• Applied Biological Chemistry
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• v.13 no.1
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• pp.51-57
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• 1970

On the basis of the specific interrelationship between the species or variety of leguminous crops and the species or strain of nodule bacteria, Rhizobia, the rhizobial species and strain must be effectively chosen for the successful inoculation. The present paper describes on some results of the isolation and taxonomic study on the native rhizobial strains isolated from the nodules of five species of leguminous crops such as numerous varieties of soy bean, lespedeza, birdfoot trefoil, ladino and red clovers. The isolated strains of soy bean nodule bacterium, Rhizobium japonicum were grouped through the inoculation test on variety Changdanbaikmock into the effective, noneffective and toxic strain for the nodule formation. In the study of the effect of some inorganic and organic nitrogenous compounds on the growth of Rhizobium japonicum strain Ac 20, a promotive response was showed by asparagine, and glutamine, but hydroxylamine, nitrite, hydrazine and azide was inhibitory at the concentration of $10^{-2}M/l$ in mannitol-yeast extract basal medium. In the physiological characteristics each strain showed somewhat different activities of the indole-3-actic acid formation and hydrogenase and discussed with these characters in relation to nodule forming ability.

• Journal of Microbiology and Biotechnology
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• v.33 no.2
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• pp.188-194
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• 2023

Microbacterium elymi KUDC0405^T was isolated from the rhizosphere of Elymus tsukushiensis from the Dokdo Islands. The KUDC0405^T strain was Gram-stain-positive, non-spore forming, non-motile, and facultatively anaerobic bacteria. Strain KUDC0405^T was a rod-shaped bacterium with size dimensions of 0.3-0.4 × 0.7-0.8 ㎛. Based on 16S rRNA gene sequences, KUDC0405^T was most closely related to Microbacterium bovistercoris NEAU-LLE^T (97.8%) and Microbacterium pseudoresistens CC-5209^T (97.6%). The dDDH (digital DNA-DNA hybridization) values between KUDC0405^T and M. bovistercoris NEAU-LLE^T and M. pseudoresistens CC-5209^T were below 17.3% and 17.5%, respectively. The ANI (average nucleotide identity) values among strains KUDC0405^T, M. bovistercoris NEAU-LLE^T, and M. pseudoresistens CC-5209^T were 86.6% and 80.7%, respectively. The AAI (average amino acid identity) values were 64.66% and 64.97%, respectively, between KUDC0405^T and its closest related type strains. The genome contained 3,596 CDCs, three rRNAs, 46 tRNAs, and three non-coding RNAs (ncRNAs). The genomic DNA GC content was 70.4%. The polar lipids included diphosphatydilglycerol, glycolipid, phosphatydilglycerol, and unknown phospholipid, and the major fatty acids were anteiso-C_17:0 and iso-C_16:0. Strain KUDC0405^T contained MK-12 as the major menaquinone. Based on genotypic, phylogenetic, and phenotypic properties, strain KUDC0405^T should be considered a novel species within the genus Microbacterium, for which we propose the name M. elymi sp. nov., and the type strain as KUDC0405^T (=KCTC 49411^T, =CGMCC1.18472^T).

• Journal of Microbiology and Biotechnology
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• v.26 no.2
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• pp.337-346
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• 2016

Three combinations of molecular chaperones from Escherichia coli (i.e., DnaK-DnaJ-GrpE-GroEL-GroES, GroEL-GroES, and DnaK-DnaJ-GrpE) were overproduced in E. coli BL21, and their in vitro stabilizing effects on a nitrile hydratase (NHase) were assessed. The optimal gene combination, E. coli groEL-groES (ecgroEL-ES), was introduced into Rhodococcus ruber TH3. A novel engineered strain, R. ruber TH3G was constructed with the native NHase gene on its chromosome and the heterologous ecgroEL-ES genes in a shuttle plasmid. In R. ruber TH3G, NHase activity was enhanced 37.3% compared with the control, TH3. The in vivo stabilizing effect of ecGroEL-ES on the NHase was assessed using both acrylamide immersion and heat shock experiments. The inactivation behavior of the in vivo NHase after immersion in a solution of dynamically increased concentrations of acrylamide was particularly evident. When the acrylamide concentration was increased to 500 g/l (50%), the remaining NHase activity in TH3G was 38%, but in TH3, activity was reduced to 10%. Reactivation of the in vivo NHases after varying degrees of inactivation was further assessed. The activity of the reactivated NHase was more than 2-fold greater in TH3G than in TH3. The hydration synthesis of acrylamide catalyzed by the in vivo NHase was performed with continuous acrylonitrile feeding. The final concentration of acrylamide was 640 g/l when catalyzed by TH3G, compared with 490 g/l acrylamide by TH3. This study is the first to show that the chaperones ecGroEL-ES work well in Rhodococcus and simultaneously possess protein-folding assistance functions and the ability to stabilize and reactivate the native NHases.

• Korean Journal of Microbiology
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• v.36 no.1
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• pp.26-32
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• 2000

Catechol 2,3-dioxygenase was purified from recombinant strain E. coli CNU312 carrying the tomB gene which was cloned from toluene-degrading Burkholderia cepacia G4. The purification of this enzyme was performed by acetone precipitation, Sephadex G-75 chromatography, electrophoresis and electro-elution. The molecular weight of native enzyme was about 140.4 kDa and its subunit was estimated to be 35 kDa by SDS-PAGE. It means that this enzyme consists of four identical subunits. This enzyme was specifically active to catechol, and$K_(m)$ value and $V_(max)$value of this enzyme were 372.6 $\mu$M and 39.27 U/mg. This enzyme was weakly active to 3-methylcatechol, 4-methylcatechol, and 4-chlorocatechol, but rarely active to 2,3-DHBP. The optimal pH and temperature of the enzyme were pH 8.0 and $40^{\circ}C$. The enzyme was inhibited by $Co^(2+)$, $Mn^(2+)$, $Zn^(2+)$, $Fe^(2+)$, $Fe^(3+)$, and $Cu^(2+)$ ions, and was inactivated by adding the reagents such as N-bromosuccinimide, and $\rho$-diazobenzene sulfonic acid. The activity of catechol 2,3-dioxygenase was not stabilized by 10% concentration of organic solvents such as acetone, ethanol, isopropyl alcohol, ethyl acetate, and acetic acid, and by reducing agents such as 2-mercaptoethanol, dithiothreitol, and ascorbic acid. The enzyme was inactivated by the oxidizing agent $H_(2)$$O_(2)$, and by chelators such as EDTA, and ο-phenanthroline.

• Journal of Animal Science and Technology
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• v.45 no.6
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• pp.1019-1030
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• 2003

The study was conducted to isolate and identify highly fibrolytic anaerobic fungi from the guts of a Hanwoo steer and a Korean native goat, and then investigate the characterization of cellulolytic activity of an anaerobic fungus. Twenty-one anaerobic fungal colonies were isolated in the study, in which 16 colonies were isolated from the rumen contents of the Hanwoo steer and 5 colonies from the duodenal fluids of the Korean native goat. Four anaerobic fungi were selected based on higher cellulolytic enzyme activities to identify under a optical microscope. NLRI-M003 and -T004 belong to Neocallimastix genus and NLRI-M014 belongs to Piromyces genus based on the morphology of their thallus, sporangia, rhizoid and the number of flagella. NLRI-M001 appeared to be an unknown strain of anaerobic fungi due to its different morphology from existing types of anaerobic fungi, though the morpholgoy is similar to Orpinomyces sp. Supplementation of 2% anaerobic fungal culture(NLRI-M003) in rumen-mixed microorganisms increased in vitro DM degradability of rice straw and filter paper up to 4 and 11%, respectively, compared with non-supplementation(control). CMCase and xylanase activities in in vitro culture were also higher in 2% fungal supplementation than controls in both rice straw and filter paper substrates.

• Korean Journal of Agricultural Science
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• v.44 no.4
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• pp.531-540
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• 2017

A study was conducted to compare growth performance of six female commercial Korean native chicken (KNC) crossbreeds from hatching to twelve weeks of age. Three hundred and twelve, 1-day-old female commercial KNC were used within 1 paternal line and 6 maternal lines. The chickens were allocated to 24 battery cages to give 4 replicates per strain with 13 chickens per cage. The chickens were reared under continuous lighting (24 h) and water was available at all times. Ad-libitum feeding was practiced throughout the experimental period. Among the six different strains, 2A had the greatest bodyweight (BW) at 42 days after hatching (p < 0.05). No BW difference between six crossbreed strains (p > 0.05) was found thereafter. Crossbreed 1A had the higher average daily gain (ADG) than crossbreed 2A and 3A chickens (p < 0.05), whereas crossbreed 4A, 5A, and 6A had similar ADGs to that of crossbreed 1A (p > 0.05) at 84 days after hatching. Furthermore, crossbreed 4A had a great average daily feed intake (ADFI) from hatching to 84 days (p < 0.05). Nonetheless, there was no difference in the feed conversion ratio (FCR) and uniformity between six crossbreed strains for the experimental period (p > 0.05). Despite that 1A, 4A, and 6A had the higher viability (p < 0.05) than crossbreed 2A and 5A, they had a similar viability than crossbreed 3A (p > 0.05). With this in mind, crossbreed 2A had greater BW, ADG, and FCR than other chicken crossbreeds from hatching to 84 days, although they had a lower viability than others.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.21 no.11
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• pp.102-108
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• 2020

This study was conducted to investigate the genetic diversity and genetic taxonomic relationships between Korean native black goat (KNBG) populations and crossbred goats. The 45,658 common single nucleotide polymorphisms present in the KNBG strain and crossbred goat were used for the analysis. The expected and observed heterozygosity (which can be indicators of genetic diversity) were in the order of crossbred, Gyeongsang National University, Jangsu, then the Tongyeong strains. The variance component represents the degree of genetic diversity between groups. The highest variance (19.98 %) was between the Dangjin and Gyeongsang National University strains. The lowest variance (8.87 %) was between the Jangsu and Tongyeong strains. In addition, the genetic distance between the populations showed that Jangsu and Tongyeong formed one branch (they were very similar genetically). The Dangjin and the Gyeongsang National University strains appeared to form a second branch. Furthermore, the crossbred formed one branch with the Dangjin and the Gyeongsang National University strains. Therefore, the results of this study can be used as basic data to reduce unnecessary inbreeding and genetic resource flow between the KNBG populations. The basic data indicates the uniqueness of the genetic resources of the domestic lineage. These findings provide a basis for differentiating KNBG and Crossbred goats to use to improve the desirable characteristics of this species.

• Journal of Life Science
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• v.21 no.11
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• pp.1619-1624
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• 2011

Endophytic fungal strains were isolated from the roots of six species plants in the Dokdo islands. Native plant samples, such as Artemisia japonica, Chenopodium album and Solanum nigrum were isolated from Dongdo, and those such as Cyrtomium falcatum, Dianthus longicalyx and Tetragonia tetragonoides were isolated from Seodo. In total, thirty two fungal strains were isolated from these native plants. To identify the fungal strains, polymerase chain reaction (PCR) amplification of internal transcribed spacer (ITS: containing ITS1, 5.8s and ITS2 region) regions was done with universal primers ITS1 and ITS4. Endophytic fungi of four species were isolated from A. japonica, eight species from C. album, three species from S. nigrum, three species from C. falcatum, three species from D. longicalyx and eleven species from T. tetragonoides. Culture filtrates (CF) of isolated endophytic fungi were used to treatwaito-c rice seedlings to test plant growth-promoting activity. As a result of bioassay, Ca-5-2-2 strain isolated from C. album expressed highest plant growth-promotion activity. Of all the endophytic fungi isolated, Penicillium sp., Fusarium sp. and Aspergillus sp. were the most abundantly distributed fungal strains in the six plants used in this study.

• Journal of Life Science
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• v.12 no.1
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• pp.77-86
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• 2002

Chitin, a $\beta$-1,4 polymer of N-acetyl-D-glucosamine, is one of the most abundant organic compounds in nature. Chitinase (EC 3.2.1.14) is an enzyme that degrades chitin to chito-oligosaccharides, diacetyl rhitobiose and N-acetyl-D-glucosamine. An extracellular chitinase-producing bacterial strain was isolated from soil and named to as Bacillus subtilis JK-56. Optimum culture condition of B. subtilis JK-56 for the production of chitinase was 1% chitin, 0.5% polypepton, 0.1% KCl, 0.05% MnS $O_4$.4$H_2O$, 37$^{\circ}C$, initial pH 7.0 and 40 hour culture time. When B. subtilis JK-56 was grown in the optimum medium, one major active band and two minor active bands were detected by native-PAGE and active staining of the gel. Among them, the major band was purified from the culture supernatant by 70% ammonium sulfate precipitation and native-PAGE with BIO-RAD Model 491 Prep-Cell and named as Chi-56A. Its molecular weight was estimated to be 53kDa monomer and the isoelectric point (pI) was pH 4.3. The pH and temperature for the optimum activity of Chi-56A were pH 6.0 and $65^{\circ}C$, respectively. Chi-56A was stable up to $65^{\circ}C$ and in alkaline region. Its $K_{m}$ value for colloidal chitin was 17.33g/L. HPLC analysis of the reaction products confirmed that Chi-56A was an exo type chitinase.e.

• Journal of Microbiology and Biotechnology
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• v.5 no.2
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• pp.68-73
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• 1995

Pseudomonas sp. P20 was shown to be capable of degrading biphenyl and 4-chlorobiphenyl (4CB) to produce the corresponding benzoic acids wnich were not further degraded. But the potential of the strain for biodegradation of 4CB was shown to be excellent. The pcbA, B, C and D genes responsible for the aromatic ring-cleavage of biphenyl and 4CB degradation were cloned from the chromosomal DNA of the strain. In this study, the pebC and D genes specifying degradation of 2, 3-dihydroxybiphenyl (2, 3-DHBP) produced from biphenyl by the pebAB-encoded enzymes were cloned by using pBluescript SK(＋) as a vector. From the pCK102 (9.3 kb) containing pebC and D genes, pCK1022 inserted with a EcoRI-HindIII DNA fragment (4.1 kb) carrying pebC and D and a pCK1092 inserted with EcoRI-XbaI fragment (1.95 kb) carrying pebC were constructed. The expression of pcbC and D' in E. coli CK102 and pebC in E. coli CK1092 was examined by gas chromatography and UV-vis spectrophotometry. 2.3-dihydroxybiphenyl was readily degraded to produce meta-cleavage product (MCP) by E. coli CK102 after incubation for 10 min, and then only benzoic acid(BA) was detected in the 24-h old culture. The MCP was detected in E. coli CK1022 containing pebC and 0 genes (by the resting cells assay) for up to 3 h after incubation and then diminished completely in 8 h, whereas the MCP accumulated in the E. coli CK1092 culture even after 6 h of incubation. The 2, 3-DHBP dioxygenases (product of pebC gene) produced by E. coli CK1, CK102, CK1023, and CK1092 strains were measured by native PAGE analysis to be about 250 kDa in molecular weight, which were about same as those of Pseudomonas sp. DJ-12, P. pseudoa1caligenes KF707, and P. putida OU83.