• Title/Summary/Keyword: Bacillus strain

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Isolation and Characterization of Oligotrophic Strains with High Enzyme Activity from Buckwheat Sokseongjang (메밀 속성장 유래 효소활성 우수 저영양성 균주 분리 및 특성)

  • Lee, Sung-Young;Kim, Ji-Yeun;Baek, Sung-Yeol;Yeo, Soo-Hwan;Koo, Bon-Sung;Park, Hye-Young;Choi, Hye-Sun
    • Korean Journal of Food Science and Technology
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    • v.43 no.6
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    • pp.735-741
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    • 2011
  • Bealmijang is a short-term fermented regional product that is prepared with soybean and extra ingredients. In this study, starter strain candidates were screened from Bealmijang for fermented soybean paste products. Twenty one bacterial strains producing extracellular enzymes (amylase, cellulase, protease, xylanase and lipase) were isolated from Bealmijang, buckwheat sokseongjang. The isolates were assessed for fibrinolytic and antibacterial activities, and salt tolerance. Strain HJ18-4, identified as Bacillus subtilis (AB601598) by biochemical properties (89.6%) and 16S rDNA sequencing (100%), showed the highest enzymatic, fibrinolytic, and antibacterial activities among the isolates. Although the growth of HJ18-4 was inhibited by the increase of NaCl concentration, the growth still exceeded that of B. subtilis KACC 10114 at 5% and 10% NaCl. These results suggest that B. subtilis HJ18-4 is suitable as a starter for soybean paste manufacture.

Purification and Characterization of the Bacteriocin Produced by Lactococcus sp. KD 28 Isolated from Kimchi (김치에서 분리한 Lactococcus lactis가 생산하는 박테리오신의 정제 및 특성)

  • Lee, Ji-Young;Choi, Nack-Shick;Chun, Sung-Sik;Moon, Ja-Young;Kang, Dae-Ook
    • Journal of Life Science
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    • v.25 no.2
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    • pp.180-188
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    • 2015
  • The bacterial strain isolated from Kimchi showed antibacterial activity against Micrococcus luteus IAM 1056. The selected strain was identified as Lactococcus lactis by 16S rRNA nucleotide sequence analysis and named as Lactococcus sp. KD 28. The treatment of culture supernatant with proteinase K removed antibacterial activity, indicating its proteinaceous nature, a bacteriocin. This bacteriocin was sensitive to hydrolytic enzymes such as ${\alpha}$-chymotrypsion, trypsin, proteinase K, lipase, ${\alpha}$-amylase and subtilisin A. The bacteriocin was highly thermostable and resistant to heating at $80^{\circ}C$ for up to an hour but 50 % of the total activity was remained at $100^{\circ}C$ for 30 min. The pH range from 2.0 to 8.0 had no effect on bacteriocin activity and it was not affected by solvents such as acetonitrile, isopropanol, methanol, chloroform and acetone up to 50% concentration. The bacteriocin showed antibacterial activity against M. luteus IAM 1056, Lactobacillus delbrueckii subsp. lactis KCTC 1058, Enterococcus faecium KCTC 3095, Bacillus cereus KCTC 1013, B. subtilis KCTC 1023, Listeria ivanovii subsp. ivanovii KCTC 3444, Staphylococcus aureus subsp. aureus KCTC 1916, B. megaterium KCTC 1098 and B. sphaericus KCTC 1184. The bacteriocin was purified through ammonium sulfate concentration, SP-Sepharose chromatography and RP-HPLC. The molecular weight was estimated to be about 3.4 kDa by tricine-SDS-PAGE analysis.

Isolation and Characterization of a Paenibacillus incheonensis YK5 with Antimicrobial Activity aginst MRSA (항MRSA 활성을 보이는 Paenibacillus incheonensis YK5의 분리 및 특성)

  • Yoon, Young-Jun;Kim, Hye-Yoong;Lee, Tae-Soo;Kim, Jung-Wan
    • Korean Journal of Microbiology
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    • v.44 no.4
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    • pp.326-332
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    • 2008
  • Various bacteria were isolated from Korean soil samples based on their capability inhibiting the growth of MRSA strains. Among them, strain YK5 with the highest activity was a Gram positive sporulative bacillus with motility. It did not produce indole and no acid was formed from mannitol by the bacterium. The 16S rRNA sequence of the strain showed $95{\sim}98%$ homology with those of Paenibacillus spp.. The bacterial isolate shared the highest homology with that of P. elgii (98%), but was named as Paenibacillus incheonensis YK5 due to differences in physiological properties. Butanol extract of the P. incheonensis YK5 culture grown in SST medium at $37^{\circ}C$ for 96 hr showed a broad antimicrobial activity against Gram-positive (MRSA and Streptococcus pneumoniae) and negative (Pseudomonas aeruginosa, Salmonella spp., Shigella spp., Escherichia coli, Klebsiella pneumoniae) pathogenic bacteria and fungi (Cryptococcus neoformans and Trichophyton). The antimicrobial activity in the crude extract was stable in a broad range of temperature and pH, $20{\sim}100^{\circ}C$ and $3.0{\sim}6.0$, respectively. Therefore, the antimicrobial activity of P. incheonesis YK5 had potential as a novel antibiotics for pathogens including MRSA.

Effects of a multi-strain probiotic on growth, health, and fecal bacterial flora of neonatal dairy calves

  • Guo, Yongqing;Li, Zheng;Deng, Ming;Li, Yaokun;Liu, Guangbin;Liu, Dewu;Liu, Qihong;Liu, Qingshen;Sun, Baoli
    • Animal Bioscience
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    • v.35 no.2
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    • pp.204-216
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    • 2022
  • Objective: The aim of this study was to investigate the effects of dietary supplementation with a multi-strain probiotic (MSP) product containing of Bifidobacterium animalis, Lactobacillus casei, Streptococcus faecalis, and Bacillus cerevisiae on growth, health, and fecal bacterial composition of dairy calves during the first month of life. Methods: Forty Holstein calves (24 female and 16 male) at 2 d of age were grouped by sex and date of birth then randomly assigned to 1 of 4 treatments: milk replacer supplementation with 0 g (0MSP), 2 g (2MSP), 4 g (4MSP), and 6 g (6MSP) MSP per calf per day. Results: Supplementation of MSP did not result in any significant differences in parameters of body measurements of calves during the 30 d period. As the dosage of MSP increased, the average daily gain (p = 0.025) and total dry matter intake (p = 0.020) of calves showed a linear increase. The fecal consistency index of the 2MSP, 4MSP, and 6MSP group calves were lower than that of the 0MSP group calves (p = 0.003). As the dosage of MSP increased, the concentrations of lactate dehydrogenase (p = 0.068) and aspartate aminotransferase (p = 0.081) in serum tended to decrease, whereas the concentration of total cholesterol increased quadratically (p = 0.021). The relative abundance of Dorea in feces was lower (p = 0.011) in the 2MSP, 4MSP, and 6MSP group calves than that in the 0MSP group calves. The relative abundance of Dorea (p = 0.001), Faecalibacterium (p = 0.050), and Mitsuokella (p = 0.030) decreased linearly, whereas the relative abundance of Prevotella tended to increase linearly as the dosage of MSP increased (p = 0.058). Conclusion: The MSP product can be used to reduce the diarrhea, improve the performance, and alter the composition of the fecal bacteria in neonatal dairy calves under the commercial conditions.

Production of Maltopentaose and Biochemical Characterization of Maltopentaose-Forming Amylase

  • Kim, Young-Min;Ryu, Hwa-Ja;Lee, Sun-Ok;Seo, Eun-Seong;Lee, So-Young;Yoo, Sun-Kyun;Cho, Dong-Lyun;Kim, Do-Man;Kimura, Atsuo;Chiba, Seiya;Lee, Jin-Ha
    • Journal of Microbiology and Biotechnology
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    • v.11 no.4
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    • pp.636-643
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    • 2001
  • Bacillus sp. AIR-5, a strain from soil, produced an extracellular maltopentaose-forming amylase from amylose and soluble starch. This bacterium produced 8.9 g/l of maltopentaose from 40 g/l of soluble starch in a batch fermentation and the maltopentaose made up 90 % of the maltooligosaccharides produced (from maltose to maltoheptaose). The culture supernatant was concentrated using a 30 K molecular weight cut-off membrane and purified by DEAE-Cellulose and Sephadex G-150 column chromatographies. The purified protein showed one band on a native-PAGE and its molecular mass was estimated as 250 kDa. The 250-kDa protein was composed of tetramers of a 63-kDa protein. the isoelectric point of the purified protein was pH 6.9, and the optimum temperature for the enzyme activity was $45^{\circ}C$. The enzyme was quickly inactivated above $55^{\circ}C$, and showed a maximum activity at pH 8.5 and over 90% stability between a pH of 6 to 10. The putative N-terminal amino acid sequence of AIR-5 amylase, ATINNGTLMQYFEWYVPNDG, showed a 96% sequence similarity with that of BLA, a general liquefying amylase.

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Isolation and Characterization of a Mutant Defective in Light-activated Heterotrophic Groth from Synechocystis sp. PCC 6803 (Synechocystis sp. PCC 6803으로부터 광활성 종속영양 생장결핍 돌연변이체의 분리 및 특성)

  • Park, Mi-Seon;Lee, Young-Sook;Kim, Young-Chang
    • Korean Journal of Microbiology
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    • v.32 no.3
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    • pp.202-207
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    • 1994
  • A mutant strain PRM1 defective in light-activated heterotrophic growth was isolated from Synechocystis sp. PCC 6803. PRM1 could be grown at growth rate equivalent to Synechocystis 6803 under mixotrophic growth conditions. However, PRM1 could not be grown under light-activated heterotrophic conditions, in which a daily pulse of light for 5 min was given. These results suggest that PRM1 is not defective in heterotrophic metabolism, but in the transduction pathway of light signal essential to the growth. Plasmid patterns, absorption spectra of whole cells, and the exterior and interior structures of PRM1 were similar to those of Synechocystis 6803, except that PRM1 could not produce amorphous slime holding cells together.

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Degradation of Lignocelluloses in Rice Straw by BMC-9, a Composite Microbial System

  • Zhao, Hongyan;Yu, Hairu;Yuan, Xufeng;Piao, Renzhe;Li, Hulin;Wang, Xiaofen;Cui, Zongjun
    • Journal of Microbiology and Biotechnology
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    • v.24 no.5
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    • pp.585-591
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    • 2014
  • To evaluate the potential utility of pretreatment of raw biomass with a complex microbial system, we investigated the degradation of rice straw by BMC-9, a lignocellulose decomposition strain obtained from a biogas slurry compost environment. The degradation characteristics and corresponding changes in the bacterial community were assessed. The results showed that rapid degradation occurred from day 0 to day 9, with a peak total biomass bacterium concentration of $3.3{\times}10^8$ copies/ml on day 1. The pH of the fermentation broth declined initially and then increased, and the mass of rice straw decreased steadily. The highest concentrations of volatile fatty acid contents (0.291 mg/l lactic acid, 0.31 mg/l formic acid, 1.93 mg/l acetic acid, and 0.73 mg/l propionic acid) as well as the highest xylanse activity (1.79 U/ml) and carboxymethyl cellulase activity (0.37 U/ml) occurred on day 9. The greatest diversity among the microbial community also occurred on day 9, with the presence of bacteria belonging to Clostridium sp., Bacillus sp., and Geobacillus sp. Together, our results indicate that BMC-9 has a strong ability to rapidly degrade the lignocelluloses of rice straw under relatively inexpensive conditions, and the optimum fermentation time is 9 days.

Isolation and Identification of the Antilisterial Bifidobacterium Isolates from the Infants Fecal Samples (유아의 분변으로부터 항리스테리아 활성의 Bifidobacterium 속 균주의 분리 및 동정)

  • Kim, Song-Yi;Kim, Ki-Hwan;Youn, Soon-Yong;Yoon, Sung-Sik
    • Journal of Dairy Science and Biotechnology
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    • v.24 no.1
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    • pp.19-28
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    • 2006
  • This study was conducted to isolate antilisterial strains of the Bifidobacterium isolates from the infant feces. The bifidobacteria were isolated anaerobically on BL agar and screened for their inhibitory activity on the MRS-cysteine medium against three foodborne pathogens: Listeria monocytogenes, Bacillus cereus, and Staphylococcus aureus. Among the 52 bifidobacterial isolates, 5 strains(A24, Bl, B6, B10, and Bl2) were finally selected based on their stronger antilisterial activity against Listeria monocytogenes than other isolates tested. Morphologically, all the isolates were typically shown Y-and V-shaped under electron microscopic examination. Each isolate was primarily subjected to identification by a polymerase chain reaction(PCR) using a genus-specific primer designed for targeting the 16S rRNA gene sequence, and confirmed the primary identification data using an API-kit(Biomeriuex, France), commercially available product for identification based on biochemical and physiological traits. Of the isolates with antilisterial activity, strain A24 was finally confirmed as the Bifidobacterium longum A24.

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Characterization of the xaiF Gene Encoding a Novel Xylanase-activity- increasing Factor, XaiF

  • Cho, Ssang-Goo;Choi, Yong-Jin
    • Journal of Microbiology and Biotechnology
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    • v.8 no.4
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    • pp.378-387
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    • 1998
  • The DNA sequence immediately following the xynA gene of Bacillus stearothermophilus 236 [about l-kb region downstream from the translational termination codon (TAA) of the xynA gene]was found to have an ability to enhance the xylanase activity of the upstream xynA gene. An 849-bp ORF was identified in the downstream region, and the ORF was confirmed to encode a novel protein of 283 amino acids designated as XaiF (xylanase-activity-increasing factor). From the nucleotide sequence of the xaiF gene, the molecular mass and pI of XaiF were deduced to be 32,006 Da and 4.46, respectively. XaiF was overproduced in the E. coli cells from the cloned xaiF gene by using the T7 expression system. The transcriptional initiation site was determined by primer extension analysis and the putative promoter and ribosome binding regions were also identified. Blast search showed that the xaiF and its protein product had no homology with any gene nor any protein reported so far. Also, in B. subtilis, the xaiF trans-activated the xylanase activity at the same rate as in E. coli. In contrast, xaiF had no activating effect on the co-expressed ${\beta}-xylosidase$ of the xylA gene derived from the same strain of B. stearothermophilus. In addition, the intracellular and extracellular fractions from the E. coli cells carrying the plasmid-borne xaiF gene did not increase the isolated xylanase activity, indicating that the protein-protein interaction between XynA and XaiF was not a causative event for the xylanase activating effect of the xaiF gene.

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Investigation of Hazards from Onions and Their Cultivation Areas to Establish a Good Agricultural Practices (GAP) Model (Good agricultural practices 모델 개발을 위한 양파 및 생산 환경에서의 위해요소 조사)

  • Choi, Young-Dong;Lee, Chae-Won;Kim, Jeong-Sook;Chung, Duck-Hwa;Shim, Won-Bo
    • Korean Journal of Food Science and Technology
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    • v.45 no.6
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    • pp.785-790
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    • 2013
  • The purpose of this study was to investigate the hazards from onions and their cultivation areas. A total of 32 samples were collected from onion farms and tested for biological (sanitary indicators, and pathogenic bacteria and fungi) and chemical (heavy metals and pesticide residues) hazards. Aerobic bacteria and coliforms were detected at a level of 0.2-7.1 log CFU/g (or mL) in the soil and agricultural water, 1.6-3.6 log CFU/g on surface of the onion, 0.0-6.0 log CFU/hand (or $cm^2$) on the workers' hands, clothes, and gloves, and 4.7 log $CFU/cm^2$ on the onion bags. Fungi were detected at a level of 0.0-5.0 log CFU/g (or mL, hand, or 100 $cm^2$) in all the samples. Staphylococcus aureus was detected at a level of 1.2 log CFU/hand on the workers' hands, the detection level of Bacillus cereus was up to 4.8 log CFU/g in the soil. However, Escherichia coli (and in particular strain O157:H7), Listeria monocytogenes, and Salmonella spp. were not detected. Although heavy metals were detected in the environment (in soil and agricultural water) and pesticide residues were detected in onion, the levels were lower than the regulation limits.