• 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.

• Korean Journal of Poultry Science
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• v.44 no.3
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• pp.189-198
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• 2017

This study was conducted to develop a new synthetic breed of Korean native chicken. The combining ability and reciprocal effects for production traits were estimated on 1,157 hens from a $5{\times}5$ diallel cross-mating design using grand parent stock (GPS) lines of Korean native chicken. Body weight, viability, age at first egg laying, egg weight, and hen-day egg production were measured and analyzed. The results showed that the general combining ability (GCA) of the survival rate during laying periods was -9.6 to 11.1, with the highest value obtained in the W strain. Additionally, the GCA of the body weight at 12 weeks was -209.7 to 162.2, with the highest value obtained in the F strain. The GCA for age at fist egg laying was estimated to be -2.8 to 3.7, while the GCA of egg weight was -0.91 to 0.96, and the GCA of hen-day egg production was -4.9 to 6.0. In the estimation of specific combining ability, the YW combination showed the highest survival rate, FW showed the highest body weight at 12 weeks, and GW showed the highest hen-day egg production. The reciprocal effects were significantly different among crosses for almost all productivity traits. In identical breeding combinations, differences in ability were observed when the maternal or paternal breeds were switched. The mean value based on combining ability was higher in WY, WF, and GW combinations for survival rate; GF, HG, and HF combinations for body weight at 12 weeks; and GW, YW, and FW combinations for hen-day egg production. It is concluded that the GF and HF combinations, which have excellent growth performance and moderate survival rate, are the most desirable paternal parent stock (PS) strains, and the GW and FW combinations, which have great laying performance and moderate body weight, are the most desirable maternal PS strains.

• Korean Journal of Poultry Science
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• v.44 no.2
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• pp.123-134
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• 2017

This study was conducted to establish new synthetic lines of Korean Native Chicken. We performed $5{\times}5$ diallel crossings with GPS lines of Korean Native Chicken for the selection of parent stock. The production traits including viability, body weight, age at first egg laying, egg weight, hen-day egg production, and hen-housed egg production were measured and analyzed for 25 crosses with 1,157 hens. The heterosis effects of these traits were also estimated. The results showed that the average survival rate during laying periods was 67.7% in the pure lines and 77.1% in the crosses. The 25 cross combinations were shown to be distinctly divided into three groups according to body weight; nine crosses in the high-weight group, 12 crosses in the medium-weight group and four crosses in the light-weight group. The average body weight at 12 weeks of age was $1,873.8{\pm}43.0g$ in the high group, $1,595.4{\pm}56.6g$ in the medium group and $1,152.7{\pm}24.7g$ in the light group, and $1,560.2{\pm}339.3g$ in the pure lines and $1,640.9{\pm}213.7g$ in the crosses. In terms of egg production performance, the age at first egg laying was $139.7{\pm}4.9$ days in the pure lines and $135.8{\pm}5.1$ days in the crosses. The average egg weight was $52.3{\pm}1.6g$ in the pure line and $53.0{\pm}1.6g$ in the crosses. The average hen-housed egg production from 20 to 40 weeks was $58.3{\pm}12.9$ eggs in the pure line and $69.2{\pm}10.4$ eggs in the crosses. The average heterosis effect was 16.9% for survival rate, 5.4% for body weight at 12 weeks, -2.7% for age at first egg laying, 1.3% for egg weight, and 14.3% for hen-day egg production. Generally, the heterosis effect for viability and egg production traits was higher than that for growth trait. On the basis of these results, it is expected that the selection of the HF combination, which had excellent growth performance, is the most desirable as the paternal strain, and the selection of GW and FW combinations, which had excellent laying performance, are preferable as the maternal strain.

• 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.

• Journal of Microbiology and Biotechnology
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• v.24 no.2
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• pp.254-263
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• 2014

Enteromorpha polysaccharides (EP) extracted from green algae have displayed a wide variety of biological activities. However, their high molecular weight leads to a high viscosity and low solubility, and therefore, greatly restrains their application. To solve this problem, bacteria from the surface of Enteromorpha were screened, and an Alteromonas macleodii strain B7 was found to be able to decrease the molecular weight of EP in culture media. Proteins harvested from the supernatant of the A. macleodii B7 culture were subjected to native gel electrophoresis, and a band corresponding to the Enteromorpha polysaccharide lyase (EPL) was detected by activity staining. The enzyme identity was subsequently confirmed by MALDI-TOF/TOF mass spectrometry as the putative ${\alpha}$-amylase reported in A. macleodii ATCC 27126. The amylase gene (amySTU) from A. macleodii B7 was cloned into Escherichia coli, resulting in high-level expression of the recombinant enzyme with EP-degrading activity. AmySTU was found to be cold-adapted; however, its optimal enzyme activity was detected at $40^{\circ}C$. The ${\alpha}$-amylase was highly stable over a broad pH range (5.5-10) with the optimal pH at 7.5-8.0. The highest enzyme activity was detected when NaCl concentration was 2%, which dropped by 50% when the NaCl concentration was increased to 16%, showing an excellent nature of halotolerance. Furthermore, the amylase activity was not significantly affected by tested surfactants or the presence of some organic solvents. Therefore, the A. macleodii strain B7 and its ${\alpha}$-amylase can be useful in lowering EP molecular weight and in starch processing.