• Title/Summary/Keyword: Thermophilic Bacillus

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Analysis of the Structure of the Bacterial Community in the Livestock Manure-based Composting Process

  • Sasaki, Hiraku;Nonaka, Jun;Otawa, Kenichi;Kitazume, Osamu;Asano, Ryoki;Sasaki, Takako;Nakai, Yutaka
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.1
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    • pp.113-118
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    • 2009
  • We investigated the structure of bacterial communities present in livestock manure-based composting processes and evaluated the bacterial succession during the composting processes. Compost samples were derived separately from swine manure, dairy manure and sewage sludge. The structure of the bacterial community was analyzed by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) using universal eubacterial primers. The genus Bacillus and related genera were mainly detected following the thermophilic composting phase of swine and dairy manure composts, and the members of the phylum Bacteroidetes were mainly detected in the cattle manure waste-based and sewage sludge compost. We recovered and sequenced limited number of the bands; however, the PCR-DGGE analysis showed that predominant diversities during the composting processes were markedly changed. Although PCR-DGGE analysis revealed the presence of different phyla in the early stages of composting, the members of the phylum Firmicutes and Bacteroidetes were observed to be one of the predominant phyla after the thermophilic phase.

Asymmetric Sythesis of Unnatural L-Amino Acids Using Thermophilic Aromatic L-Amino Acid Transaminase

  • Cho, Byung-Kwan;Seo, Joo-Hyun;Kim, Ju-Han;Lee, Chang-Soo;Kim, Byung-Gee
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.4
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    • pp.299-305
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    • 2006
  • Aromatic L-amino acid transaminase is an enzyme that is able to transfer the amino group from L-glutamate to unnatural aromatic ${\alpha}-keto$ acids to generate ${\alpha}-ketoglutarate$ and unnatural aromatic L-amino acids, respectively. Enrichment culture was used to isolate thermophilic Bacillus sp. T30 expressing this enzyme for use in the synthesis of unnatural L-amino acids. The asymmetric syntheses of L-homophenylalanine and L-phenylglycine resulted in conversion yields of >95% and >93% from 150 mM 2-oxo-4-phenylbutyrate and phenylglyoxylate, respectively, using L-glutamate as an amino donor at $60^{\circ}C$. Synthesized L-homophenylalanine and L-phenylglycine were optically pure (>99% enantiomeric excess) and continuously pre-cipitated in the reaction solution due to their low solubility at the given reaction pH. While the solubility of the ${\alpha}-keto$ acid substrates is dependent on temperature, the solubility of the unnatural L-amino acid products is dependent on the reaction pH. As the solubility difference between substrate and product at the given reaction pH is therefore larger at higher temperature, the thermophilic transaminase was successfully used to shift the reaction equilibrium toward rapid product formation.

Medium characteristics during the outdoor-composting stage of medium preparation with a prototype medium turner in button mushroom cultivation (양송이 배지교반기 시제품을 활용한 배지 조제시 야외 발효단계별 배지의 특성)

  • Lee, Chan-Jung;Yu, Byeong-Kee;Lee, Eun-Ji;Park, Hae-Sung;Kong, Won-Sik;Kim, Yeong-Ho
    • Journal of Mushroom
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    • v.16 no.1
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    • pp.45-50
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    • 2018
  • This study was performed to compare medium characteristics during the composting stage for medium turning performed using an excavator agitator and a prototype medium turner in button mushroom cultivation. The changes in temperature in the medium did not significantly differ between the treatments until the 3rd turn performed using the excavator agitator. However, during the 4th and 5th turns, the temperature increased during turning with the prototype medium turner. During outdoor composting, various types of microorganisms such as thermophilic bacteria (Bacillus spp.), Actinomycetes, fluorescent Pseudomonas spp., and filamentous fungi were found to be distributed in the medium. The counts of aerobic bacteria and fluorescent Pseudomonas spp. did not significantly differ between treatments, and the counts of thermophilic bacteria and thermophilic actinomycetes were slightly higher during turning with the prototype medium turner. The rice straw was slightly shorter and water content lower for the prototype medium turner. There was no significant difference between pH and EC treatments. The L, a, and b values tended to increase on turning with the prototype medium turner.

Bacterial Community Dynamics during Composting of Food Wastes (음식물 쓰레기 퇴비화 과정에 따른 세균군집 구조의 변화)

  • Shin, Ji-Hye;Lee, Jin-Woo;Nam, Ji-Hyun;Park, Se-Yong;Lee, Dong-Hun
    • Korean Journal of Microbiology
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    • v.45 no.2
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    • pp.148-154
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    • 2009
  • Composting is a biological process converting solid organic waste into valuable materials such as fertilizer. The change of bacterial populations in a composting reactor of food waste was investigated for 2 months. Based on shifts in temperature profile, the composting process could be divided into the first phase ($2^{\circ}C\sim55^{\circ}C$), the second phase ($55^{\circ}C\sim97^{\circ}C$), and the third phase ($50^{\circ}C\sim89^{\circ}C$). The number of total bacteria was $1.66\times10^{11}$ cell/g, $0.29\times10^{11}$ cell/g, and $0.28\times10^{11}$ cell/g in the first, second, and third stages, respectively. The proportions of thermophiles increased from 33% to 89% in the second stage. T-RFLP analysis and nucleotide sequencing of 16S rRNA gene demonstrated that the change of bacterial community structure was coupled with shifts in composting stages. The structure of bacterial community in the ultra-thermophilic second stage reflected that of seeding starter. The major decomposers driving the ultra-thermophilic composting were identified as phylotypes related to Bacillus and Pseudomonas.

Molecular Cloning and Expression of an Endo-xylanase Gene from Bacillus stearothermuphilus into Escherichia coli (Bacillus stearothermophilus로 부터 Endo-xylanase 유전자의 클로닝 및 Escherichia coli에서의 발현)

  • 조상구;박성수;박영인;최용진
    • Microbiology and Biotechnology Letters
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    • v.20 no.3
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    • pp.271-279
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    • 1992
  • Genomic DNA of Bacillus stearothemzophilus, which expressed alkalophilic and thermophilic xylanases, was partially digested with HindIII, cloned into pBR322, and subsequently transferred into the Escherichra coli HB101 cells. Three among 5, 000 transformants screened formed clear zones around their colonies. From the functional clones, three recombinant plasmids (pMG11, pMG12 and pMG13) had been isolated, and they were identified to carry the same 4 kb HindIII fragment originated from B. stearothemzophilus which was responsible for the xylanase activity. pMGl3, however, had the foreign DNA of opposite orientation compared to the other two recombinant plasmids. This recombinant plasmid gave much lower xylanase activity. B. stearothermophilus was observed to produce at least three xylanase activities as evidenced by the PAGE-xylan zymogram. The xylanase from E. coli HB101/pMG12 was judged to correspond to the largest among the three B. stearothermophilus xylanases observed in the zymogrom. The enzyme hydrolyzed xylooligosaccharides larger than xylotriose and degraded xylan to produce xylobiose and xylotriose as major products. The xylanase was considered to have trans-xylosidase activity, too.

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A Thermostable Protease Produced from Bacillus sp. DF 218 (Bacillus sp. DF218이 생산하는 내열성 단백질 분해효소)

  • Lee, Joung-Hee;Bai, Dong-Hoon
    • Korean Journal of Food Science and Technology
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    • v.36 no.1
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    • pp.105-110
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    • 2004
  • Microorganism (strain DF 218) producing thermostable pretense was isolated from Korean soil and compost. It was Gram-positive, rod-shaped, aerobic, and spore-forming with yellowish white colony color, Temperature range for growth at pH 6.5 was $30-65^{\circ}C$, with optimum growth at $60^{\circ}C$. pH range for growth at $60^{\circ}C$ was 5-7 with optimum of 6.5, which indicates strain DF 218 to be thermophilic. The 16S rDNA sequence of strain DF 218 had 95% sequence similarity with that of Bacillus flexus. Based on physiological properties and phylogenetic analysis, we proposed the isolated strain as Bacillus sp. DF 218. Pretense was produced aerobically at $60^{\circ}C$ for 32 hr in a medium (pH 6.5) containing 1% each trypton, glucose, and NaCl. Its molecular weight was estimated as 61 kDa, with optimum temperature and pH of $60^{\circ}C$ and 7.5, respectively.

Studies on the Production of Thermostable Amylase. Part 1. Optimal Culture Conditions and Purification of Enzyme. (내열성 Amylase의 생산에 관한 연구 (제1보) 최적배양조건과 효소의 정제)

  • 오두환;이강표;변유량;유주현
    • Microbiology and Biotechnology Letters
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    • v.9 no.2
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    • pp.91-97
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    • 1981
  • A thermophilic soil isolate Bacillus sp. Y-127 was selected for the production of thermostable amylase. The strain was used for the enzyme production and the thermostable amylase was characterized. The optimum cultural conditions for the enzyme production were 6$0^{\circ}C$ at pH 7.0 for 32 hours using a mineral medium containing 2% soluble starch and 0.2% yeast extract. The extra-cellular enzyme was purified about 123-folds with about 6% recovery. The purified enzyme was stable at pH between 4.0 and 7.0, and temperature up to 6$0^{\circ}C$.

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금속 이온을 이용한 Bacillus Stearothermophilus 호열성 단백질 분해효소의 역가 향상 및 호열 ${\cdot}$ 호기성 소화공정에의 응용

  • Kim, Yeong-Gi;Bae, Jin-Hye;Lee, Won-Hong;Choe, Jeong-U
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.167-170
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    • 2000
  • Proteolytic hydrolysis is one of the main enzymatic reaction of waste activated sludge (WAS) digestion. Pretense excreted from Bacillus stearothermophilus (ATCC 31197) showed optimum temperature of $75^{\circ}C$ for maxium heat stable proteolytic activity against azo casein. The dependency of $Ca^{2+}$, $Zn^{2+}$ on heat stability of proteolytic enzymes were measured with various concentrations. It was shown that $Ca^{2+}$ ion enhanced heat stability of these enzymes. Then thermophilic aerobic digestion (TAD) was performed using B. sterothermophilus with the addition of divalent ions. Performance of TAD process with ATCC 31197 activated by $Ca^{2+}$, $Zn^{2+}$ions in terms of dissolved organic carbon (DOC) concentration, extracellular protein concentration, and scanning electrion microscopy (SEM) analysis. The best result of protein reduction concentration in digestion test was obtained with the addition of 2 mM $Ca^{2+}$ ion.

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Study of Thermostable Chitinase Enzymes from Indonesian Bacillus K29-14

    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.647-652
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    • 2004
  • Thermophilic microorganisms capable of producing chitinase enzymes were screened from samples collected from several crater and geothermal areas. The chitinolytic microorganisms were grown in a selective medium containing colloidal chitin. The Bacillus K29-14 isolate was found to exhibit the highest chitinase and chitin deacetylase activities. When grown in a chitin-containing medium, the isolate produced extracellular chitinase after 24 h of incubation. The optimum temperature and pH for the chitinase were $55^\circ{C}$ and pH 7, respectively, while those for the chitin deacetylase were $55^\circ{C}$ and pH 8, respectively. The thermostable chitinase and chitin deacetylase also retained 80- 90% of their activity after incubation for 5 h at $70^\circ{C}$. The divalent cations $CoCl_2\;and\;NiCl_2$, increased the chitinase activity, while $ZnCl_2$, inhibited the enzyme. The chitin deacetylase was also activated by the presence of $MgCl_2$ and inhibited by $MnCl_2,\;NiCl_2,\;and\;CaCl_2$. A zymogram analysis revealed several forms of chitinase, with a 67 kDa form being the major enzyme.

PCR-DGGE Analysis of the Microbial Communities in Three Different Chinese "Baiyunbian" Liquor Fermentation Starters

  • Xiong, Xiaomao;Hu, Yuanliang;Yan, Nanfeng;Huang, Yingna;Peng, Nan;Liang, Yunxiang;Zhao, Shumiao
    • Journal of Microbiology and Biotechnology
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    • v.24 no.8
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    • pp.1088-1095
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    • 2014
  • A systematic investigation was performed on the bacterial, Bacillus, fungal, and yeast communities of the three types of Daqu (mechanically prepared, manually prepared, and mixed prepared) used in Baiyunbian Company by reconditioning PCR-denaturing gradient gel electrophoresis (PCR-DGGE). The DGGE results showed that the microbes in the three types of Daqu were mainly thermotolerant and thermophilic microbes, and the most dominant bacterial species were Bacillus and Virgibacillus, followed by Lactobacillus and Trichococcus. Furthermore, the dominant fungi were found to be molds, such as Rasamsonia, Penicillium, Aspergillus, and Monascus, and the dominant yeasts were Saccharomyces cerevisiae, Saccharomycopsis fibuligera, Pichia anomala, and Debaryomyces hansenii. In general, the three types of Daqu showed slight differences in microbial communities, and the Shannon indexes (H') of the manually prepared and mechanically prepared Daqu were similar. The results suggest that mechanically prepared Daqu can replace manually prepared Daqu in liquor production, and this research provides useful information for liquor production and process improvement.