• Title/Summary/Keyword: Bacillus sp. enrichment culture

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Purification and Characterization of Biosurfactant from Bacillus sp. DYL130 (Bacillus sp. DYL130 균주의 Biosurfactant의 정제 및 특성)

  • Park, In-Hye;Kim, Sun-Hee;Lee, Sang-Cheol;Ha, Soon-Ok;Lee, Yong-Seok;Ryu, Ah-Reum;Kim, Keun-Ki;Choi, Yong-Lark
    • Journal of Marine Bioscience and Biotechnology
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    • v.1 no.4
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    • pp.268-274
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    • 2006
  • Bacillus sp. DYL130 producing biosurfactant was isolated from soil samples in the Duck-yu mountain and identified as Bacillus sp. by analysis of 16S rDNA sequence. Purification of the biosurfactant was performed by using affinity chromatography and TLC. The biosurfactant of culture medium from Bacillus sp. DYL130 was eluted with 100% methanol using affinity chromatography. To remove methanol, a rotary evaporator was used and enrichment sample was dissolved in alkaline water(pH 10). The purified biosurfactant was identified by TLC. It was confirmed that the Rf value of the biosurfactant was 0.78. Antifungal activity against Botrytis cineria was showed the strongly activity as active antagonist. Maximum emulsification activity and stability were obtained from soybean oil. The critical micelle concentration (CMC) of purified biosurfactant was 35mg/l and the purified biosurfactant inhibited biofilm forming by Bacillus sp..

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Thermostable $\alpha$-Amylase Production by Thermophilic Bacillus sp. TR-25 lsolated from Extreme Enviroment (극한환경에서 분리한 고온성 Bacillus sp. TR-25에 위한 내열성 $\alpha$-amylase의 생산)

  • 노석범;손홍주;이종근
    • Journal of Life Science
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    • v.7 no.1
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    • pp.30-38
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    • 1997
  • For screening thermostable $\alpha$-amylase from thermophiles, various samples from extreme environments such as hot spring and sewage near them, and compoat, wereexamined microbial growth in enrichment culture medium at 55$\circ$C on the assumption that enzymes from thermophiles are inevitable thermostable. One strain showing higher $\alpha$-amylase activity was pure cultured and designated as Bacillus sp. TR-25 from the results of morphological, cultural and physiological characteristics. The most important carbon sourses for the enzyme production were soluble starch, dextrin, potato starch and corn starch. Glucose and fructose had a catabolite repression on the enzyme production. The good nitrogen sources for the enzyme production were yeat extract, nutrient broth, tryptone, corn steep liquor and ammonium sulfate. The enzyme production was accelerated by addition of CaCl$_{2}$. $\cdot $ H$_{2}$O. The optimal medium composition for the enzyme production was soluble starch 2.0%, yeast extract 0.55, CaCl$_{2}$ $\cdot $ 2H$_{2}$O 0.015, Tween 80 0.001%, pH8.0, respectively. In jar fermenter culture, this strain shows a rapid growth and required cheaper carbon and nitrogen source. These properties are very useful to fermentation industry. The $\alpha$-amylase of this strain demonstrated a maximum activity at 80$\circ$C, pH 5.0, respectively. And calcium ion did not improve thermostability of the enzyme. At 10$0^{\circ}C$, this enzyme has 235 of relative activity. Transformation was carried out by thermophilic Bacillus sp. TR-25 genomic DNA. As a result, the transformant has increased thermostable $\alpha$-amylase activity.

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점토로부터 철불순물의 생물학적 제거에 미치는 탄소원의 영향

  • Lee, Eun-Young;Cho, Kyung-Suk;Ryu, Hee-Wook;Bae, Moo
    • Microbiology and Biotechnology Letters
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    • v.25 no.6
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    • pp.552-559
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    • 1997
  • Fe (III) impurities in clay could be microbially removed by inhabitant dissimilatory Fe (III) reducing microorganisms. Insoluble Fe (III) in clay particles was leached out as soluble reductive form, Fe (II). The microorganisms removed from 10 to 45% of the initial Fe (III) when each sugar was supplemented to be in ranges of 1 - 5 % (w/w; sugar/clay). The microorganisms reduced 2.1 - 12.8 mol of Fe (III) per 100 mol of carbon in sugars metabolized when sugars such as glucose, maltose, and sucrose were used as sole carbon source. Bacillus sp. IRB-W and Pseudomonas sp. IRB-Y were isolated from the enrichment culture of the clay. The isolates were considered to participate in metabolizing organic compounds to fermentative intermediates with relatively little Fe (III) reduction at initial Fe (III) reduction process. By the microbial treatment, the whiteness of the clay was increased form 63.20 to 79.64, whereas the redness was obviously decreased form 13.47 to 3.55. This treatment did not cause any unfavorable modifications in mineralogical compositions of the clay.

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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.

Isolation and Characterization of Chlorothalonil-dissipating Bacteria from Soil. (토양으로부터 Chlorothalonil 전환 미생물의 분리 및 특성)

  • 이수현;신재호;최준호;박종우;김장억;이인구
    • Microbiology and Biotechnology Letters
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    • v.32 no.1
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    • pp.96-100
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    • 2004
  • Chlorothalonil is a wide-spectrum fungicide that is widely used in the world. Chlorothalonil is known as a potential toxic pollutant due to its high application rate, persistence, and toxicity to humans and other species. With the Increase of necessity of bioremediation, this study was conducted to isolate the chlorothalonil dissipation bacteria from soil. Soil samples were collected from 184 sites of farmland and wastewater disposal soil.661 strains resistant to chlorothalonil were isolated by dilution method from chlorothalonil-containing enrichment culture. After incubating at $30^{\circ}C$ in 1/10 LB media containing 10 ppm of chlorothalonil for a week, dissipation ability of chlorothalonil was investigated by HPLC. Finally, a strain SH35B, capable of dissipating chlorothalonil efficiently, was selected. The strain SH35B was identified as Ochrobactrum sp. Ten ppm of chlorothalonil In 1/10 LB media were completely dissipated by the growth of Ochrobactrum sp. SH35B for 30 h at $30^{\circ}C$. In the isolated strain, the content of glutathione and the activity of glutathione S-transferase were supposed to be ones of the Important factors for chlorothalonil dissipation and were higher than those of control strains, Escherichia coli and Bacillus subtilis.

A Study on Microbial Community and Microbial Degradation of Diesel (디젤의 미생물 분해와 군집에 관한 연구)

  • Choi, Hee-Chol;Cho, Yoon-A;Choi, Sang-Il;Lee, Tae-Jin
    • Journal of Korean Society of Environmental Engineers
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    • v.32 no.5
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    • pp.509-516
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    • 2010
  • This study investigates characteristics of diesel degradation and variations of microbial community with the soil enrichment cultures. The cultures has yellow(YE-5) and transparent color's(WH-5) colony on solid plate medium. The bacillus type of YE-5 and WH-5 cultures showed diesel degradation at the rate of 99.07mg-Diesel/$L{\cdot}day$ and 57.82mg-Diesel/$L{\cdot}day$ in the presence of 1%(v/v) initial diesel concentration. Diesel degradation was 1.7 times faster than WH-5 culture. YE-5 or WH-5 culture could degrade a wide range of diesel compounds from $C_8$ to $C_24$. Microbial community analysis by PCR-DGGE technique shows that Psedomonas, Klebsiella, Escherichia and Stenotrophomonas as proteobacteria take role on the diesel degradation. uncultured Senotrophomonas sp. was only detected with YE-5 culture. It is concluded that proper combination of the microorganism should be present to stimulate the degradation of diesel and further studies are recommended for the effect of uncultured Senotrophomonas sp. or Escherichia hermannii on diesel degradation.