• Journal of Microbiology and Biotechnology
• /
• v.10 no.2
• /
• pp.275-280
• /
• 2000

The inulin fructotransferse (depolymerizing) (IFTase, EC 2.4.1.93) gene of Arthrobacter sp. A-6 was cloned and expressed in Escherichia coli and Bacillus subtilis. The IFTase gene consisted of an ORF of 1.311 nucleotides encoding a polypeptide of 436 amino acids containing a signal peptide of 31 amino acids in the N-terminus. The molecular mass of the IFTase based on the nucleotide sequence was calculated to be 46.116 Da. The recombinant E. coli $DH5{\alpha}$ cells expressing the Arthrobacter sp. A-6 IFTase gene produced most of the IFTase intracelularly. In contrast, the recombinant B. subtilis DB 104 carrying the IFTas gene on a B. subtilis-E. Coli expression vector secreted the IFTase into the culture fluid efficiently.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.45 no.1
• /
• pp.35-42
• /
• 2012

We investigated the effect of dietary supplementation with probiotics on growth performance, feed utilization, innate immunity and disease resistance against Streptococcus iniae in juvenile olive flounder Paralichthys olivaceus. Four experimental diets were made by supplementation with three Bacillus sp. probiotics (B. subtilis, B. pumilus and B. licheniformis) into a basal diet (control) at 0.5% each. Three-hundred fish (average BW, 20 g) were randomly distributed (25 per tank) into twelve 150 L capacity polyvinyl circular tanks with three replicates per dietary treatment. Triplicate groups of fish were fed the experimental diets to apparent satiation (twice per day, 09:00 and 17:00 h) for nine weeks. After the feeding trial, a challenge test was directly conducted against S. iniae. The results indicated that dietary supplementation with the probiotic, Bacillus sp, seemed to improve growth performance of olive flounder. Hematological parameters and innate immunity as determined by nitroblue tetrazolium, myeloperoxidase or superoxide dismutase activities were not significantly affected by the probiotic supplementation. The result from the challenge test suggested that dietary supplementation with probiotics Bacillus sp. can improve the disease resistance of olive flounder against S. iniae.

• Korean Journal of Microbiology
• /
• v.42 no.1
• /
• pp.67-72
• /
• 2006

The nucleotide sequence of the cloned cellulolytic xylanase gene (bglBC2) from B. circulans ATCC21367 was determined. bglBC2 consists of an 1,224 bp open reading frame (ORF) coding for a polypeptide of 407 amino acids with a deduced molecular weight of 45 kDa. The Shine-Dalgarno (SD) sequence (5'-AAAGGAG-3') was found 9 bp upstream of the initiation codon, ATG. A promoter region corresponding closely to the B. subtilis consensus sequence (-35: TTGACA,-10: TATAAT) was detected, the putative -35 and -10 sequences of which were TTTACA and TATACT, respectively. The deduced amino acid sequence of the cellulolytic xylanase showed 97% homology with that of the alkaline $endo-\beta-1,4-glucanase$ from B. circulans KSM-N257, 75% homology with that of the $endo-\beta-1,3-1,4-glucanase$ from B. circulans WL-12, and 45% homology with that of the $endo-\beta-1,4-glucanase$ (cellulase) from Bacillus sp. KSM-330. The bglBC2 sequence was deposited in Gen-Bank under the accession number AY269256.

• Korean Journal of Environmental Biology
• /
• v.28 no.2
• /
• pp.95-100
• /
• 2010

Burkholderia cepacia PBSA-7, Bacillus licheniformis PBSA-8 and Burkholderia sp. PBSA-9 previously collected from Korea soil (Joo and Kim, 2009) were analyzed for the presence of genes encoding proteins operative in the degradation of poly(butylene succinate-co-butylene adipate; PBSA). Polymerase chain reaction analyses revealed a 1.5 kb fragment of the lipase gene (lip A) in B. cepacia PBSA-7 and Burkholderia sp. PBSA-9, while B. licheniformis PBSA-8 harbored the same gene fragment at 600 bp. The three strains possessed "Gly-X1-Ser-X2-Gly" and "Ala-X1-Ser-X2-Gly" lipase sequence regions. Burkholderia sp. PBSA-7 lip A displayed 36~40% homology with the family 1-1 lipases and 82~92% homology with the family 1-5. Burkholderia sp. PBSA-8 lip A was 64~65% homologous with the subfamily 1-4 lipases, but displayed no homology with the subfamily 1-5 lipases. Burkholderia sp. PBSA-9 lip A displayed 35~37% homology with the family I1 lipases and 83~94% homology with the family I2 lipases, similar to Burkholderia sp. PBSA-7.

• Journal of Microbiology and Biotechnology
• /
• v.10 no.2
• /
• pp.244-250
• /
• 2000

An ${\alpha}-glucosidase$ gene (aglA) from thermophilic Bacillus sp. DG0303 was cloned, sequenced, and expressed in Escherichia coli. The aglA was localized to the 2.1-kb PvuI-XmnI region within the 5.9-kb DNA insert of the gybrid plasmid pAG1. The gene consisted of an open reading frame of 1,686 bp with an unusual GTG initiation codon and TGA termination codon. The amino acid sequence deduced from the nucleotide sequence predicted a protein of 562 amino acid residues with a M, of 66,551 dalton. A comparative amino acid sequence analysis revealed that DG0303 ${\alpha}-glucosidase$ is related to bacillary oligo-1, 6-glucosidases. The Bacillus sp. DG0303 ${\alpha}-glucosidase$ showed a high sequence identity (36-59%) to the B. flavocaldarius, B. cereus, and B. thermoglucosidasius oligo-1, 6-glucosidases. The number of prolines in theses four ${\alpha}-glucosidases. was observed to increase with increasing thermostability of these enzymes. The cloned ${\alpha}-glucosidase was purified from E. coli $DH5{\alpha}$ bearing pAG1 and characterized. The recombinant enzyme was identical with the native enzyme in its optimum pH and in its molecular mass, estimated by sodium dodecy1 sulfate-polyacrylamide gel electrophoresis. The temperature optimum of the cloned ${\alpha}-glucosidase$ was lower than that of the native enzyme.

• Journal of Microbiology and Biotechnology
• /
• v.8 no.1
• /
• pp.28-33
• /
• 1998

A gene coding for ${\beta}$-xylosidase from thermophilic xylanolytic Bacillus sp. KK-1 was cloned into Escherichia coli using plasmid pBR322. Recombinant plasmid DNAs were isloated from E. coli clones which were capable of hydrolyzing 4-methylumbelliferyl-${\beta}$-D xylopyranoside. Restriction analysis showed the DNAs to share a common insert DNA. Xylo-oligosaccharides, including xylotriose, xylotetraose, xylopentaose, and xylobiose were hydrolyzed to form xylose as an end product by cell-free extracts of the E. coli clones, confirming that the cloned gene from strain KK-1 is ${\beta}$-xylosidase gene. The ${\beta}$-xylosidase gene of strain KK-1 designated as xylB was completely sequenced. The xylB gene consisted of an open reading frame of 1,602 nucleotides encoding a polypeptide of 533 amino acid residues, and a TGA stop codon. The 3' flanking region contained one stem-loop structure which may be involved in transcriptional termination. The deduced amino acid sequence of the KK-1 ${\beta}$-xylosidase was highly homologous to the ${\beta}$-xylosidases of Bacillus subtilis and Bacillus pumilus, but it showed no similarity to a thermostable ${\beta}$-xylosidase from Bacillus stearothermophilus.

• Korean Journal of Food Science and Technology
• /
• v.38 no.1
• /
• pp.82-87
• /
• 2006

Twenty one strains strongly producing protease were isolated from Korean traditional Cheonggukjang. Eight strains selected by sensory evaluation on Cheonggukjang prepared with isolated strains were identified with based on biochemical properties a and 16S rDNA sequencing. Identified strains were Bacillus subtilis MB4, and Bacillus amyloliquefaciens A1, A2, B1, MC1, SB2, SC1, and SD1. Protease activities, important strain selection factor, were higher in Cheongjukjang prepared with B. subtilis MB4 (179.6 Unit) and B. amyloliquefaciens SB2 (201.9 Unit) than commercial traditional Cheonggukjang (97.9 Unit). Sensory evaluation revealed Cheonggukjang prepared with B. subtilis MB4 had flavor very similar to commercial traditional Cheonggukjang. Cheonggukjang prepared with B. suhtilis MB4 (0.0006 Pa s) and commercial traditional Cheonggukjang (0.0002 Pa s) revealed lower viscosities than those of Cheonggukjang prepared with B. amyloliquefaciens SB2, MC1, B1, A1, SD1, A2, and SC1 (0.006 to 0.008 Pa s at 1001/s. Results show Cheonggukjang could be prepared using single strain of B. subtilis MB4, maintaining high protease activity and very similar sensory and viscosity qualities with those of commercial traditional Cheonggukjang.

• International Journal of Industrial Entomology and Biomaterials
• /
• v.37 no.1
• /
• pp.9-14
• /
• 2018

Bacterial species, Bacillus amyloliquefaciens and Lactobacillus species (L. sp.5-1), are known to inhibit the growth of pathogenic bacteria and fungi. Metarhizium anisopliae is a pathogenic fungal species which causes fatal damage to P. brevitarsis populations. Therefore, we investigated the inhibitory effect of B. amyloliquefaciens and L. sp. 5-1 on M. anisopliae induced fatality rates in P. brevitarsis. Samples of M. anisopliae-infected sawdust were treated with strain B. amyloliquefaciens KACC10116, strain L. sp. 5-1 KACC19351, and a combination of the two. P. brevitarsis were fed treated sawdust samples, and their subsequent fatality rate was monitored. The fatality rate fell below 1.5% after 10 days and decreased by approximately 40% after 15 days. On average, the fatality rate decreased by 20%, compared to the control. The difference in the decrease in fatality rate between B. amyloliquefaciens treatment and L. sp. 5-1 treatment was not significant. Results indicate that both strains exhibit high anti-fungal activity, which may be useful in environmental purification efforts. These strains may be used for effective prevention of fungal infection in P. brevitarsis.

• Journal of Life Science
• /
• v.20 no.2
• /
• pp.269-274
• /
• 2010

An antifungal antibiotic-producing strain, BCNU 2003, was isolated from forest soil in Korea. The morphological and physiological characters, and 16S rRNA sequences analysis of strain BCNU 2003 identified this strain as Bacillus genus. The Bacillus sp. BCNU 2003 showed strong antifungal activities against Aspergillus niger, Trichophyton mentagrophytes and Trichophyton rubrum with inhibition ranging from 62.05 to 63.49% by using dual culture technique. Bacillus sp. BCNU 2003 produced a maximum level of antifungal substances under aerobic incubation at 28oC and pH 6.5-7.2 for 6 days in LB broth. Ethyl acetate extract of the cultured broth showed strong antifungal activity and a broad antifungal spectrum against various pathogenic fungi. The minimum inhibitory concentration (MIC) values for its active extracts ranged between 0.0625 mg/ml and 1 mg/ml. In addition, Bacillus sp. BCNU 2003 was determined to have the ability to produce enzymes such as amylase, protease, gelatinase and catalase.

• The Plant Pathology Journal
• /
• v.39 no.5
• /
• pp.430-448
• /
• 2023

Recently, strategies for controlling Fusarium oxysporum f. sp. lycopersici (Fol), the causal agent of Fusarium wilt of tomato, focus on using effective biocontrol agents. In this study, an analysis of the biocontrol and plant growth promoting (PGP) attributes of 11 isolates of loamy soil Bacillus spp. has been conducted. Among them, the isolates B.PNR1 and B.PNR2 inhibited the mycelial growth of Fol by inducing abnormal fungal cell wall structures and cell wall collapse. Moreover, broad-spectrum activity against four other plant pathogenic fungi, F. oxysporum f. sp. cubense race 1 (Foc), Sclerotium rolfsii, Colletotrichum musae, and C. gloeosporioides were noted for these isolates. These two Bacillus isolates produced indole acetic acid, phosphate solubilization enzymes, and amylolytic and cellulolytic enzymes. In the pot experiment, the culture filtrate from B.PNR1 showed greater inhibition of the fungal pathogens and significantly promoted the growth of tomato plants more than those of the other treatments. Isolate B.PNR1, the best biocontrol and PGP, was identified as Bacillus stercoris by its 16S rRNA gene sequence and whole genome sequencing analysis (WGS). The WGS, through genome mining, confirmed that the B.PNR1 genome contained genes/gene cluster of a nonribosomal peptide synthetase/polyketide synthase, such as fengycin, surfactin, bacillaene, subtilosin A, bacilysin, and bacillibactin, which are involved in antagonistic and PGP activities. Therefore, our finding demonstrates the effectiveness of B. stercoris strain B.PNR1 as an antagonist and for plant growth promotion, highlighting the use of this microorganism as a biocontrol agent against the Fusarium wilt pathogen and PGP abilities in tomatoes.