• Title/Summary/Keyword: Culture Supernatant

Search Result 593, Processing Time 0.035 seconds

Xylanase Production by Bacillus sp. A-6 Isolated from Rice Bran

  • Lee, Jun-Ho;Choi, Suk-Ho
    • Journal of Microbiology and Biotechnology
    • /
    • v.16 no.12
    • /
    • pp.1856-1861
    • /
    • 2006
  • A Bacillus sp. A-6 strain that produced xylanase was isolated from rice bran. The optimal temperature and pH for xylanase activity of the culture supernatant of Bacillus sp. A-6 were 40$^{\circ}C$ and pH 7, respectively. The optimal temperature and pH for xylanase production in the xylan medium were 30$^{\circ}C$ and pH 9, respectively. The optimal concentrations of oat spelt xylan and peptone for xylanase production were 0.5% and 1.5%, respectively. The best nitrogen sources for xylanase production was beef extract, but xylanase production was also supported comparably by tryptone and peptone. The bacterial growth in the optimal xylan medium reached stationary growth phase after 12 h of incubation. The xylanase production in the culture supernatant increased dramatically during the initial 12 h exponential growth phase and then remained constant at 23.8-24.5 unit/ml during the stationary growth phase. The pH of the culture medium decreased from 8.8 to 6.7 during the exponential growth phase and subsequently increased to 8.1 during the stationary growth phase. Rice bran, sorghum bran, and wheat bran as well as oat spelt xylan induced xylanase production. The xylanase production was repressed when glucose was added to the xylan-containing medium.

Isolation and Characterization of a Thermophilic Bacillus sp. producing a Thermostable $\alpha$-glucosidase (내열성$\alpha$-glucosidase를 생산하는 호열성 Bacillus sp. 균주의 분리 및 특성)

  • 이용억
    • Journal of Life Science
    • /
    • v.8 no.4
    • /
    • pp.387-394
    • /
    • 1998
  • A thermophilic bacterium (strain DG0303) producing a thermostable $\alpha$-glucosidase was isolated from manure and identified as Bacillus sp. Strain DG0303 produced high level of $\alpha$-glucosidase compared with other thermophilic Bacillus strains. The cellular protein patterns were also compared with other Bacillus strains by sodium dodecyl sulfatepolyacrylamide gel electrophoresis(SDS-PAGE). On the basis of 16S rDNA analysis the Bacillus sp. DG0303 was found to be a member of Bacillus rDNA group 5. The optimum temperature for growth was 65$\circ$C and no growth was obtained at 40$\circ$C or 75$\circ$C. The optimum pH for growth was 5.5 to 8.5. $\alpha$-glucosidase activity was produced during growth and most activity was detected in the culture supernatant. The $\alpha$-glucosidase production was constitutive in the absence of carbohydrates. High level of enzyme activity was detected when the culture was grown on medium containing starch. Addition of glucose resulted in the repression of the $\alpha$-glucosidase production. The optimum pH and tempoerature for enzyme activity were pH 5.0 and 65$\circ$C, respectively. When analyzed by zymogram, the culture supernatant showed a single $\alpha$-glucosidase band with a molecular weight of approximately 60,000.

  • PDF

A Bacteriocin of 5-kDa in Size Secreted by Bacillus subtilis 168 (Bacillus subtilis 168 균주가 분비하는 5 kDa 크기의 Bacteriocin)

  • Kwon, Gun-Hee;Lee, Hwang-A;Kim, Jeong-Hwan
    • Microbiology and Biotechnology Letters
    • /
    • v.38 no.2
    • /
    • pp.163-167
    • /
    • 2010
  • Bacillus subtilis 168 secreted antimicrobial substance(s) into culture medium and culture supernatant inhibited growth of some Gram positive bacteria. B. cereus and Listeria monocytogenes were the most sensitive organisms. The antimicrobial activity was destroyed when culture supernatant was treated by protease and proteinase K, indicating the proteinous nature of the substance (bacteriocin). The molecular weight of the bacteriocin was estimated to be 5 kDa by Tricine SDS-PAGE. B. cereus ATCC 14579 cells were killed when exposed to the bacteriocin, indicating that the mode of inhibition was bacteriocidal. These results show that B. subtilis 168 could be useful as a starter for fermented foods such as cheonggukjang where B. cereus contamination is a major concern.

Effect of Lactococcus lactis 1370 on the Formation of Artificial Plaque (Lactococcus lactis 1370가 인공치태 형성에 미치는 영향)

  • Chung, Jin;Yim, Sung-Yee;Oh, Jong-Suk
    • The Journal of the Korean Society for Microbiology
    • /
    • v.35 no.1
    • /
    • pp.77-85
    • /
    • 2000
  • Streptococcus mutans is the most important causative bacteria of dental caries among the oral bacteria. Lactococcus lactis 1370 was isolated from the oral cavity of child. The effect of Lactococcus lactis 1370 on the formation of artificial plaque by Streptococcus mutans was studied. 1. The insoluble substances and bacteria were much more attached on the wall of disposable cuvette in the culture of Streptococcus mutans than in the combined culture of Streptococcus mutans and Lactococcus lactis 1370. 2. The mean weight of produced artificial plaque on the wires in the beaker was 131.7 mg in the culture of Streptococcus mutans only, whereas being reduced to 6.4 mg in the combined culture of Streptococcus mutans and Lactococcus lactis 1370 (p<0.05). The viable cell didn't show the significant difference between them after culturing. 3. When Streptococcus mutans was cultured in the media containing culture supernatant of Lactococcus lactis 1370 cultured in M17 broth containing 0.5% yeast extract and 5% sucrose, the mean weight of produced artificial plaque was 8.0 mg on the wires, whereas being 125.4 mg in the media without culture supernatant of Lactococcus lactis 1370 (p<0.05). The viable cell didn't show the significant difference between them after culturing. 4. When Streptococcus mutans was cultured in the media containing soluble polymer produced by Lactococcus lactis 1370, the mean weight of produced artificial plaque was significantly reduced compared with being cultured in the media without soluble polymer (p<0.05). The viable cell didn't show the significant difference between them after culturing. 5. The soluble polymer produced by Lactococcus lactis 1370 was glucan. 6. The glucan produced by Lactococcus lactis 1370 was water-soluble glucan containing ${\alpha}$-1,6-glucose linkage as the main linkage. These results suggest that the artificial plaque formed by Streptococcus mutans is inhibited by water-soluble glucan produced by Lactococcus lactis 1370.

  • PDF

Antibacterial Activities against Pathogenic Bacteria of Lactic Acid Bacteria Isolated from Allium wakegi (쪽파로부터 분리된 유산균의 병원성균에 대한 항균활성 )

  • Gil-Ha Kim;Natsag Lkhagvasuren;Batchimeg Namshir;Woan Sub Kim
    • Journal of Dairy Science and Biotechnology
    • /
    • v.41 no.3
    • /
    • pp.126-137
    • /
    • 2023
  • In this study, we isolated lactic acid bacteria from Allium wakegi and examined the usability of culture supernatants obtained from these lactic acid bacteria. The antibacterial activity of the culture supernatant obtained from the isolated lactic acid bacteria against the pathogens Escherichia and Salmonella spp. was measured. The obtained lactic acid bacteria culture medium showed significant antibacterial activity against pathogenic bacteria in a dose-dependent manner. The effects of pH and heat denaturation on the observed anti-pathogenic bacterial activity was also investigated. Adjusting the culture supernatant to pH 7 resulted in loss of all antibacterial activity against pathogenic bacteria, suggesting that the antibacterial activity of the obtained culture supernatant against pathogenic bacteria is influenced by organic acids. Assessment of the heat stability of the anti-pathogenic bacterial activity revealed that heat treatment did not diminish activity. The obtained lactic acid bacteria culture medium is thus stable against heat.

Purification and Properties of a Novel Extracellular Agarase from Marine Bacterium, Sphingomonas paucimobilis AS-1 (해양미생물 Sphingomonas paucimobilis AS-1이 생산하는 새로운 extracelluar agarase의 정제 및 특성)

  • Jung, Il-Sun;Kim, Yu-Jung;Song, Hyo-Ju;Gal, Sang-Wan;Choi, Young-Ju
    • Journal of Life Science
    • /
    • v.18 no.1
    • /
    • pp.103-108
    • /
    • 2008
  • An agar-degrading marine bacterium, strain AS-1 was isolated from the seawater. The strain AS-1 was identified as Sphingomonas paucimobilis (90% probability) by VITEK. The optimum medium for agarase activity of the isolated strain was determined to be marine medium, marine broth 2216 containing 0.1% agar as carbon source. An extracellular agarase was purified 104-fold from the culture supernatant by ammonium sulfate precipitation, ion exchange chromatography and gel filtration methods. The molecular weight of the purified enzyme was estimated to be 80 kDa by SDS-PAGE. The optimum pH and temperature for activity were 7.0 and $40^{\circ}C$, respectively. Antioxidative activity of the strain AS- was 72% in the supernatant cultured for 12 h. The culture supernatant of the strain AS-1 showed antibacterial activity against bacteria causing putrefaction and food poisoning such as Escherichia coli, Staphylococcus aureus and Proteus vulgaris. However, the cell growth of the lactic aicd forming strain, Lactobacillus plantarium was promoted by the treatment of 10% culture supernatant of an agar-degrading strain.

Antimutagenic Effects against Aflatoxin B1 on Culture Conditions of Leuconostoc mesenteroides subsp. cremoris DLAB19 Isolated from Dongchimi (동치미에서 분리한 Leuconostoc mesenteroides subsp. cremoris DLAB19의 배양조건에 따른 Aflatoxin B1에 대한 항돌연변이 효과)

  • Rhee Chang-Ho;Kim Jung-Hee;Park Heui-Dong
    • Food Science and Preservation
    • /
    • v.11 no.3
    • /
    • pp.405-410
    • /
    • 2004
  • Leuconostoc mesenteroides subsp. cremoris DLAB19 were investigated under various culture conditions to maximize the production of antimutagenic substance(s) against aflatoxin Bl(AFBl) on Salmonella enterica serovar Typhimurium TAI00 and TA98. The MRS medium containing glucose(2$\%$) as a carbon source and yeast extract(1 $\%$) as a nitrogen source resulted in the highest production of the antimutagenic substance(s) against aflatoxin Bl(AFBl) in the culture supernatant of Leu. mesenteroides subsp. cremoris DLAB19. Optimal pH of the medium, culture temperature and shaking speed for the antimutagenic substance(s) production were pH 7.0, 30$^{\circ}C$ and 150 rpm, respectively. Under the optimal condition, the antimutagenic effects of Leu. mesenteroides subsp. cremoris DLAB19 culture supernatant were 87.11 $\%$ on S. enterica serovar Typhimurium TA100 and 75.04 S. enterica serovar Typhimurium TA98.

Optimal Production and Characterization of Laccase from Fomitella fraxinea Mycelia (Fomitella fraxinea 균사체로부터 Laccase의 최적생산 및 효소적 특성)

  • Park Kyung-Mi;Park Sang-Shin
    • Microbiology and Biotechnology Letters
    • /
    • v.34 no.3
    • /
    • pp.228-234
    • /
    • 2006
  • The culture conditions were investigated to maximize the production of laccase from Fomitella fraxinea mycelia. Among the tested media, mushroom complete medium (MCM) showed the highest production of the enzyme. The optimum culture medium was 2% dextrose, 0.4% $(NH_4)_{2}HPO_4$, 0.05% $Na_{2}HPO_{4}{\cdot}7H_{2}O$, and 0.05% KCl as carbon, nitrogen, phosphorus, and inorganic salt sources respectively. SDS-PAGE followed by laccase activity staining using 2,6-djmethoxyphenol as the substrate was performed to identify the laccase activity under culture conditions studied. Zymogram analysis of the culture supernatant showed a laccase band with a molecular mass of 50 kDa. The enzyme production from F. fraxinea was reached to the highest level after the cultivation for 10 days at $25^{\circ}C$ and initial pH 8. The enzyme activity of the culture supernatant was most active at $50^{\circ}C$ and pH 5.

Overcoming Encouragement of Dragon Fruit Plant (Hylocereus undatus) against Stem Brown Spot Disease Caused by Neoscytalidium dimidiatum Using Bacillus subtilis Combined with Sodium Bicarbonate

  • Ratanaprom, Sanan;Nakkanong, Korakot;Nualsri, Charassri;Jiwanit, Palakrit;Rongsawat, Thanyakorn;Woraathakorn, Natthakorn
    • The Plant Pathology Journal
    • /
    • v.37 no.3
    • /
    • pp.205-214
    • /
    • 2021
  • The use of the supernatant from a Bacillus subtilis culture mixed with sodium bicarbonate was explored as a means of controlling stem brown spot disease in dragon fruit plants. In in vitro experiments, the B. subtilis supernatant used with sodium bicarbonate showed a strong inhibition effect on the growth of the fungus, Neoscytalidium dimidiatum, the agent causing stem brown spot disease and was notably effective in preventing fungal invasion of dragon fruit plant. This combination not only directly suppressed the growth of N. dimidiatum, but also indirectly affected the development of the disease by eliciting the dragon-fruit plant's defense response. Substantial levels of the pathogenesis-related proteins, chitinase and glucanase, and the phenylpropanoid biosynthetic pathway enzymes, peroxidase and phenyl alanine ammonia-lyase, were triggered. Significant lignin deposition was also detected in treated cladodes of injured dragon fruit plants in in vivo experiments. In summary, B. subtilis supernatant combined with sodium bicarbonate protected dragon fruit plant loss through stem brown spot disease during plant development in the field through pathogenic fungal inhibition and the induction of defense response mechanisms.

Identification and Characterization of Hydrogen Peroxide-generating Lactobacillus fermentum CS12-1

  • Kang, Dae-Kyung;Oh, H.K.;Ham, J.-S.;Kim, J.G.;Yoon, C.H.;Ahn, Y.T.;Kim, H.U.
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.18 no.1
    • /
    • pp.90-95
    • /
    • 2005
  • Lactic acid bacteria were isolated from silage, which produce high level of hydrogen peroxide in cell culture supernatant. The 16S rDNA sequences of the isolate matched perfectly with that of Lactobacillus fermentum (99.9%), examined by a 16S rDNA gene sequence analysis and similarity search using the GenBank database, thus named L. fermentum CS12-1. L. fermentum CS12-1 showed resistance to low pH and bile acid. The production of hydrogen peroxide by L. fermentum CS12-1 was confirmed by catalase treatment and high-performance liquid chromatography. L. fermentum CS12-1 accumulated hydrogen peroxide in culture broth as cells grew, and the highest concentration of hydrogen peroxide reached 3.5 mM at the late stationary growth phase. The cell-free supernatant of L. fermentum CS12-1 both before and after neutralization inhibited the growth of enterotoxigenic Escherichia coli K88 that causes diarrhea in piglets.