• Title/Summary/Keyword: Bacillus subtilis spore

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The Possible Role of SCO3388, a tmrB-like Gene of Streptomyces coelicolor, in Germination and Stress Survival of Spores

  • Kwon, So-Yeon;Kwon, Hyung-Jin
    • Journal of Applied Biological Chemistry
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    • v.56 no.3
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    • pp.165-170
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    • 2013
  • The SCO3388 gene from Streptomyces coelicolor is homologous to tmrB, the tunicamycin resistance gene of Bacillus subtilis. The SCO3388-inactivation strain (SY-tbl-1) was generated by replacing SCO3388 with thiostrepton resistance gene. Spores of S. coelicolor derivatives were prepared on mannitol-soy flour (MS) agar on which SY-tbl-1 displayed no significant defect in growth and development. When plated on R4 agar, spores of SYtbl-1 displayed retardation in growth and sporulation, whereas its mycelium gave rise to normal growth. Thus, SCO3388 is suggested to be involved in the dormant spore germination. Expression of SCO3388 under the ermE1 promoter restored but only partially the ability to sporulate in SY-tbl-1. Neither SY-tbl-1 nor SY-tbl-1/ermE1p-SCO3388 showed a difference in tunicamycin resistance to the wild type whereas, interestingly, the introduction of ermE1p-SCO3388 dramatically enhanced spore survival to heat and detergent treatments, suggesting that SCO3388 might play a role in the maintenance of spore cell wall integrity.

Isolation and characterization of a Bacillus spp. for manufacturing the feed additives in livestock (가축의 보조사료 개발을 위한 Bacillus spp.의 분리 및 특성)

  • Park, Hae Suk;Jo, Seung Wha;Yim, Eun Jung;Kim, Yun Sun;Moon, Sung Hyun;Cho, Ho Seong;Kim, Hyun-Young;Cho, Yong Sik;Cho, Sung Ho
    • Korean Journal of Microbiology
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    • v.51 no.4
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    • pp.419-426
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    • 2015
  • The aims of this study were to isolate spore-forming Bacillus strains that exhibit high digestibility and anti-pathogenic bacteria toward feed for calves. Total 136 spore-forming strains were isolated from finished feeds and their ingredients. Among them, 93 strains were identified as Bacillus species when analyzed by 16S rRNA sequencing. For industrial use, three strains named as Bacillus licheniformis SHS14, B. subtilis LCB7, B. amyloliquefaciens LCB10 were selected after evaluating the industrial standards that are related with heat and acid resistance, enzyme activities, and anti-pathogenic activities against Samonella dublin ATCC15480 and E. coli K99. After each culture, 3 selected strains were mixed together at 1:1:1 (v/v/v) ratio and then prepared as the mixed starter culture for feeding. The changes in microbial community were analyzed via 16S rRNA metagenomics. The initial community ratio among three strains was maintained even after manufacturing into final products. Also, in vitro, enzymatic and anti-pathogenic activities were almost same as those when cultured in single culture, and results of anti-pathogenic activities conducted with calves showed 90% activities against lincomycin, which would be indicative of a promising feed starter.

Isolation and Identification of Bacillus sp. LAM 97-44 Producing Antifungal Antibiotics (항진균성 항생물질을 생산하는 Bacillus sp. LAM 97-44의 분리 및 동정)

  • Lee, No-Woon;Kim, Cheon-Suk;Do, Jae-Ho;Jung, In-Chan;Lee, Hyean-Woo;Yi, Dong-Heui
    • Applied Biological Chemistry
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    • v.41 no.3
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    • pp.208-212
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    • 1998
  • In order to develop an effective antifungal antibiotics, over 700 isolates of bacteria, mold and actinomytes were screened from soil, and LAM 97-44 were selected as a strain producing the strong antifungal antibiotics against Candida albicans. Morphological, cultural and physiological characteristics of LAM 97-44 were investigated for the indentification. The cell size of LAM 97-44 was $2{\sim}3{\times}1{\sim}1.5\;{\mu}m$, and the shape of spore was of ellipsoidal. As a carbon source, LAM 97-44 utilized fructose, glucose, glycerol, maltose and raffinose but did not utilize arabinose, cellulose and xylose. The fatty acids of the cells included various iso-type and anteiso-type. Conclusively, the strain LAM 97-44 was proved to be Bacillus subtilis.

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Three Non-Aspartate Amino Acid Mutations in the ComA Response Regulator Receiver Motif Severely Decrease Surfactin Production, Competence Development, and Spore Formation in Bacillus subtilis

  • Wang, Xiaoyu;Luo, Chuping;Liu, Youzhou;Nie, Yafeng;Liu, Yongfeng;Zhang, Rongsheng;Chen, Zhiyi
    • Journal of Microbiology and Biotechnology
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    • v.20 no.2
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    • pp.301-310
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    • 2010
  • Bacillus subtilis strains produce a broad spectrum of bioactive peptides. The lipopeptide surfactin belongs to one well-known class, which includes amphiphilic membrane-active biosurfactants and peptide antibiotics. Both the srfA promoter and the ComP-ComA signal transduction system are an important part of the factor that results in the production of surfactin. Bs-M49, obtained by means of low-energy ion implantation in wild-type Bs-916, produced significantly lower levels of surfactin, and had no obvious effects against R. solani. Occasionally, we found strain Bs-M49 decreased spore formation and the development of competence. Blast comparison of the sequences from Bs-916 and M49 indicate that there is no difference in the srfA operon promoter PsrfA, but there are differences in the coding sequence of the comA gene. These differences result in three missense mutations within the M49 ComA protein. RT-PCR analyses results showed that the expression levels of selected genes involved in competence and sporulation in both the wild-type Bs-916 and mutant M49 strains were significantly different. When we integrated the comA ORF into the chromosome of M49 at the amyE locus, M49 restored hemolytic activity and antifungal activity. Then, HPLC analyses results also showed the comA-complemented strain had a similar ability to produce surf actin with wild-type strain Bs-916. These data suggested that the mutation of three key amino acids in ComA greatly affected the biological activity of Bacillus subtilis. ComA protein 3D structure prediction and motif search prediction indicated that ComA has two obvious motifs common to response regulator proteins, which are the N-terminal response regulator receiver motif and the C-terminal helix-turn-helix motif. The three residues in the ComA N-terminal portion may be involved in phosphorylation activation mechanism. These structural prediction results implicate that three mutated residues in the ComA protein may play an important role in the formation of a salt-bridge to the phosphoryl group keeping active conformation to subsequent regulation of the expression of downstream genes.

A Constitutive Inulase from a Bacillus (Bacillus 균이 생산하는 구성효소로서의 이눌라아제)

  • Uhm, Tai-Boong;Hong, Jai-Sik;Sohn, Hee-Suk;Park, Mun-Kuk;Byun, Si-Myung
    • Applied Biological Chemistry
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    • v.28 no.3
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    • pp.131-136
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    • 1985
  • Recently, we investigated characteristics of a bacterium which has never been reported as an inulin hydrolyzing microorganism. Several properties of the isolated microorganism were aerobic, rod typed, spore forming and Gram positive. According to the Bergey's manual, this bacterium tentatively appeared to be Becillus subtilis. This bacterium produced inulase constitutively in the media containing giucose, sucrose or cellulose without inulin as a sole carbon source. Also, inulase activities of the bacterium per unit culture volume showed 1.1 unit/ml comparable to 0.9 unit/ml of Kluyveromyces fragilis with relatively short culture time.

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Biocontrol Characteristics of Bacillus Species in Suppressing Stem Rot of Grafted Cactus Caused by Bipolaris cactivora

  • Bae, Sooil;Kim, Sang Gyu;Kim, Young Ho
    • The Plant Pathology Journal
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    • v.29 no.1
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    • pp.42-51
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    • 2013
  • One of the most important limiting factors for the production of the grafted cactus in Korea is the qualitative and quantitative yield loss derived from stem rots especially caused by Bipolaris cactivora. This study is aimed to develop microbial control agents useful for the control of the bipolaris stem rot. Two bacteria (GA1-23 and GA4-4) selected out of 943 microbial isolates because of their strong antibiotic activity against B. cactivora were identified as Bacillus subtilis and B. amyloliquefaciens, respectively, by the cultural characteristics, Biolog program and 16S rRNA sequencing analyses. Both bacterial isolates significantly inhibited the conidial germination and mycelial growth of the pathogen with no significant difference between the two, of which the inhibitory efficacies varied depending on the cultural conditions such as temperature, nutritional compositions and concentrations. Light and electron microscopy of the pathogen treated with the bacterial isolates showed the inhibition of spore germination with initial malformation of germ tubes and later formation of circle-like vesicles with no hyphal growth and hyphal disruption sometimes accompanied by hyphal swellings and shrinkages adjacent to the bacteria, suggesting their antibiotic mode of antagonistic activity. Control efficacy of B. subtilis GA1-23 and B. amyloliquefaciens GA4-4 on the cactus stem rot were not as high as but comparable to that of fungicide difenoconazole when they were treated simultaneously at the time of pathogen inoculation. All of these results suggest the two bacterial isolates have a good potential to be developed as biocontrol agents for the bipolaris stem rot of the grafted cactus.

Isolation and Characterization of Lactic Acid Bacteria Producing Antimutagenic Substance from Korean Kimchi (김치로부터 항돌연변이 물질을 생산하는 유산균의 분리 및 특성)

  • Rhee, Chang-Ho;Park, Heui-Dong
    • Microbiology and Biotechnology Letters
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    • v.27 no.1
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    • pp.15-22
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    • 1999
  • Various lactic acid bacteria were isolated from Korean Kimchi in order to study their antimutagenic substances. Ames test using Salmonella typhimurium TA98 and TA100 showed the strain KLAB21 to have the highest antimutagenic activity among the 230 isolated strains against MNNG (N-methyl-N'-nitro-N-nitrosoguanidine), NPD (4-nitro-O-phenylenediamine), NQO (4-nitrosoquinoline-1-oxide) and AFB1 (aflatoxin B1). The strain was identified as Lactobacillus plantarum based on its morphological, cultural and physiological characteristics. Antimutagenic activity of L. plantarum KLAB21 was found in culture supernatant suggesting the bacterium secrete antimutagenic substance in the media. No mutagenic activity was found in the culture supernatant. The isolated strain L. plantarum KLAB21 showed much higher antimutagenic activity than L. plantarum IAM1261 which is being used industrially for fermented milk production. The antimutagenic activity of L.plantarum KLAB21 was reconfirmed by the spore-rec assay using spores of Bacillus subtilis H17($Rec^+$) and M45($Rec^-$).

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Establishment of $F_0$-value Criterion for Canned Tuna in Cottonseed Oil (참치 기름담금 통조림의 $F_0$-값 설정에 관한 연구)

  • HAN Bong-Ho;CHO Hyun-Duck;YU Hong-Sik;KIM Sang-Ho;CHUNG Youn-Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.27 no.6
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    • pp.675-681
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    • 1994
  • [ $F_0$ ]-values of the canned tuna in cottonseed oil (CTCO) were investigated under different sterilizing conditions to optimize the energy consumption and microbiological safety. The $F_0$-values were measured using a microcomputer based technique. The exact cold point was not the volumetric center of the cans, and it was located in the center of meat mass in can which had ca. $6\%$ of head space. Location of the test cans in retort showed no remarkable influence on the $F_0$-values when the cans were jumble loaded. The process time before sterilization should be shortened as much as possible to prevent the contamination of microorganisms. Thermophilic spore forming bacteria found from raw and precooked tuna were Bacillus subtilis, Bacillus cereus and Bacillus pasteurii, and the most heat resistant was Bacillus subtilis. The rational $F_0$-value for the CTCO obtained from the preservation test was regarded as 6min.

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Comparison of Urushiol Composition and Biological Activity between Fresh Sap and Fire Distilled Sap of Lacquer Tree (생칠과 화칠의 우루시올 조성 및 생리활성 비교)

  • 김명조
    • Korean Journal of Plant Resources
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    • v.11 no.1
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    • pp.40-46
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    • 1998
  • For the comparison of the urushiol composition and biological acitivity between the fresh sap and fire distilled sap of lacquer tree(Rhus vernicifera), we analysed the urushiol composition by HPLC and EI-MS, and investigated the antioxidative , antimicrobial and anticancer acitivities according to solvent fractionations. There was no difference in the urushiol composition between fresh and fire distilled saps of lacquer tree. The hexane frqctionsof two saps showed a strong DPPH radical scaverging activity (RC50 : 7.0-7.5$\mu\textrm{g}$). They also showed a strong antifungal activity onthe spore germination of Cladosporium herbarum(MIC : 8$\mu\textrm{g}$/ml), whereas they have no or low activity against the bacteria(BAcillus subtilis , Escherichia coli). In addition , hexane and butanol fractions of two saps showed a strong inhibitory activity against cultured tumour cell lines (GI50 : 0.35, 12.29$\mu\textrm{g}$/ml) in vitro. These results confirm that the fresh sap and fire distilled sap might have the similar urushiol compositions and biological activities.

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