty acids were $anteiso-C_{15:0}$ (32.1%), $iso-C_{15:0}$ (30.5%), and $anteiso-C_{17:0}$ (30.2%). Comparative 16S rRNA gene sequence analysis showed that strain Gsoil $114^T$ fell within the radiation of the cluster comprising Bacillus species and joined Bacillus shackletonii LMG $18435^T$ with a bootstrap value of 95%. The highest 16S rRNA gene sequence similarities were found with Bacillus shackletonii LMG $18435^T$ (97.6%), Bacillus acidicola DSM $14745^T$ (96.9%), Bacillus sporothermodurans DSM $10599^T$ (96.5%), and Bacillus oleronius DSM $9356^T$ (96.5%). The phylogenetic distance from any other validly described species within the genus Bacillus was less than 96%. DNA-DNA hybridization experiments showed that the DNA-similarities between strain Gsoil $114^T$ and closest phylogenetic neighbors were less than 39%. On the basis of its phenotypic properties and phylogenetic distinctiveness, strain Gsoil $114^T$ (=KCTC $13944^T$=DSMZ $18134^T$) was classified in the genus Bacillus as the type strain of a novel species, for which the name Bacillus ginsengihumi sp. nov. is proposed.

Improvement effect of total nitrogen and amino acid content in spent mushroom substrates by bacterial treatment (세균을 이용한 수확후배지의 총질소 및 아미노산 증진 효과)

  • Baek, Il-Sun;Kim, Jeong-Han;Lee, Yong-Seon;Shin, Bok-Eum;Lee, Young-Soon
    • Journal of Mushroom
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    • v.16 no.3
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    • pp.225-230
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    • 2018
  • This study was conducted to reuse spent mushroom substrates (SMS) of Pleurotus ostreatus and improve their nitrogen content by bacterial treatment. Two kinds of bacteria were used to investigate the increase in total nitrogen (T-N) content. Bacillus sp. (GM20-4) was isolated from SMS of oyster mushroom, and Rhodobacter sphaeroides (RS) was obtained from Gwangju Si Agricultural Technology Center. SMS samples were collected from three oyster mushroom cultivation farms located in Gyeonggi-do province, Korea. When dried SMS was inoculated with 30% culture broth of GM20-4 and RS and incubated at room temperature ($25{\pm}2^{\circ}C$) for 5 days, T-N content increased. To investigate the T-N content of other SMS, three dried SMS samples (A, B, and C) were treated by the same method using GM20-4 and RS. As a result, the T-N content of sample B was 20% higher than that of the control, whereas the T-N content of samples A and C increased to 17% and 12%, respectively. The change in T-N content by bacterial treatment of wet SMS was slightly higher than that of the control. The changes in amino acid content were also found to be higher than those in the control in all SMS samples by GM20-4 and RS treatment. Aspartic acid and glutamic acid contents were the highest among all amino acid compositions. Especially, the aspartic and glutamic acid contents of sample B increased by 2.9 folds higher than the control.

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.

Changes in Physicochemical Properties and Microbial Population during Fermenting Process of Organic Fertilizer (혼합발효 유기질비료의 발효과정 중 이화학성 및 미생물밀도 변화)

  • Lee, Jong-Tae;Lee, Chan-Jung;Kim, Hee-Dae
    • Korean Journal of Soil Science and Fertilizer
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    • v.37 no.2
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    • pp.116-123
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    • 2004
  • This study was conducted to investigate the changes in physicochemical and microbiological properties during fermenting process of organic fertilizer which was made from the mixture of organic materials such as sesame oil cake, fish meal, blood meal, rice bran, ground bone meal, and natural minerals such as illite, crusted oyster shell and loess. They were mixed and fermented for 70 days. The sesame oil cake and rice bran, major ingredients for organic fertilizers, consisted of 7.6 and 2.6% total nitrogen, 3.6 and 4.6% $P_2O_5$, 1.4 and 2.2% $K_2O$, respectively. The ground bone meal included 29.2% $P_2O_5$ and illite included 3.8% $K_2O$. Temperature of organic fertilizer during the fermentation rapidly increased over $50^{\circ}C$ within 2 days after mixing and stabilized similar to outdoor temperature after 40 days. Moisture content decreased from 36.3 to 16.0% after 1 month. C/N ratio of organic fertilizer slightly increased until 30 days and thereafter, it slowly decreased, It resulted from the faster decrease of total nitrogen concentration compared with organic matter. Concentration of $NH_4-N$ in organic fertilizer rapidly increased from 1,504 to $5,530mg\;kg^{-1}$, the highest concentration after 10 days. Meantime, $NO_3-N$ concentration was low and constant about $150mg\;kg^{-1}$ over the whole fermenting period. This result seemed to be due to the high pH. The organic ferfilizer fermented for 70 days was composed of 2.7% N, 2.8% $P_2O_5$, 1.8% $K_2O$, and 35.9% organic matter. Total populations of aerobic bacteria, Bacillus sp. and actinomycetes, after fermenting process, were $12.5{\times}10^{10}$, $45.5{\times}10^{5}$ and $13.6{\times}10^{5}cfu\;g^{-1}$ respectively. Pseudomonas sp. was $71.9{\times}10^{7}cfu\;g^{-1}$ at first, but it rapidly decreased according to the rise of temperature. Yeasts played an important role in the early stage of fermentation and molds did in the late stage.

Changes of Microbial Community Depending on Different Dissolved Oxygen in Biological Nitrogen Removal Process (생물학적 질소제거 공정에서 용존산소변화에 따른 미생물의 군집변화)

  • Park, Jong-Il;Lee, Tae-Jin
    • Journal of Korean Society of Environmental Engineers
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    • v.30 no.9
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    • pp.939-947
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    • 2008
  • PCR-DGGE method was applied to analyze changes of microbial community in simultaneous nitrification and denitrification (SND) bioreactor with various DO concentrations. In the analysis of eubacterial community, band profiles of DGGE were similar with 2 or 1 mg/L DO concentrations in the reactor. Experimental results led to 16 different bacteria being identified, including 5 dominant strains(3 strains of Uncultured Bacterium, 1 strains of Bacillus, 1 strains of Uncultured Bacteroidetes). DGGE results at 0.5 mg/L DO concentration led to 12 strains being identified, including 7 dominant strains(5 strains of Uncultured Bacterium, 2 strains of Zoogloea sp.). DGGE results at 0.1 mg/L DO concentration led to 11 strains being identified, including 3 dominant strains(1 strains of Uncultured Bacterium, 2 strains of Zoogloea sp.). In DGGE band profiles of $\beta$-AOB($\beta$-Ammonia Oxidizing Bacteria), only one band was observed. This band had 97% similarity with Nitrosomonas sp. done DNB Y20. This band was clearly observed at the 2, 1 and 0.5 mg/L DO concentrations, while the brightness of the band at 0.1 mg/L DO concentration was mostly dimmed. In DGGE band profiles of denitrification process, 5 bands(3 strains of Uncultured organism containing nirS, 2 strains of Uncultured organism containing nirK) were observed. Among those bands, the brightness of one band was gradually increased at the lower DO concentrations. This band has 86% identity with Uncultured organism clone eS1 cd1 nirS gene, partial cds. Based on this result, it could be concluded that Uncultured organism clone eS1 cd1 nirS gene, partial cds is a predominant microorganism in the denitrification process.

Diffusible and Volatile Antifungal Compounds Produced by an Antagonistic Bacillus velezensis G341 against Various Phytopathogenic Fungi

  • Lim, Seong Mi;Yoon, Mi-Young;Choi, Gyung Ja;Choi, Yong Ho;Jang, Kyoung Soo;Shin, Teak Soo;Park, Hae Woong;Yu, Nan Hee;Kim, Young Ho;Kim, Jin-Cheol
    • The Plant Pathology Journal
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    • v.33 no.5
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    • pp.488-498
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    • 2017
  • The aim of this study was to identify volatile and agardiffusible antifungal metabolites produced by Bacillus sp. G341 with strong antifungal activity against various phytopathogenic fungi. Strain G341 isolated from four-year-old roots of Korean ginseng with rot symptoms was identified as Bacillus velezensis based on 16S rDNA and gyrA sequences. Strain G341 inhibited mycelial growth of all phytopathogenic fungi tested. In vivo experiment results revealed that n-butanol extract of fermentation broth effectively controlled the development of rice sheath blight, tomato gray mold, tomato late blight, wheat leaf rust, barley powdery mildew, and red pepper anthracnose. Two antifungal compounds were isolated from strain G341 and identified as bacillomycin L and fengycin A by MS/MS analysis. Moreover, volatile compounds emitted from strain G341 were found to be able to inhibit mycelial growth of various phytopathogenic fungi. Based on volatile compound profiles of strain G341 obtained through headspace collection and analysis on GC-MS, dimethylsulfoxide, 1-butanol, and 3-hydroxy-2-butanone (acetoin) were identified. Taken together, these results suggest that B. valezensis G341 can be used as a biocontrol agent for various plant diseases caused by phytopathogenic fungi.

Studies on the Fermentation on Soy Sauce by Bacteria (세균(細菌)을 이용(利用)한 간장 제조(製造)에 관(關)한 연구(硏究))

  • Ju, Hyun-Kyu;Ro, Sin-Kyu;Im, Moo-Hyun
    • Korean Journal of Food Science and Technology
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    • v.4 no.4
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    • pp.276-284
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    • 1972
  • A strain of Bacillus sp. which has powerful enzymatic activity and sets free a little amount of ammonia nitrogen and large amount of amino nitrogen was separated from Meju, Denzang, Chungkukjang, and paddy straw to make the soy-sauce rapidly and conveniently in the various mixing ratio of steamed soy-bean and wheat parched or steamed. Total nitrogen and amino nitrogen were increased during the maturing. The acidity of soy-sauce was increased to $1.15{\sim}1.98%$ than ordinary soy-sauce. pH were ranged in $4.6{\sim}6.0%$. The fermenting time takes 30 days while it takes at least 4 monthes in ordinary method. The best results were obtained with the mixed ratio of 1 : 1 or 1 : 2 (wheat : soy-bean).

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