• Title/Summary/Keyword: Activity Sequencing

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Properties of Antimicrobial Substances Produced by Bacillus amyloliquefaciens CJW15 and Bacillus amyloliquefaciens SSD8 (Bacillus amyloliquefaciens CJW15와 SSD8이 만드는 항균물질들의 특성)

  • Liu, Xiaoming;Shim, Jae Min;Yao, Zhuang;Lee, Jae Yong;Lee, Kang Wook;Kim, Hyun-Jin;Ham, Kyung-Sik;Kim, Jeong Hwan
    • Microbiology and Biotechnology Letters
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    • v.44 no.1
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    • pp.9-18
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    • 2016
  • Two Bacillus strains, CJW15 and SSD8, with strong antibacterial activities were isolated from cheonggukjang. Both were identified as B. amyloliquefaciens strains after gene sequencing of rRNA and recA. CJW15 strongly inhibited the growth of B. cereus (ATCC14579), Listeria monocytogenes (ATCC19111), and Lactococcus lactis (ATCC11454). In comparison, SSD8 inhibited the growth of B. cereus (ATCC14579) and Enterococcus faecium (ATCC19953). The antibacterial activities of the two strains were not affected when exposed to a temperature of $100^{\circ}C$ for 15 min and were quite stable in acidic (pH 3) and alkaline (pH 12) pH conditions. Enzymatic treatments (trypsin, pepsin, proteinase K, and protease) had no effect on the activity of CJW15, but reduced the activity of SSD8 by half. Both isolates possess genes involved in the synthesis of lipopeptides (e.g. surfactin, fengycin, iturin, and iturin A), and genes encoding subtilin, a bacteriocin. Moreover, both isolates have fibrinolytic activities as well.

Nucleotide Sequence, Structural Investigation and Homology Modeling Studies of a Ca2+-independent α-amylase with Acidic pH-profile

  • Sajedi, Reza Hassan;Taghdir, Majid;Naderi-Manesh, Hossein;Khajeh, Khosro;Ranjbar, Bijan
    • BMB Reports
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    • v.40 no.3
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    • pp.315-324
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    • 2007
  • The novel $\alpha$-amylase purified from locally isolated strain, Bacillus sp. KR-8104, (KRA) (Enzyme Microb Technol; 2005; 36: 666-671) is active in a wide range of pH. The enzyme maximum activity is at pH 4.0 and it retains 90% of activity at pH 3.5. The irreversible thermoinactivation patterns of KRA and the enzyme activity are not changed in the presence and absence of $Ca^{2+}$ and EDTA. Therefore, KRA acts as a $Ca^{2+}$-independent enzyme. Based on circular dichroism (CD) data from thermal unfolding of the enzyme recorded at 222 nm, addition of $Ca^{2+}$ and EDTA similar to its irreversible thermoinactivation, does not influence the thermal denaturation of the enzyme and its Tm. The amino acid sequence of KRA was obtained from the nucleotide sequencing of PCR products of encoding gene. The deduced amino acid sequence of the enzyme revealed a very high sequence homology to Bacillus amyloliquefaciens (BAA) (85% identity, 90% similarity) and Bacillus licheniformis $\alpha$-amylases (BLA) (81% identity, 88% similarity). To elucidate and understand these characteristics of the $\alpha$-amylase, a model of 3D structure of KRA was constructed using the crystal structure of the mutant of BLA as the platform and refined with a molecular dynamics (MD) simulation program. Interestingly enough, there is only one amino acid substitution for KRA in comparison with BLA and BAA in the region involved in the calcium-binding sites. On the other hand, there are many amino acid differences between BLA and KRA at the interface of A and B domains and around the metal triad and active site area. These alterations could have a role in stabilizing the native structure of the loop in the active site cleft and maintenance and stabilization of the putative metal triad-binding site. The amino acid differences at the active site cleft and around the catalytic residues might affect their pKa values and consequently shift its pH profile. In addition, the intrinsic fluorescence intensity of the enzyme at 350 nm does not show considerable change at pH 3.5-7.0.

Studies on the Comparative Analysis of Mating Locus (Y-region) of Schizophyllum commune (치마버섯 Mating Locus(Y-region)의 비교분석에 관한 연구)

  • 이인선;박동철
    • Journal of Life Science
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    • v.12 no.2
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    • pp.173-181
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    • 2002
  • This study was conducted to do the comparative analysis of mating type locus controlling the direct formation of fruiting body in Schizophyllum commune which is indigenous to North America with that of other identified mating locus. The 3120 bp Y-region nucleotide of A $\alpha$ 3 mating locus activating a developmental pathway in S. commune was determined, and appeared to have about 96% homology to S. commune 1-71 $A\alpha$3 allele indigenous to South America, showing strongly a conservative feature. This nucleotide analysis also showed above 96% homology highly in the seven presumed exons, and about 97% in the acidic rich region (AR), about 99% in homeodomain (H7), about 97% in the basic rich region (BR), about 95% in the serine rich region (Ser) respectively. In the comparative analysis to the translated polypeptide sequence, S. commune A $\alpha$ 3 mating locus containing Y-region also showed about 97% homology to the region of S. commune indigenous to North America, but the identity ratio to Y1 including Y4, Y5, Y6 different allele types was declined to about 41~49%. In the analysis of functional loci controlling mating activity, it is assumed to have a highly conservative feature showing about 98% homology in homeodomain polypeptide. Especially, it is notable that the homology ratio of above 85% in homeodomain motif between mating type alleles was higher than in the AR, BR, Ser showing about 10~50% homology.

Plant Growth-Promoting Activity and Genetic Diversity of Endophytic Fungi Isolated from Native Plants in Dokdo Islands for Restoration of a Coastal Ecosystem (해안 생태계의 복원을 위하여 독도에 자생하는 식물로부터 분리된 내생진균류의 식물생장촉진활성과 유전학적 다양성)

  • You, Young-Hyun;Yoon, Hyeokjun;Kim, Hyun;Lim, Sung Hwan;Shin, Jae-Ho;Lee, In-Jung;Choo, Yeon-Sik;Kim, Jong-Guk
    • Journal of Life Science
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    • v.23 no.1
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    • pp.95-101
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    • 2013
  • In this study, plant samples of five species were collected from the Dokdo islands in South Korea. Plant samples such as Asparagus schoberioides, Corydalis platycarpa, Festuca rubra, Sedum oryzifolium, and Setaria viridis were collected from the Dongdo and Seodo. Endophytic fungal strains were isolated from the roots of five plants from the Dokdo islands. Thirty-three fungal strains were isolated from these native plants. All the endophytic fungi were analyzed by internal transcribed spacer (ITS) sequencing (ITS containing ITS1, 5.8s, and the ITS2 region). Waito-c rice seedlings were treated with fungal culture filtrates to test their plant growth-promoting activity. A bioassay of the D-So-1-1 fungal strain isolated from S. oryzifolium confirmed that it has the highest plant growth-promoting activity. All the endophytic fungi belong to four orders: Eurotiales (86%), Capnodiales (3%), Hypocreales (4%), and Incertae sedis (7%). The endophytic fungi were classified as Ascomycota, which contained Aspergillus (12%), Cladosporium (3%), Eurotium (3%), Fusarium (18%), Microsphaeropsis (6%), and Penicillium (58%) at the genus level.

Antibacterial Activity against Food-poisoning Causing Bacteria and Characterization of Lactobacillus plantarum YK-9 Isolated from Kimchi (김치에서 분리한 세균인 Lactobacillus plantarum YK-9의 식중독 원인세균에 대한 항균활성 및 특성)

  • Song, You-Jin;Park, Su-Ho;You, Ji-Young;Cho, Yun-Seok;Oh, Kye-Heon
    • KSBB Journal
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    • v.24 no.3
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    • pp.273-278
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    • 2009
  • The purpose of this work was to investigate the antibacterial activity of Lactobacillus plantarum YK-9 isolated from fermented Kimchi. Morphological and biochemical characteristics of L. plantarum YK-9 were examined. Phylogenetic analysis using 16S rRNA sequencing was performed to identify the strain, and the strain could be assigned to Lactobacillus plantarum, designated as L. plantarum YK-9. The strain was registered in GenBank as [FJ669130]. During the incubation period of L. plantarum YK-9, the changes of bacterial growth and residual organic acids were monitored. HPLC was used to confirm the organic acids produced in the cultures as metabolites. L. plantarum YK-9 produced both lactic acid and acetic acid, which were responsible for the pH decrease during growth. Initial pH 7.0 of the cultures decreased to 3.6 at the incubation after 72 hours, and concentrations of lactic acid and acetic acid increased to approximately 588.7 mM and 255.5 mM, respectively. The antibacterial activities against food-poisoning causing bacteria were examined with 20-fold concentrated culture supernatants from L. plantarum YK-9, and the antibacterial effects were clearly observed against all the bacteria tested in this work.

Influence of Temperature on the Bacterial Community in Substrate and Extracellular Enzyme Activity of Auricularia cornea

  • Zhang, Xiaoping;Zhang, Bo;Miao, Renyun;Zhou, Jie;Ye, Lei;Jia, Dinghong;Peng, Weihong;Yan, Lijuan;Zhang, Xiaoping;Tan, Wei;Li, Xiaolin
    • Mycobiology
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    • v.46 no.3
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    • pp.224-235
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    • 2018
  • Temperature is an important environmental factor that can greatly influence the cultivation of Auricularia cornea. In this study, lignin peroxidase, laccase, manganese peroxidase, and cellulose in A. cornea fruiting bodies were tested under five different temperatures ($20^{\circ}C$, $25^{\circ}C$, $30^{\circ}C$, $35^{\circ}C$, and $40^{\circ}C$) in three different culture periods (10 days, 20 days and 30 days). In addition, the V4 region of bacterial 16S rRNA genes in the substrate of A. cornea cultivated for 30 days at different temperatures were sequenced using next-generation sequencing technology to explore the structure and diversity of bacterial communities in the substrate. Temperature and culture days had a significant effect on the activities of the four enzymes, and changes in activity were not synchronized with changes in temperature and culture days. Overall, we obtained 487,694 sequences from 15 samples and assigned them to 16 bacterial phyla. Bacterial community composition and structure in the substrate changed when the temperature was above $35^{\circ}C$. The relative abundances of some bacteria were significantly affected by temperature. A total of 35 genera at five temperatures in the substrate were correlated, and 41 functional pathways were predicted in the study. Bacterial genes associated with the membrane transport pathway had the highest average abundance (16.16%), and this increased at $35^{\circ}C$ and $40^{\circ}C$. Generally, different temperatures had impacts on the physiological activity of A. cornea and the bacterial community in the substrate; therefore, the data presented herein should facilitate cultivation of A. cornea.

Thrombotic thrombocytopenic purpura with decreased level of ADAMTS-13 activity and increased level of ADAMTS-13 inhibitor in an adolescent (청소년기에 발생한 ADAMTS-13 활성도 저하와 항체 양성을 보인 혈전저혈소판혈증자색반병 1례)

  • Yang, Eun Mi;Han, Dong Kyun;Baek, Hee Jo;Shin, Myung Geun;Kim, Young Ok;Kook, Hoon;Hwang, Tae Ju
    • Clinical and Experimental Pediatrics
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    • v.53 no.3
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    • pp.428-431
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    • 2010
  • Thrombotic thrombocytopenic purpura (TTP) is a thrombotic microangiopathy characterized by endothelial cell damage, resulting in microangiopathic hemolytic anemia, thrombocytopenia, and various degrees of neurological and renal impairment caused by microvascular thrombi. It is rare in children and frequently follows a fatal course. TTP is divided into 2 types: one is inherited and associated with ADAMTS-13 gene mutations and the other is acquired and associated with anti-ADAMTS-13 autoantibodies. The measurement of ADAMTS-13 activity in plasma, identification of ADAMTS-13 circulating inhibitor, anti-ADAMTS-13 IgG, and ADAMTS-13 gene sequencing are crucial to the diagnosis of TTP. Plasma exchanges are the first-line treatment for acquired TTP, combined with steroids and immunosuppressive drugs. Here, we describe the case of an adolescent patient with TTP, confirmed by decreased level of ADAMTS-13 activity and an increased level of ADAMTS-13 inhibitor, who was successfully treated by plasma exchanges.

Identification of a New Agar-hydrolyzing Bacterium Vibrio sp. S4 from the Seawater of Jeju Island and the Biochemical Characterization of Thermostable Agarose (제주도 연안 해양에서 분리한 한천분해 미생물 Vibrio sp. S4의 동정 및 내열성 agarase의 생화학적 특성)

  • Lee, Chang-Ro;Chi, Won-Jae;Bae, Chang-Hwan;Hong, Soon-Kwang
    • Microbiology and Biotechnology Letters
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    • v.43 no.4
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    • pp.314-321
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    • 2015
  • Agar-hydrolyzing bacteria were isolated from the coastal sea water of Jeju Island. One isolate, designated as S4, was selected for further study. The S4 cells were Gram-negative and rod-shaped with smooth beige surfaces and single polar flagellum. Cells were grown at $15-42^{\circ}C$, 0.5-5% (w/v) NaCl, between pH 6.0 and 9.0, and in media containing 0.5-5% (w/v) NaCl. The G+C content was 49.93 mol%. The major fatty acids (>15%) were $C_{18:1}{\omega}7c$, $C_{16:0}$ and Summed feature 3 (comprising $C_{16:1}{\omega}7c/iso-C_{15:0}$ 2-OH). Based on 16S rRNA sequencing and biochemical and chemotaxonomic characteristics, the strain was designated as Vibrio sp. S4. In liquid culture supplemented with 0.1% agar the cell density and agarase activity reached a maximum level in 72 h, while agarase activity in the culture without agar was negligible, implying agarose expression is induced by agar. The optimum pH and temperature for the extracellular crude agarase of S4 were 7.0 and $45^{\circ}C$, respectively. However, it also exhibited 98.6% and 87.6% at $40^{\circ}C$ and $50^{\circ}C$, respectively, of the maximum activity seen at $45^{\circ}C$. The crude agarase hydrolyzed agarose into (neo)agarotetraose and (neo)agarohexaose.

Probiotic Properties of Lactic Acid Bacteria isolated from Feces and Kimchi (베트남인 분변 및 김치로부터 분리된 유산균의 프로바이오틱스 기능성 연구)

  • Shin, Hyun Su;Yoo, Sung Ho;Jang, Jin Ah;Won, Ji Young;Kim, Cheol Hyun
    • Journal of Dairy Science and Biotechnology
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    • v.35 no.4
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    • pp.255-261
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    • 2017
  • The purpose of this study was to investigate the probiotic properties and antioxidant capacity of lactic acid bacteria isolated from Vietnamese feces and the Korean traditional food kimchi. Six isolated strains were identified as Lactobacillus sp. by 16S rRNA sequencing. All strains showed good resistance to low pH (1.5, 2.0, and 3.0) and 0.3% oxgall bile acids. Culture filtrates from the six strains showed various antioxidant effects, including DPPH, ABTS, reducing power, and metal chelating ($Fe^{2+}$) activities. Two of the six Lactobacillus strains showed potential probiotic activity. Heat resistance and adhesion assays were conducted by mixing the selected strains, Lactobacillus acidophilus V4, Lactobacillus plantarum V7, and Lactobacillus paracasei DK121 isolated from kimchi. The results showed that the heat resistance of these strains was similar to that of a commercial strain, L. plantarum LP. In addition, a mucin attachment assay using the mixture of selected strains (V4, V7, and DK121) showed high binding activity to the mucous layer. In conclusion, a mixture of V4, V7, and DK121 shows promising probiotic activity and may be useful for the development of health-related products.

Isolation and Degradation Activity of a TBTCl (Tributyltin Chloride) Resistant Bacteriain Gwangyang Bay (광양만에서 TBTCl (Tributyltin Chloride) 내성세균의 분리 및 분해활성)

  • Jeong, Seong-Yun;Son, Hong-Joo;Jeoung, Nam-Ho
    • Korean Journal of Environmental Agriculture
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    • v.30 no.4
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    • pp.424-431
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    • 2011
  • BACKGROUND: Tributyltin chloride is among the most toxic compounds known for aquatic ecosystems. Microorganisms are responsible for removal of TBTCl. Nevertheless, only a limited number of marine bacteria were investigated for biodegradation of TBTCl in Korea. METHODS AND RESULTS: The number of TBTCl resistant bacteria ranged from $2.5{\times}10^3$ to $3.8{\times}10^3$ cfu/mL in the seawater, and ranged from $3.2{\times}10^5$ to $9.1{\times}10^5$ cfu/g in the surface sediment, respectively. The morphological, physiological, and biochemical characteristics of TBTCl resistant bacteria were investigated by API 20NE and other tests. The most abundant species of TBTCl resistant bacteria were Vibrio spp. (19.2%), Bacillus spp. (16.2%), Aeromonas spp. (15.2%), and Pseudomonas spp. (13.1%), etc. Eleven TBTCl resistant isolates also had a resistance to heavy metals (Cd, Cu, Hg, and Zn). Among them, isolate T7 showing the strong TBTCl-resistance was selected. This isolate was identified as the genus Pantoea by 16S rRNA gene sequencing and designated as Pantoea sp. T7. In addition, this bacterium was cultivated up to the growth of 50.7% after 60 hrs at TBTCl concentration of $500{\mu}M$. TBTCl-degrading activity of Pantoea sp. T7 was measured by GC-FPD analysis. As a result of biological TBTCl-degradation at TBTCl concentration of $100{\mu}M$, TBTCl-removal efficiency of Pantoeasp. T7 was 62.7% after 40 hrs. CONCLUSION(S): These results suggest that Pantoea sp. T7 is potentially useful for the bioremediation of TBT contamination.