• Journal of Life Science
• /
• v.15 no.5 s.72
• /
• pp.734-738
• /
• 2005

A gene coding for xylanase from alkali-tolerant Bacillus alcalophilus AX2000 was cloned into Escherichia coli $DH5\alpha$ using pUC19. Among 2,000 transformants, one transformant showed clear zone on the detection agar plate containing oat-spells xylan. Its recombinant plasmid, named pXTY99, was found to carry 7.0 kb insert DNA fragment. When the nucleotide sequence of the cloned xylanase gene (xynT) was determined, xynT gene was found to consist of 1,020 base-pair open reading frame coding for a poly-peptide of 340 amino acids with a deduced molecular weight of 40 kDa. The coding sequence was preceded by a putative ribosome binding site, and the transcription initiation signals. The deduced amino acid sequence of xylanase is similar to those of the xylanases from Bacillus sp. Nl37 and B. stearothermophilus 21 with $61\%$ and $59\%$ identical residues, respectively.

• Korean Journal of Soil Science and Fertilizer
• /
• v.34 no.6
• /
• pp.387-393
• /
• 2001

To use as a fundamental data for the sustainable agriculture, which is nowadays a major trend to keep the productivity and conserve the environment, 487 paddy soil samples were collected from 21 regions of the Gyeongnam Province and analyzed the chemical characteristics and microbial population of the soil. The microbial population densities were bacteria $298{\times}10^5$($4{\sim}3000{\times}10^5$ range), fungi $63{\times}10^3$($2{\sim}441{\times}10^3$ range), actinomycetes $19{\times}10^5$($0.2{\sim}1250{\times}10^5$ range), Bacillus sp. $53{\times}10^4$($4{\sim}890{\times}10^4$ range) and Pseudomonas sp. $198{\times}10^4CFU\;g^{-1}$($4{\sim}1724{\times}10^4CFU\;g^{-1}$ range), respectively. The microbial populations of the soil were in general higher in southern area than in the northern area of the Gyeongnam Province. The average ratio of bacteria/fungi population was 473. As soil clay content increased, the populations of aerobic bacteria, actinomycetes and Pseudomonas sp. were remarkably decreased. The ratio of aerobic bacteria and fungi was 1554 in sandy loam and clay loam 1144, while Bacillus sp./fungi ratio was 11 in clay loam and 10 in loam. On the topographical differences, aerobic bacteria and Bacillus sp./fungi ratio were the higher in coastal plains than any other areas. The microbial population densities from different soil types were generally lower in ill-drained paddy field than those of other paddy field. The content of $P_2O_5$, K, Ca, $NO_3-N$ and EC in soil were positively correlated to the population densities of aerobic bacteria, actinomycetes, fungi, Bacillus sp. and Pseudomonas sp.. The soil organic matter and Mg content were also positively correlated to the population densities of aerobic bacteria, actinomycetes, fungi and Bacillus sp.

• Asian-Australasian Journal of Animal Sciences
• /
• v.27 no.4
• /
• pp.495-502
• /
• 2014

The effect of Bacillus subtilis natto on hindgut fermentation and microbiota of early lactation Holstein dairy cows was investigated in this study. Thirty-six Holstein dairy cows in early lactation were randomly allocated to three groups: no B. subtilis natto as the control group, B. subtilis natto with $0.5{\times}10^{11}cfu$ as DMF1 group and B. subtilis natto with $1.0{\times}10^{11}cfu$ as DMF2 group. After 14 days of adaptation period, the formal experiment was started and lasted for 63 days. Fecal samples were collected directly from the rectum of each animal on the morning at the end of eighth week and placed into sterile plastic bags. The pH, $NH_3$-N and VFA concentration were determined and fecal bacteria DNA was extracted and analyzed by DGGE. The results showed that the addition of B. subtilus natto at either treatment level resulted in a decrease in fecal $NH_3$-N concentration but had no effect on fecal pH and VFA. The DGGE profile revealed that B. subtilis natto affected the population of fecal bacteria. The diversity index of Shannon-Wiener in DFM1 decreased significantly compared to the control. Fecal Alistipes sp., Clostridium sp., Roseospira sp., beta proteobacterium were decreased and Bifidobacterium was increased after supplementing with B. subtilis natto. This study demonstrated that B. subtilis natto had a tendency to change fecal microbiota balance.

• The Korean Journal of Mycology
• /
• v.44 no.3
• /
• pp.176-183
• /
• 2016

Bacillus velezensis CC112 inhibited the mycelial growth of several plant pathogens, including Rhizoctonia solani, causing damping-off on ginseng. The control efficacies of B. velezensis CC112 against R. solani by seed dipping in LB and BSM broth diluted 10 times, soil dipping, and soil drenching with LB broth diluted 10 times were 65.8%, 67.1%, and 64.2%, respectively. Treatment of soil drenching with the 100 times diluted prototype of B. velezensis CC112 against R. solani and Pythium sp. by soil revealed control efficacies of 77.3% and 65.7%, respectively. These results indicate that B. velezensis CC112 is a prospective biofungicide for the biological control of ginseng damping off.

• The Journal of the Korean Society for Microbiology
• /
• v.35 no.1
• /
• pp.31-40
• /
• 2000

Bacillus anthracis, the etiological agent of anthrax has been classified into the Bacillus subgroup I with B. cereus, B. mycoides and B. thuringiensis based on morphological and DNA similarity. DNA studies have further indicated that these species have very AT-rich genomes and high homology, indeed it has been proposed that these four sub-species be recognized as members of the one species. Several methods have been developed to obtain good differentiation between these species. However, none of these methods provides the means for an absolutely correct differntiation. The analysis of fatty acid methyl esters (FAMEs) was employed as a quick, simple and reliable method for the identification of 21 B. anthracis strains and closley related strains. The most significant differences were found between B. anthracis and B. anthracis closely related strains in FAMEs profiles. All tested strains of B. anthracis had a branched fatty acid C17:1 Anteiso A, whereas the fraction of unsaturated fatty acid Iso C17:1 w10c was found in B. anthracis closely related strains. By UPGMA clustering analysis of FAMEs profiles, all of the tested strains were classified into two clusters defined at Euclidian distance value of 24.5. The tested strains of B. anthracis were clustered together including Bacillus sp. Kyungjoo 3. However, the isolates of B. anthracis closely related spp. Rho, S10A, 11R1, CAU9910, CAU9911, CAU9912 and CAU9913 were clustered with the other group. On the basis of these results, isolates of B. anthracis Bongchon, Kyungjoo 1, 2 and Bacillus sp. Kyungjoo 3 were reclassified as a B. anthracis. It is concluded that FAMEs analysis provides a sensitive and reliable method for the identification of B. anthracis from closely related taxa.

• Microbiology and Biotechnology Letters
• /
• v.16 no.5
• /
• pp.343-347
• /
• 1988

The promoters of alkali-tolerant Bacillus sp. had been cloned in the promoter probe vector pPL703 and recombinant plasmid p-12 had been constructed. As a result of subcloning, two different promoters were found to exist in the cloned 2.9 kb promoter fragment and two recombinant plasmids p-l2B1 and p-l2B2, each harboring different promoter, were constructed. The promoter activity, which was expressed in the CAT specific activity, of p-l2B1 was 7 times higher than that of p-l2B2. The promoter activity as a function of growth revealed that both promoters of p-l2B1 and p-l2B2 were expressed after the late logarithmic growth phase and repressed in the presence of 1.0％ (w/v) glucose.

• Journal of Korean Ophthalmic Optics Society
• /
• v.18 no.1
• /
• pp.67-73
• /
• 2013

Purpose: Recently, bacterial contamination of equipment and accessories required for vision correction has become a main causal factor in ophthalmic diseases. Thus, We investigated on both the actual condition of bacterial contamination from glasses of vision correction. Methods: Investigation of microorganisms was carried out with a group of 145 glasses wearers, composed of 36 elementary school students, 37 middle school students, 38 high school students, 10 college students, and 32 aged men. Results: Seventeen species of bacteria are detected from glasses of vision correction: B. cereus, B. licheniformis, Bacillus sp., CNS, Enterococcus sp., Escherichia coli, Proteus sp., Pseudomonas sp., Serretia sp., Streptococcus sp., Staphylococcus epidermidis, Staphylococcus aureus, Streptococcus hemolyticus,, Acinetobacter sp., Enterobacter cloacae, GNR, and Pseudomonas aeruginosa. Among 17 species of bacteria, there are some potential causative agents for keratitis, corneal ulcer, Acute dacryocystitis, Orbital cellulitis, Periphlebitis retinae, Marginal blepharitis, and Acute conjunctivitis. Enterobacter cloacae, Pseudomonas aeruginosa and Staphylococcus epidermidis cause keratitis. Pseudomonas sp., and Staphylococcus aureus cause corneal ulcer. Staphylococcus aureus causes acute dacryocystitis, orbital cellulitis, periphlebitis retinae, marginal belpharitis. Streptococcus hemolyticus causes acute conjunctivitis. Conclusions: In summation, it is verified that hazardous, opportunistic and infectious microorganisms exist in glasses for vision correction. Ophthalmic diseases are predicted. Therefore, supplementary research on the development of a cleaning solution to cleanse the infection and of an effective method to remove microorganisms is required.

• Microbiology and Biotechnology Letters
• /
• v.39 no.4
• /
• pp.374-381
• /
• 2011

The beneficial effects of Eisenia fetida on soil properties have been attributed to their interaction with soil microorganisms. The bacterial diversity of the intestinal tract of E. fetida was investigated by culture-dependent and culture-independent methods including denaturing gradient gel electrophoresis (DGGE) and pyrosequencing analyses. In a pure culture, Lysinibacillus fusiformis (51%), Bacillus cereus (30%), Enterobacter aerogenes (21%), and L. sphaericus (15%) were identified as the dominant microorganisms. In the DGGE analyses, B. cereus (15.1%), Enterobacter sp. (13.6%), an uncultured bacterium (13.1%), and B. stearothermophilus (7.8%) were identified as the dominant microorganisms. In the pyrosequencing analyses, Microbacterium soli (26%), B. cereus (10%), M. esteraromaticum (6%), and Frigoribacterium sp. (6%) were identified as the dominant microorganisms. The other strains identified were Aeromonas sp., Pseudomonas sp., Borrelia sp., Cellulosimicrobium sp., Klebsiella sp., and Leifsonia sp. The results illustrate that culture independent methods are better able to detect unculturable microorganisms and a wider range of species, as opposed to isolation by culture dependent methods.

• Microbiology and Biotechnology Letters
• /
• v.17 no.5
• /
• pp.436-439
• /
• 1989

The chromosomal DNA fragments of thermophilic alkalophilic Bacillus sp, K-17, a potent xylanhydrolyzing bacterium, were ligated to a vector plasmid pBR322 and transformed into Escherichia coli HB101. The plasmid pAX278, isolated from a transformant forming yellow color on the LB agar plate containing 1 mM p-nitrophenyl- $\beta$-xylopyranoside, was found to enable the transformants to produce p-xylosidase. The 5.0 kilobase insert of pAX278 had single sites for EcoRI, PstI, XbaI, and PvuII, and 2 sites for BglII. Biotinylated pAX218 was hybridized to 0.9 kb as well as 5.0 kb fragment from Bacillus sp. K-17 DNA on nitrocellulose filter. pGX718 was constructed by inserting the 5.0 kb HindIII fragment of pGX278 at the HindIII site of pGR71, E. coli and B. subtilis shuttle vector. The enzymatic properties of $\beta$-xylosidase from E. coli HB101 carrying recombinant plasmid were the same those of $\beta$-xylosidase from Bacillus sp. K-17.

• Microbiology and Biotechnology Letters
• /
• v.31 no.3
• /
• pp.216-223
• /
• 2003

A 1.3-kb of chitosanase gene (choA) encoding 45-kDa polypeptide was cloned, expressed, and characterized from a newly isolated Bacillus cereus H-1. The chitosanase protein (ChoA) of B. cereus H-l was purified to homogeneity by ammonium sulfate precipitation and CM-sephadex column chromatography. Optimum pH was around 7, and stable pH range in the incubation at 50 C was 4-11. Optimum temperature was around 50 C, and enzyme activity was relatively stable below 45 C. ChoA showed the activities toward carboxymethyl cellulose (CMC) in addition to soluble or glycol chitosan. Based on MALDI-TOF MS analysis of purified ChoA, the entire amino acid sequence of ChoA was interpreted by database searching of previously known Bacillus chitosanases. A 1.6 kb of PCR product of corresponding chitosanase gene was obtained and its DNA sequence was determined. The deduced amino acid of choA revealed that ChoA have a 98% homology with those of Bacillus sp. No.7-M strain and Bacillus sp. KCTC0377BP. The recombinant ChoA protein was expressed in E. coli DH5$\alpha$. Deduced amino acid comparison of choA with other chitosanases suggested that it belongs to family 8 microbial endo-chitosanase with chitosanase-cellulase activity.