• Korean Journal of Microbiology
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• v.54 no.2
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• pp.126-135
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• 2018

The characteristics of enzyme and gene for mannanase B had been reported from Cellulosimicrobium sp. YB-43 producing some kind of mannanase. A gene coding for the enzyme, named mannanase C (ManC), was expected to be located downstream of the manB gene. The manC gene was cloned by polymerase chain reaction and sequenced completely. From this nucleotide sequence, ManC was identified to consist of 448 amino residues and contain a carbohydrate binding domain CBM2 besides a catalytic domain, which was homologous to mannanase belonging to the glycosyl hydrolase family 5. The catalytic domain of ManC showed the highest amino acid sequence similarity of 55% with the mannanases from Streptomyces sp. SirexAA-E (55.8%; 4FK9_A) and S. thermoluteus (57.6%; BAM62868). The His-tagged ManC (HtManC) lacking N-terminal signal peptide with hexahistidine at C-terminus was produced and purified from cell extract of recombinant Escherichia coli. The purified HtManC showed maximal activity at $65^{\circ}C$ and pH 7.5, with no significant change in its activity at pH range from 7.5 to 10. HtManC showed more active on konjac and locust bean gum (LBG) than guar gum and ivory nut mannan (ivory nut). Vmax and Km values of the HtManC for LBG were 68 U/mg and 0.45 mg/ml on the optimal condition, respectively. Mannobiose and mannotriose were observed on TLC as major products resulting from the HtManC hydrolysis of mannooligosacharides. In addition, mannobiose and mannose were commonly detected as the hydrolyzed products of LBG, konjac and ivory nut.

• Journal of Microbiology and Biotechnology
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• v.17 no.5
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• pp.721-727
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• 2007

Stability-enhanced mutants, H44, 11-94, 5A2-84, and F8, of L-threonine aldolase(L-TA) from Streptomyces coelicolor A3(2)(SCO1085) were isolated by an error-prone PCR followed by a high-throughput screening. Each of these mutant, had a single amino acid substitution: H177Y in the H44 mutant, A169T in the 11-94 mutant, D104N in the 5A2-84 mutant and F18I in the F8 mutant. The residual L-TA activity of the wild-type L-TA after a heat treatment for 20 min at $60^{\circ}C$ was only 10.6%. However, those in the stability-enhanced mutants were 85.7% for the H44 mutant, 58.6% for the F8 mutant, 62.1% for the 5A2-84 mutant, and 67.6% for the 11-94 mutant. Although the half-life of the wild-type L-TA at $63^{\circ}C$ was 1.3 min, those of the mutant L-TAs were longer: 14.6 min for the H44 mutant, 3.7 min for the 11-94 mutant, 5.8 min for the 5A2-84 mutant, and 5.0 min for the F8 mutant. The specific activity did not change in most of the mutants, but it was decreased by 45% in the case of mutant F8. When the aldol condensation of glycine and 3,4-dihydroxybenzaldehyde was studied by using whole cells of Escherichia coli containing the wild-type L-TA gene, L-threo-3,4-dihydroxyphenylserine(L-threo-DOPS) was successfully synthesized with a yield of 2.0 mg/ml after 20 repeated batch reactions for 100 h. However, the L-threo-DOPS synthesizing activity of the enzyme decreased with increased cycles of the batch reactions. Compared with the wild-type L-TA, H44 L-TA kept its L-threo-DOPS synthesizing activity almost constant during the 20 repeated batch reactions for 100 h, yielding 4.0 mg/ml of L-threo-DOPS. This result showed that H44 L-TA is more effective than the wild-type L-TA for the mass production of L-threo-DOPS.

• Korean Journal of Soil Science and Fertilizer
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• v.31 no.4
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• pp.420-426
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• 1998

This investigation was undertaken to clarify the effects of application of chemical fertilizers (Compound fertilizer and commercial compost) and organic matters (straw and clover) on changes of the composition of soil actinomycetes flora. Actinomycetes were isolated from chemical fertilizer and organic materials treated soils, and then grown on HV agar plate. The isolated strains were classified by Bergey's manual based on the morphological characteristics and color of substrate mycelium of actinomycetes. The number of actinomycetes increased 4 times in control, 36 times in clover, 20 times in straw, 5 times in chemical fertilizer and 4 times in commercial compost treated soil after 14th day of incubation. This result suggests that the application of straw or clover is more effective to increase the number of actinomycetes than compound fertilizer or commercial compost. It also showed that the application of chemical fertilizer or organic materials considerably changed the composition of soil actinomycetes flora. The proportion of streptomyces strains to the isolated total actinomycetes was 62% in control, 60% in clover, 68% in straw, 67% in chemical fertilizer and 64% in commercial compost treated soils at the 14th day.

• Korean Journal of Microbiology
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• v.54 no.3
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• pp.272-279
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• 2018

This study was carried out to evaluate antimicrobial activity of isolated bacteria from various soils against two strains of Propionibacterium acnes causing acne vulgaris. Among several hundreds of bacterial strains, Paenibacillus elgii DS381, Paenibacillus elgii DS1515, Burkholderia gladioli DS518, and Streptomyces lienomycini DS620 showed high antimicrobial activities against the strains of P. acnes. All isolated bacteria showed 15.5 to 34.3 mm inhibition zone diameter in an agar well diffusion test, and especially DS620 showed the highest inhibition zone diameters (28.3~34.3 mm). Antibacterial substances were expected as lipopeptide (pelgipeptin and paenipeptin) from strains DS381 and DS1515, protease from DS518, and anthracycline antibiotic (daunomycinone) from DS620, and all these showed very low minimum inhibitory concentration [DS381 and DS1515 (0.078 mg/ml), DS518 (0.312 mg/ml), DS620 (0.000078 mg/ml)] against P. acnes. These antibacterial substances could completely kill P. acnes within 24 h in a time-kill assay. These results suggest that antibacterial substances produced by these bacteria may be utilized as useful antimicrobial agent against P. acnes and treatment medicine for acne vulgaris.

• Korean Journal of Plant Tissue Culture
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• v.25 no.4
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• pp.283-288
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• 1998

The bialaphos is a potent inhibitor of glutamine synthease in higher plants and is used as a non-selective herbicide. We have used the bialaphos resistant gene(Bar) encoding for an acetyltransferase isolated from Streptomyces hygroscopicus SF1293. Callus derived from mature seeds of rice(Oryza sativa L. cv. Dong Jin) were co-cultivated with Agrobacterium tumefaciens EHA101 carring a plasmid pGPTV-HB containing genes for hygromycin resistance (HygR) and Bar. Transgenic plants showing in vitro resistance to 50 mg/L hygromycin and 10 mg/L bialaphos were obtained by using a two-step selection/regeneration procedure. Transformation efficiency of rice was about 30% which was as high as reported in other dicotyledons. Progenies ($\textrm{T}_{1}$ generation) derived from primary transformant of 17 lines were segregated with a 3 resistant : 1 sensitive ratio in medium containing hygromycin and bialaphos. Stable integration of Bar gene into chromosomal DNA was proven by Southern blot analysis of genomic DNA isolated from $\textrm{T}_{2}$ progenies. Transgenic plants ($\textrm{T}_{3}$) grown in the field were resistant to bialaphos (Basta) at a dosage lethal to wild type plants.

• Korean Journal of Organic Agriculture
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• v.22 no.4
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• pp.719-730
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• 2014

This study was conducted to select proper soil amendments in order to reduce the occurrence of potato scab and maintaining soil health by applications of dolomitic lime, sulfur, potassium sulfate, shell meal fertilizer, silicate fertilizer, lime nitrogen and ammonium sulfate fertilizer in different pH levels of volcanic ash soil with continuous cultivation of potato. In potassium sulfate-applied plot with a low soil pH, the incidence rate and disease severity of scab were lowest at 84.4% and 28.4%, respectively. Those were lowest among the treatments. Value of potato scab control was 12.3% and marketable yield of potato was highest at 93.2%. In lime nitrogen-applied plot (60 kg/10a), the incidence rate was low at 38.3%, and control value was 23.8% and marketable yield of potato was high at 66.3%. In relatively higher pH soils, the incidence rate of scab was lowest at 38.3% in the lime nitrogen-applied plot (60 kg/10a). Value of potato scab control was 23.8%, which was four times higher than that in sulfur-applied plot. Marketable yield of potato was highest at 66.3% in the lime nitrogen-applied plot. In the lime nitrogen plot infected with potato scab pathogen such as S. acidiscabies and S. scabiei were remarkably lower than other soil amendments at 2.5, 5, and 10 g/L concentrations of lime nitrogen using Glucose Yeast Malt (GYM) medium. In conclusion, this study suggests that potassium sulfate application in low pH soil (less than pH 5) and lime nitrogen application in relatively higher pH soil (more than pH 6) before potato seeding might be helpful to reduce the occurrence of potato scab.

• Journal of Life Science
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• v.12 no.1
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• pp.113-119
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• 2002

The cloning and expression of Phosphotyrosine Protein Phosphatase into E. coli provides important tools of understanding of its functions and signal transduction mechanisms. The abundant soluble protein of the Phosphotyrosine Protein Phosphatase A (PtpA) and the active site mutant PtpA(C9S) were produced using the expression vector pET26 in E. coli and pIJ6021 with the thiostrepton in S. lividans. The enzyme activity of both proteins extracted by Ni-NTA column had same results from the expression vector pET26 and pIJ6021. The enzyme activity of phosphatase was found in the protein of PtpA, but not in that of C9S. The western blot detected by penta His-tag antibody resulted in the inducer, thiostrepton was not a good trigger to induce a large amount of PtpA protein. The overexpression of both proteins had no significantly different effect on the A factor cascade related to the secondary metabolite and mycelium formation between PtpA and C9S. However, overproduction of PtpA protein using pIJ6021 in S. lividans brought about a dramatic decrease in the amount of phosphotyrosine proteins (p200, p90, and p65), but no significantly phenotypic variation in S. lividans. This indicates that PtpA has an important proteome role in signal transduction mechanism of producing massive amount of phosphotyrosine protein in Streptomyces sp.

• Current Research on Agriculture and Life Sciences
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• v.14
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• pp.101-110
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• 1996

Using response surface methodology(RSM), the various conditions(agitation speed, air flow, glucose concentration) in jar fermentor culture were investigated to find the optimum conditions for maximum enzyme production. Central-composite-design was used to control the variable constant in the experiment. The glucose isomerase production of Steptomyces chibaensis J-59 was mostly affected by the air flow rate and glucose concentration. The estimated optimum conditions were as follows: 1% birchwood xylan, 1.5% CSL, 0.1% $MgSO_4{\cdot}7H_2O$, 0.012% $CoCl_2{\cdot}6H_2O$, pH 7.0; air flow, 2.2vvm; agitation speed, 587rpm; glucose concentration, 0.586%. Experimental values(7.43GIU/ml) for the enzyme production obtained from the given optimum conditions had a almost resemblane to response values(7.67GIU/ml) predicted by the RSM. The jar fermentor culture by the RSM produced xylose isomerase about 2.7 times as much as the baffled flask culture.

• Journal of the Korean Applied Science and Technology
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• v.36 no.3
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• pp.748-757
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• 2019

The objective of the present study was to evaluate effects of probiotic mixture (PM) including Bacillus subtillus, Streptomyces galilaeus and Sphingobacteriaceae on egg production, egg quality, and eliminating odor in laying hens. A total of 240 Hy-Line Brown laying hens (50 wks of age) were divided into four treatment groups (60 replicates of birds, each) in completely randomized design. Groups were assigned to four experimental diets: CON (a basal diet of no PM additive as control), basal diet supplemented with either CP3 (0.3% commercial probiotics), PM3 (0.3% PM), or PM5 (0.5% PM 0.5). Data of egg production and egg quality were obtained during 6 weeks of experimental period. Egg production, Haugh unit, shell thickness, breaking strength, yolk color, and albumin high were significantly increased in CP3 and PM3 compared to those in CON and CP3. Ammonia levels and numbers of E. coli, total aerobic bacteria, Coliform, and Salmonella in feces were significantly increased in order of CON>CP3>PM3>PM5. The present study indicates that addition of 0.3% probiotic mixture including B. subtillus, S. galilaeus and Sphingobacteriaceae to diets may improve egg production, egg quality, and eliminate fecal ammonia of laying hens.

• Journal of Microbiology and Biotechnology
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• v.18 no.4
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• pp.658-662
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• 2008

Effects of pH shock on the secretion system of S. coelicolor A3(2) have been investigated at a transcriptional level by using DNA microarrays. Actinorhodin secretion was observed to be highly enhanced when an acidic-pH shock was applied to surface grown cultures of S. coelicolor A3(2). In this culture, a gene of actVA-orf1 encoding a putative efflux pump or transporter protein for actinorhodin was strongly upregulated. A major number of efflux pumps for other metabolites and a major number of secretion proteins for protein secretion were also observed to be upregulated with pH shock. The secretion of actinorhodin was observed to be remarkably enhanced in liquid culture as well.