• Journal of Microbiology and Biotechnology
• /
• v.4 no.2
• /
• pp.102-107
• /
• 1994

The $\beta$-1,4-endoglucanase gene from Actinomyces sp. 40 was cloned into Escherichia coli DH5$\alpha$ with pUC19. Chromosomal DNA from Actinomyces sp. 40 was cleaved with the restriction enzyme Sau3AI and ligated into pUC19 for the transformation of Escherichia coli DH5$\alpha$. Positive clones of $\beta$-1,4-endoglucanase gene were detected as the clear zones on a medium supplemented with carboxymethylcellulose (CMC). This transformant possessed a single plasmid, designated pDS1, which contained the vector DNA and a 3.5 kilobase (kb) Sau3AI insertion fragment encoding endoglucanase. The size of the cloned fragment was reduced to 2.0 kb. The endoglucanase activity produced by the E. coli DH5$\alpha$ (pDS6) was higher than that of Actinomyces sp. 40 strain. The optimum pH and temperature of the cloned enzyme were pH 4.0$\sim$5.0 and 55$^{\circ}C$, respectively. The cloned enzyme was stable at 55$^{\circ}C$ or below and in buffer ranging from pH 4.0 to 7.0. The enzyme degraded CMC but did not degrade xylan, cellobiose, and methyl-umbelliferylcellobiopyranoside (MUC).

• Asian-Australasian Journal of Animal Sciences
• /
• v.7 no.3
• /
• pp.373-382
• /
• 1994

A bacterial strain No. 40, which produced extracellular endoglucanase, was isolated from the rumen of Korean native goals and identified to be a genus of Actinomyces sp. The optimum conditions for endoglucanase production in PY-CMC medium were initial pH of 7.0 and 4 days of cultivation at $39^{\circ}C$. When localization of endoglucanase activity of Actinomyces sp. was determined, 68% of the enzyme activity was found in the extracellular fraction, 11% of the activity was detected in the periplasmic space and the remaining activity was in the intracellular and cell-bound fractions. The maximal endoglucanase activity was observed at pH 5.0 and it was most s table at pH 5.0. The optimum temperature of this enzyme activity was $55^{\circ}C$, but enzyme activity was gradually lost at temperature above $60^{\circ}C$. The crude enzyme was activated by addition of 10 mM cysteine and 10 mM DTT. But it was inhibited by addition of 10 mM $Cu^{{+}{+}}$ and $Fe^{{+}{+}}$. This crude enzyme could digest carboxymethylcellulose (CMC), and degrade xylan, avicel, pNPG, and pNPC to a less extent.

• Journal of Korean society of Dental Hygiene
• /
• v.13 no.5
• /
• pp.889-900
• /
• 2013

Objectives : Molecular biology techniques were employed to assess diversity of bacterial in children's dental caries. Methods : DNA of germs was extracted and the diversity of the 16S rRNA clones was analyzed by amplified rDNA restriction analysis and sequencing. The experimental samples were pit and fissure caries (PC), deep dentinal caries (DC), smooth surface caries (SC), and supragingival plaque (PQ) from 50 children of age less than 12 years old. The control group was healthy teeth supragingival plaque (HT). Thirty clones from each 16S rRNA clone library of 5 samples were randomly selected, thus a total of 150 clones were analyzed. Results : Amplified rDNA restriction analysis uncovered 18, 20, 11, 17, and 22 phylotypes from healthy teeth, pit and fissure caries, deep dentinal caries, smooth surface caries, and supragingival plaque, respectively. Sequencing analysis found the dominance of Actinomycs naeslundii and Fusobacterium nucleatum in the healthy teeth; Leptotrichia sp. in the pit and fissure caries; Actinomyces sp., Streptococcus mutans, and Rahnella aquatilis in the deep dentinal caries; Streptococcus mutans and Actinomyces sp. in the smooth surface caries; Enterobacter hormaechei and Streptococcus sanguinis in the supragingival plaque. Conclusions : Clonal analysis identified 6 phyla, 20 genera, and 51 species.

• Journal of the Korea Organic Resources Recycling Association
• /
• v.22 no.4
• /
• pp.54-61
• /
• 2014

This study was conducted to investigate the effect of Actinomyces sp. and Bacillus sp., United States granular microorganisms and Japan granular microorganisms on turfgrass growth and thatch decomposing of creeping bentgrass in golf course by measuring turf color index, chlorophyll index, thatch content of soil, root length, turf density and chemical properties and thatch content of soil. Fertilizer treatment was designed as follows; control(CF; compound fertilizer), microorganism medium(M; CF+M), microorganism medium and livestock manure fertilizer(M-L; CF+M+LMF), microorganism medium, livestock manure fertilizer and amino acid liquid fertilizer(M-L-A; MM+LMF+ALF), United States granular microorganisms(USGM; CF+USGM), Japan granular microorganisms(CF+JGM). Soil properties investigated after experiment was scarcely affected by applied fertilizers in root zone of creeping bentgrass. The turf color index and chlorophyll index of M, M-L, M-L-A, USGM, JGM treatment were higher than those of CF. The turfgrass root in M-L treatment was longer than others. The thatch content of soil in M treatment was longer than others. The thatch content of M was decreased than that of CF by 6.8%. These was suggested that application of M induced the development of quality and growth of creeping bentgrass by assisting turfgrass growth and thatch decomposing.

• Microbiology and Biotechnology Letters
• /
• v.46 no.1
• /
• pp.59-67
• /
• 2018

A xylanolytic and cellulolytic anaerobic bacterium strain CtC72 was isolated from cattle rumen liquor. Phylogenetic analysis of 16S rRNA gene sequences revealed that strain CtC72 shared only 97.78% homology with its nearest phylogenetic affiliate Actinomyces ruminicola, showing its novelty. The strain could grow on medium containing xylan, carboxymethyl cellulose and avicel producing $CO_2$, acetate, and ethanol as major fermentation products. The whole genome analysis of the strain CtC72 exhibited a broad range of carbohydrate-active enzymes required for the breakdown and utilization of lignocellulosic biomass. Genes related to the production of ethanol and stress tolerance were also detected. Further there were several unique genes in CtC72 for chitin degradation, pectin utilization, sugar utilization, and stress response in comparison with Actinomyces ruminicola. The results show that the strain CtC72, a putative novel bacterium can be used for lignocellulosic biomass based biotechnological applications.

• Journal of Technologic Dentistry
• /
• v.21 no.1
• /
• pp.139-144
• /
• 1999

The optimum culture conditions for an antibiotics from Actinomyces sp. were investigated. The optimum composition of medium for antibiotics production was 1% glucose, 1% soybean meal, 0.5% NaCl, 0.1% $CaCO_2$, and the optimum initial pH was 7.0. And the antibiotics showed highest activity when the strain isolated from soil was aerobically cultivated at $28^{\circ}C$ for 72hours under the optimum conditions. A production of the antibiotics from Actinomyces sp. begins at the 36th hours and then reached the maximum at the stationary phase developed at the 72th hours under the optimum conditions.

• Asian-Australasian Journal of Animal Sciences
• /
• v.29 no.1
• /
• pp.126-133
• /
• 2016

A gene from Actinomyces sp. Korean native goat (KNG) 40 that encodes an endo-${\beta}$-1,4-glucanase, EG1, was cloned and expressed in Escherichia coli (E. coli) $DH5{\alpha}$. Recombinant plasmid DNA from a positive clone with a 3.2 kb insert hydrolyzing carboxyl methyl-cellulose (CMC) was designated as pDS3. The entire nucleotide sequence was determined, and an open-reading frame (ORF) was deduced. The ORF encodes a polypeptide of 684 amino acids. The recombinant EG1 produced in E. coli $DH5{\alpha}$ harboring pDS3 was purified in one step using affinity chromatography on crystalline cellulose and characterized. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis/zymogram analysis of the purified enzyme revealed two protein bands of 57.1 and 54.1 kDa. The amino terminal sequences of these two bands matched those of the deduced ones, starting from residue 166 and 208, respectively. Putative signal sequences, a Shine.Dalgarno-type ribosomal binding site, and promoter sequences related to the consensus sequences were deduced. EG1 has a typical tripartite structure of cellulase, a catalytic domain, a serine-rich linker region, and a cellulose-binding domain. The optimal temperature for the activity of the purified enzyme was $55^{\circ}C$, but it retained over 90% of maximum activity in a broad temperature range ($40^{\circ}C$ to $60^{\circ}C$). The optimal pH for the enzyme activity was 6.0. Kinetic parameters, $K_m$ and $V_{max}$ of rEG1 were 0.39% CMC and 143 U/mg, respectively.

• Asian-Australasian Journal of Animal Sciences
• /
• v.18 no.1
• /
• pp.61-65
• /
• 2005

The objective of this work was to investigate the nutritional requirements for the growth of Actinomyces sp. 9RCC5 isolated from the rumen of a native goat in Korea. The growth of strain 9RCC5 on the basal medium or the medium minus certain ingredients from the basal medium demonstrated that strain 9RCC5 showed absolute requirement of vitamin B complex mixture, while hemin and volatile fatty acids (VFA) were stimulatory to growth to some extent. The 9RCC5 strain grew well with casein hydrolysate as the sole added nitrogen source. However, neither a complex of 18 amino acids nor ammonium sulfate effectively replaced casein hydrolysate. Vitamins such as riboflavin and pantothenate were essential for growth, while thiamin and biotin were stimulatory. With regard to VFA, the growth was stimulated by acetic acid but inhibited by valeric acid. Relatively large quantities of $Na^+$, $K^+$ and $Ca^{2+}$ were absolutely required for growth. Supplementation of clarified rumen fluid to the basal medium in a range of 0-10% (vol/vol) resulted in an increased rate of growth as well as an increased extent of growth.

• 한국생물공학회:학술대회논문집
• /
• 2005.10a
• /
• pp.364-367
• /
• 2005

This study investigated the production of antioxidant from Actinomyces culture supernatant. For the research of the natural marine antioxidant, several bacteria were isolated from the coast of Je-ju in Korea. An actinomycetes strains, S-1 was identified to a genus level 16S ribosomal DNA sequence and fatty acid analysis. From these results and other characteristics described in the Bergey's Manual, this strain was identificated as a Nocardiopsis dassonvillei. Strain S-1 showed high activity of 1,1-diphenyl-2-prcrylhydrazyl radical scavenging. The hydroxyl radical scavenging ability of Nocardiopsis sp. S-1 supernatant was 53%. Nutritional and cultural conditions for the production of antioxidant by this organism under shake-flask conditions have optimized. Similary initial medium pH 7.6, incubation temperature of $25^{\cicr}C$, sodium chloride concentration 2.5 and incubation time of 8 day were found to be optimal.

• Korean Journal of Environmental Agriculture
• /
• v.20 no.1
• /
• pp.63-66
• /
• 2001

To investgate growth effect of tomato by Bacillus sp. WRD-1 isolated from soil, the Bacillus sp. WRD-1 cultures were treated into tomato cultivated soil with different dilutions (1:100, 1:300, and 1:500) and autoclaved Bacillus cultures as control. Growth and yeild of tomato enhanced in treatments of the Bacillus cultures compared to control. The populations of native bacteria and actinomyces were increased twice in field treated with Bacillus sp. WRD-1 cultures, but the number of mold was decreased. Since the Bacillus sp. WRD-1 promoted growth of tomato and affected population dynamics of microorganism in field, this strain is prominent candidate as a microbial biocide to improve soil potential.