• Fisheries and Aquatic Sciences
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• v.6 no.1
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• pp.1-6
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• 2003

A chitinolytic enzyme-producing bacterium was isolated from sea water on the coast of Busan. The bacterium was identified as Aeromonas sp. based on its morphological, cultural and biochemical characteristics and designated Aeromonas sp. J-5003. The strain produced two chitinoloytic enzymes: chitinase and chitobiase. The optimum culture conditions of the strain for production of chitinoloytic enzymes were investigated. For the production of chitinase, the major components of medium were colloidal chitin $0.5\%$, glucose $0.2\%$, yeast extract $0.25\%$ and peptone $0.25\%$ while for the production of chitobiase, they were colloidal chitin $0.5\%$, galactose and tryptone $0.2\%$. The optimum cultural temperature and initial pH for the production of chitinase and chitobiase were $30^{\circ}C$ and pH 7.0, respectively.

• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.815-820
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• 2003

A huge database resulted from whole genome sequencing has provided a possibility of new information that is likely to extent the scope and thus changes the way of approach for the functional assigning of putative open reading frames annotated by whole genome sequence analyses. These are mainly realized by ease, one-step identification of putative genes using genomics or proteomics tools. A major challenge remained in biotechnology may translate these informations into better ways to screen or select a gene as a representative sequence. Further attempts to mine the related whole genes or partial DNA fragment from whole genome treasure, and then the incorporation of these sequences into a representative template, will result in the use of putative genes that can be translated into functional proteins or allowed the generation of new lineages as a valuable pool. Such screens enable rapid biochemical analysis and easy isolation of the target activity, thereby accelerating the screening of novel enzymes from the expanded library with related sequences. Information-based PCR amplification of whole genes and reconstitution of functional DNA fragments will provide a platform for expanding the functional spaces of potential enzymes, especially when used mixed- or metagenome as gene resources.

• Proceedings of the Microbiological Society of Korea Conference
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• 2000.10a
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• pp.85-91
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• 2000

Aspergillus oryzae is a filamentous fungus classified in the group Aspergillaceae Ascomycetes. It is an important microorganism for industrial production of enzymes and fermented food productions. It secrets large quantities of proteins or enzymes into the culture medium which makes this organism appealing for the production of heterologous proteins. Recently Electric field-mediated transformation method, electroporation, has been applied to fungal transformation. In this study, fungal transformation was carried out by bypassing the protoplast isolation step, decreasing the culturing time and non-protoplast transformation for the increment of transformation efficiency. Transformants were obtained with electroporation in optimal condition 2,500 voltage, 1,540 ohm and 0.50 capacitance. More than 1,000 transform ants were obtained with 6-10 hrs cultured mycelia without enzyme treatment, called non-protoplast transformation.

• Microbiology and Biotechnology Letters
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• v.46 no.1
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• pp.68-76
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• 2018

Actinobacterial strains isolated from Cow feces were studied for their antifungal attributes against phytopathogens and industrially important enzymes. A total of 30 Actinobacterial strains were obtained from 10 samples of cow feces. All the strains were belonging to the genera Streptomyces on the basis of morphological and chemotaxonomic analysis. During preliminary screening, out of 30 strains, 15 strains (50%) showed antifungal activity against five fungal phytopathogens including Aspergillus niger, Fusarium solani, Fusarium oxysporum, Macrophomina phaseolina and Rhizoctonia solani. While, isolate GBTCF-26 was found to be most active against R. solani with 62.2% inhibition of fungal mycelium, GBTCF-09 was prominent against F. solani and F. oxysporum with percent inhibition of 61.1% and 58.8%, respectively. Out of 30 strains, 19 (63.3%), 16 (53.3%), 11 (36.7%), 10 (33.3%), 4 (13.3%) and 8 (26.7%) strains were producing amylase, caseinase, gelatinase, lipase, chitinase and cellulose, respectively. The selected strains, GBTCF-09, GBTCF-21 and GBTCF-26, were identified as Streptomyces sp. on the basis of their 16S rDNA sequence. The study supports the idea that the Actinobacteria from unique niches (Cow feces) possess the production potential of industrially important enzymes including bioactive molecules.

• Journal of Microbiology and Biotechnology
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• v.3 no.2
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• pp.95-98
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• 1993

This study was carried out to isolate and characterize the mutant strains of Schwanniomyces castellii NRRL Y-2477. Mutants were prepared with the treatment of ethyl methane sulfonate. 2-deoxy-D-glucose resistant mutants were isolated and two mutants were selected based on their high production of amylolytic enzymes and their ability to ferment starch. The mutants selected had higher a-amylase and glucoamylase activities than the wild type strain from several other carbon sources. Especially, it was revealed that mutant strain M-9, when cultured in the presence of glucose as a sole carbon source, shows relatively high activities of a-amylase and glucoamylase compared to those of the wild type strain. In result, this mutant strain can be considered as a constitutive producer of amylolytic enzymes. To compare the ethanol production ability of wild type strain and of mutant strains selected, an alcohol fermentation was carried out using 100 g/l soluble starch. Mutant strain M-9 did not improve the direct alcohol fermentation of starch, despite its excellent amylolytic activities performance. On the other hand, mutant strain M-6 produced 37.9 g/l (4.8%, v/v) ethanol by utilizing about 82% of substrate.

• 한국생물공학회:학술대회논문집
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• 2000.04a
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• pp.215-218
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• 2000

Aspergillus oryzae is a filamentous fungus classified in the group Aspergillaceae Ascomycetes. It is an important microorganism for industrial production of enzymes and fermented food productions. The genetic transformation system in A. oryzae was used to protoplast mediated transformation with PEG/$CaCl_2$. When the protoplast was used, the regeneration efficiency was decreased and then transformation frequence was also effected. In this study, fungal transformation was carried out by bypassing the protoplast isolation step, changing enzymes, such as hemicellulase and celluclast, and decreasing the culturing time for the increment of the transformation efficiency. 83 transformants/10ug of DNA with hemicellulase were obtained, compared with less than 10 transformants with novozyme234 and celluclast.

• Microbiology and Biotechnology Letters
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• v.24 no.2
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• pp.143-148
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• 1996

Thousand actinomycetes and 50 soil samples were used for the isolation of microorganisms producing yeast cell wall lytic enzymes. Among 493 strains producing large clear zones on autolysed washed yeast (AWY), 117 strains were selected on living yeast cell agar plates. With the method of lytic activity, one strain (St-1702) was selected, which was temporarily identified as Streptomyces eurythermus. The optimal condition for enzyme production of this strain was partially determined as follows: incubation of the strain for 3 days at 30$\circ$C in the medium containing 2% freeze dried yeast cell, 1% glucose, 1% K$_{2}$HPO$_{4}$, 0.01% MgSO$_{4}$'7H$_{2}$O, 0.5% peptone, and 0.2% (NH$_{4}$)$_{2}$CO$_{3}$ with pH 7.0. The protoplast formation of yeast by using the enzyme produced by this strain was compared with commercial enzymes.

• Korean Journal of Microbiology
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• v.21 no.4
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• pp.213-220
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• 1983

Conditions for isolation of protoplasts from conidiospores of Trichoderma koningii ATCC 26113 were tested. Maximum production of conidial protoplasts was obtained by preincubation of conidiospores on liquid minimal medium for 8 1/2 hrs. and by reaction with cell wall lytic enzyme for 3 hrs. Among effective cell wall lytic enzymes (Driselase, p-Glucuronidase, Novozyme and Zymolyase), Driselase was the most effective one on the production of conidial protoplasts. The production of conidial protoplasts was also enhanced by addition of 2-Deoxy-D-Glucose $(25{\mu}g/ml)$ into liquid minimal medium. Over 70% of the initial swollen conidia, preincubated in liquid minimal medium supplemented with 2-Deoxy-D-Glucose $(25{\mu}g/ml)$, were converted to protoplasts by incubation with 2% (w/v) commercial lytic enzyme Driselase at $28^{\circ}C$ for 3 hrs. The reversion frequency of the conidial protoplasts was about 30 times (25-50%) higher than that of mycelial protoplasts (0.6-1.3%).

• Fisheries and Aquatic Sciences
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• v.19 no.2
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• pp.9.1-9.7
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• 2016

This study was conducted to identify the general conditions for the isolation and in vitro culture of ovary-derived cells in rockfish (Sebastes schlegeli). The effects of three different enzymes on cell retrieval from ovarian tissues were evaluated first, and then the ovary-dissociated cells were cultured under various culture conditions, with varying basal media and culture temperatures, addition of growth factors, and/or culture types. We found that collagenase type I treatment was effective for cell isolation from ovarian tissues. From a total of 42 trials to evaluate the effects of basal media and culture temperatures on cell culture of ovary-dissociated cells, we observed that Leibovitz's L15 medium was more supportive than Dulbecco's modified Eagle's medium for culture, and the cells could grow at all three temperatures tested, 15, 20, and $25^{\circ}C$, at least up to passage 2. However, growth factor addition did not improve cell growth. Introduction of suspension culture after monolayer culture expanded the culture period significantly more than did monolayer culture alone. Our results may provide a basis for developing an in vitro system for S. schlegeli germline cell culture, which will ultimately lead to improvement of the species.

• 대한생식의학회:학술대회논문집
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• 2003.12a
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• pp.147-148
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• 2003