• 한국미생물·생명공학회지
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• 제28권5호
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• pp.243-250
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• 2000

The study was performed to investigated the excellent microbial anticaries substance which is more effective than the chlorohexidine in the dental caries treatment. For the screening of alkaliphilic microorganism, more than 1200 bacterial strains were isolated from sea soil sample. A typ-ical strain which produced the most excellent antimicrobial substance was selected. The strain was identified novel alkalophilic Bacillus sp. through the results of morphological, biochemical and chemotaxonomical characteristics and 16S rDNA sequencing and designated as Bacillus alkalophilshaggy JY-827.

• Journal of Microbiology and Biotechnology
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• 제29권12호
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• pp.1982-1992
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• 2019

The alkaliphilic, calcium carbonate precipitating Bacillus sp. strain AK13 can be utilized in concrete for self-repairing. A statistical experimental design was used to develop an economical medium for its mass cultivation and sporulation. Two types of screening experiment were first conducted to identify substrates that promote the growth of the AK13 strain: the first followed a one-factor-at-a-time factorial design and the second a two-level full factorial design. Based on these screening experiments, barley malt powder and mixed grain powder were identified as the substrates that most effectively promoted the growth of the AK13 strain from a range of 21 agricultural products and by-products. A quadratic statistical model was then constructed using a central composite design and the concentration of the two substrates was optimized. The estimated growth and sporulation of Bacillus sp. strain AK13 in the proposed medium were 3.08 ± 0.38 × 10⁸ and 1.25 ± 0.12 × 10⁸ CFU/ml, respectively, which meant that the proposed low-cost medium was approximately 45 times more effective than the commercial medium in terms of the number of cultivatable bacteria per unit price. The spores were then powdered via a spray-drying process to produce a spore powder with a spore count of 2.0 ± 0.7 × 10⁹ CFU/g. The AK13 spore powder was mixed with cement paste, yeast extract, calcium lactate, and water. The yeast extract and calcium lactate generated the highest CFU/ml for AK13 at a 0.4:0.4 ratio compared to 0.4:0.25 (the original ratio of the B4 medium) and 0.4:0.8. Twenty-eight days after the spores were mixed into the mortar, the number of vegetative cells and spores of the AK13 strain had reached 10⁶ CFU/g within the mortar. Cracks in the mortar under 0.29 mm were healed in 14 days. Calcium carbonate precipitation was observed on the crack surface. The mortar containing the spore powder was thus concluded to be effective in terms of healing micro-cracks.

• Journal of Microbiology and Biotechnology
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• 제12권1호
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• pp.18-24
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• 2002

The present study was performed to investigate the excellent microbial anticaries substance, aminoglycoside antibiotic, which is more effective than chlorhexidine for the treatment of dental caries. The aminoglycoside antibiotic against Streptococcus mutans JC-2 from a novel alkaliphilic Bacillus alkalophilshaggy JY-827 exhibited no significant difference at the treatment concentration of $2.5{\times}10^{-7}M$, however, it inhibited the activity of the Streptococcus mutans glucosyltransferase by 70.2% and 99.8% at the concentrations of $2.5{\times}10^{-7}$M\;and\;2.5{\times}10^{-6}M$, respectively. Lineweaver-Burk plot of the inhibitory aminoglycoside antibiotic showed competitive inhibition, with $K_i$ value of $6.4{\times}10^{-6}$ M. The aminoglycoside antibiotic did not show any cytotoxicity against human gingival cells. To evaluate the industrial applicability of the aminoglycoside antibiotic, a toothpaste containing this substance was prepared and tested on the extracted human teeth. The inhibitory rate of tooth calcification and calcium ion elution by the aminoglycoside antibiotic were 50% and 2.5 times, respectively. These results suggested that the aminoglycoside antibiotic from Bacillus alkalophilshaggy JY-827 is an effective agent against dental caries.

• Journal of Microbiology and Biotechnology
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• 제12권5호
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• pp.773-779
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• 2002

A gene (hap) encoding aminopeptidase from the chromosomal DNA of Bacillus licheniformis was cloned. The gene is 1,347 bp long and encodes a 449 amino acid preproprotein with a major mature region of 401 amino acids (calculated molecular mass 43,241 Da). N-Terminal sequence of the purified protein revealed a potential presence of N-terminal propeptide. The deduced primary amino acid sequence and the mass analysis of the purified protein suggested that a C-terminal peptide YSSVAQ was also cleaved off by a possible endogeneous protease. Tho amino acid sequence displayed 58% identity with that of the aminopeptidase from alkaliphilic Bacillus halodurans. This bacterial enzyme was overexpressed in recombinant Escherichia coli and Bacillus subtilis cells. Clones containing the intact hap gene, including its own promoter and signal sequence, gave rise to the synthesis of extracellular and thrmostable enzyme by B. subtilis transformants. The secreted protein exhibited the same biochemical properties and the similar apparent molecular mass as the B. lichenzyormis original enzyme.

• Journal of Microbiology and Biotechnology
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• 제15권2호
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• pp.354-361
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• 2005

Vibrio fluvialis, an enteropathogenic bacterium, produces a phospholipase which is thought to be an important factor in the pathogenesis of disease. In this study, the phospholipase gene (vfp) was identified from V fluvialis (KCTC 2473) and its sequence was determined. The entire open reading frame was composed of 1,689 nuc1eotides and 563 amino acids. The phospholipase gene (vfp) was overexpressed in Escherichia coli as a his-tag fused protein. This recombinant protein (rVFP58) was solubilized with 6 M urea and purified by Ni-NTA affinity chromatography. The action mode of rVFP58 was determined by TLC and GC-MS, and it showed phospholipase B activity, which had both phospholipase A and lysophospholipase activities. The rVFP58 showed a maximum activity at pH around 9- 10 and temperature of about 40OC, and it was stable under alkaline condition over pH 9. The cytotoxicity of rVFP58 was evaluated, using a fish cell line, CHSE-2l4, and was found to cause significant cell death after 14 h of exposure to 250 $\mu$g of the protein.

• 한국토양비료학회지
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• 제36권3호
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• pp.127-133
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• 2003

토양미생물의 생태적 특성을 평가하기 위해 화강암, 석회암 및 현무암을 모재로 한 12개의 토양통의 미생물적 특성을 조사하였다. 토양미생물상은 토양화학성과 유의한 상관관계를 보였는데, 특히 토양 pH는 형광성 슈도모나스와, 토양 EC는 방선균, 사상균, 중온성 Bacillus 및 호알카리성균과 정의 관계를 보였다. 포자형성균인 Bacillus는 화강암과 현무암에 비해 석회암토양에서 높은 균수를 보였으나, 그람음성균은 모암간에 비슷한 수준을 유지하였다. 화강암유래 토양에서는 Burkholderia, Pseudomonas, Ralstonia가, 석회암토양에서는 Pseudomonas, Burkholderia, Phyllobacterium가, 현무암토양에서는 Burkholderia속이 우점적으로 분포하고 있었다.

• Journal of Microbiology and Biotechnology
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• 제23권5호
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• pp.661-667
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• 2013

Lipase-producing bacterial strains were isolated from Antarctic soil samples using the tricaprylin agar plate method. Seven strains with relatively strong lipase activities were selected. All of them turned out to be Bacillus pumilus strains by the 16S rRNA gene sequence analysis. Their corresponding lipase genes were cloned, sequenced, and compared. Finally, three different Bacillus pumilus lipases (BPL1, BPL2, and BPL3) were chosen. Their amino acid sequence identities were in the range of 92-98% with the previous Bacillus pumilus lipases. Their optimum temperatures and pHs were measured to be $40^{\circ}C$ and pH 9. Lipase BPL1 and lipase BPL2 were stable up to $30^{\circ}C$, whereas lipase BPL3 was stable up to $20^{\circ}C$. Lipase BPL2 was stable within a pH range of 6-10, whereas lipase BPL1 and lipase BPL3 were stable within a pH range of 5-11, showing strong alkaline tolerance. All these lipases exhibited high hydrolytic activity toward p-nitrophenyl caprylate ($C_8$). In addition, lipase BPL1 showed high hydrolytic activity toward tributyrin, whereas lipase BPL2 and lipase BPL3 hydrolyzed tricaprylin and castor oil preferentially. These results demonstrated that the three Antarctic Bacillus lipases were alkaliphilic and had a substrate preference toward short- and medium-chain triglycerides. These Antarctic Bacillus lipases might be used in detergent and food industries.

• Journal of Microbiology and Biotechnology
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• 제17권9호
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• pp.1483-1490
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• 2007

A Gram-negative, motile by tuft flagella, obligately aerobic chemoorganoheterotrophic, sphere-form bacterium, designated $IMCC3135^T$, was isolated from the Antarctic surface seawater of King George Island, West Antarctica. The strain was mesophilic, neutrophilic, and requiring NaCl for growth, but neither halophilic nor halotolerant. The 16S rRNA gene sequence analysis indicated that the strain was most closely related to genera of the order Chromatiales in the class Gammaproteobacteria. The most closely related genera showed less than 90% 16S rRNA gene sequence similarity and included Thioalkalispira (89.9%), Thioalkalivibrio (88.0%-89.5%), Ectothiorhodospira (87.9%-89.3%), Chromatium (88.3%-88.9%), and Lamprocystis (87.7%-88.9%), which represent three different families of the order Chromatiales. Phylogenetic analyses showed that this Antarctic strain represented a distinct phylogenetic lineage in the order Chromatiales and could not be assigned to any of the defined families in the order. Phenotypic characteristics, including primarily non-phototrophic, non-alkaliphilic, non-halophilic, and obligately aerobic chemoheterotrophic properties, differentiated the strain from other related genera. The very low sequence similarities (<90%) and distant relationships between the strain and members of the order suggested that the strain merited classification as a novel genus within a novel family in the order Chromatiales. On the basis of these taxonomic traits, a novel genus and species is proposed, Granulosicoccus antarcticus gen. nov., sp. nov., in a new family Granulosicoccaceae fam. nov. Strain $IMCC3135^T\;(=KCCM42676^T=NBRC\;102684^T)$ is the type strain of Granulosicoccus antarcticus.

• 한국토양비료학회지
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• 제35권4호
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• pp.243-252
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• 2002

토양생태계는 정량화하기 어려운 다양한 규칙이 다중적으로 조화되어 끊임없이 변하고있어 어떤 현상을 획일적으로 평가하기 어렵다. 그러나, 미생물은 환경에 대한 감수성이 빨라 토양의 생물적인 특성 변화를 가장 먼저 나타내기 때문에, 미생물은 생태계의 가변적 변동상의 지표가 될 수 있다. 또한 토양에 생명력을 부여해주는 기능을 가지고 있어 토양미생물의 작용 및 기능 평가는 토양 건전성 관리에 있어 중요한 역할을 한다. 따라서 본 연구는 생물학적 토양 건전성 평가 지표요인 개발하고자 주요 모재별 미경지 토양의 생물학적 토양특성을 평가하고자 수행하였다. 주요 모재별 미경지 토양의 미생물 밀도는 현무암유래 토양에서 낮고 석회암유래 토양에서 높은 경향이었으나, 그람 음성균은 비슷한 수준을 유지하고 있었다. 토양 건조처리시 미생물의 생존율은 형광성 Pseudomonas속이 0.1%로 가장 낮았으나 고온성 Bacillus속과 호알카리성균은 90%이상으로 높았다. 토양미생물체량과 미생물 유fo 유기탄소량 간에는 정의 상관관계를 나타냈으며, 토양 건조처리 효과는 토양별 9.7~95.0% 범위로 평균 39.8%였다. 건토를 재습윤하였을 때 중온성 Bacillus속은 원래 균수보다 높게 복원되는 토양이 많았다. 토양 pH는 미생물수 복원율에 정의 영향을 주었다. 건토의 재습윤화에 의한 토양 미생물체량 복원율은 평균 65.3%였으며, 재습윤토의 미생물체 복원량과 형광성 Pseudomonas속간에 유의한 정의 상관관계가 있었다.