• The Korean Journal of Mycology
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• v.43 no.4
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• pp.239-246
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• 2015

Two fungal strains were isolated from rods of Quercus sp. (NAAS02335) and Pinus densiflora (NAAS05299) in Korea. These strains were identified as Gyrodontium sacchari by their morphological and mycological characteristics. The optimal growth temperature of NAAS02335 and NAAS05299 are $25^{\circ}C$ and $30^{\circ}C$, respectively. Production of cellulase, xylanase, and ligninase was tested on agar media supplemented dyes or substrates. Production of cellulase and xylanase of NAAS05299 was higher than those of NAAS02335, however ligninase activity of NAAS02335 was higher than that of NAAS05299. The activities of cellulase, xylanase, and amylase of strain NAAS05299 were estimated at 6.7~10.2 times higher than that of NAAS02335. Laccase activity was only estimated by strain NAAS02335. The lignocellulytic enzymes are induced by substrates such as rice straw, wooden chips of pine, oak, and poplar. The NAAS05299 was able to degrade filter paper completely after 4 weeks of culturing in liquid media containing a piece of filter paper at $28^{\circ}C$ with continuous shaking. NAAS05299 was able to degrade rice straw, pine chips, and oak chips after 4 months in solid culture, however NAAS02335 decomposed only rice straw among tested 4 kinds of biomass.

• Microbiology and Biotechnology Letters
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• v.16 no.4
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• pp.275-281
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• 1988

To develop cellulase and xylanase-producing strain by protoplast fusion, alkalophilic Bacillus sp. F204 and K17 were treated with NTG(N-methyl-N'-nitro-N-nitrosoguanidine) and isolated anti-biotics resistant strains of S20 (Km$^r$ , Cm$^r$) and G70 (Str$^r$). The frequency of protoplast formation was about 95% when cells of mid-log phase were treated with 200$\mu\textrm{g}$/ml Iysozyme at 37$^{\circ}C$ for 30-45 minutes. Under addition of 0.4-0.5M sodium succinate, 0.5% casamino acid, 1.5% polyvinylpyrrolidone, 25mM MgC1$_2$ and 50mM CaC1$_2$ to the regeneration medium, the regeneration frequency of Bacillus sp. F204 and K17 was 24.9% and 26.2%, respectively. The fusion frequency was 6.6$\times$10$^{-6}$ in the presence of 30% polyethylene glycol 6000 containing 50mM $Ca^{++}$ at 45$^{\circ}C$ for 5 minutes. Cellulase complex and xylanase activities of fusant were compared with parental strains.

• Journal of Microbiology and Biotechnology
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• v.21 no.12
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• pp.1322-1329
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• 2011

Filamentous fungi colonizing rice straw were collected from 11 different sites in Korea and were identified based on characterization of their morphology and molecular properties. The fungi were divided into 25 species belonging to 16 genera, including 14 ascomycetes, one zygomycete, and one basidiomycete. Fungal cellulolytic and xylanolytic enzymes were assessed through a two-step process, wherein highly active cellulase- and/or hemicellulase-producing fungi were selected in a first screening step followed by a second step to isolate the best enzyme-producer. Twenty-five fungal species were first screened for the production of total cellulase (TC), endo-${\beta}$-1,4 glucanase (EG), and endo-${\beta}$-1,4 xylanase (XYL) using solid-state fermentation with rice straw as substrate. From this screening, six species, namely, Aspergillus niger KUC5183, A. ochraceus KUC5204, A. versicolor KUC5201, Mucor circinelloides KUC6014, Trichoderma harzianum 1 KUC5182, and an unknown basidiomycete species, KUC8721, were selected. These six species were then incubated in liquid Mandels' media containing cellulose, glucose, rice straw, or xylan as the sole carbon source and the activities of six different enzymes were measured. Enzyme production was highly influenced by media conditions and in some cases significantly increased. Through this screening process, Trichoderma harzianum 1 KUC5182 was selected as the best enzyme producer. Rice straw and xylan were good carbon sources for the screening of cellulolytic and xylanolytic enzymes.

• The Korean Journal of Mycology
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• v.26 no.1 s.84
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• pp.127-133
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• 1998

Two regulatory mutants of Trichoderma reesei QM 9414 were isolated by treatment with N-methyl-N'-nitro-N-nitrosoguanidine, and the effects of various inducers on the carboxymethyIcellulose (CMC) and filter paper (FP) production were investigated. Induction of CMCase and FPase production of mutants was shown higher level than wild type strain in 1% lactose minimal broth. When induced by glucose, wild type showed glucose-repression for CMCase and FPase production and mutants showed glucose-derepression. Mutant 1 showed 8.38 fold higher CMCase activity and 5.68 fold higher FPase activity than wild type stain. Mutant 2 showed about 8.42 fold higher CMCase activity and 5.41 fold higher FPase activity than wild type strain. Enzyme activities from the mutants and wild type had the same optimum pH of 4.8 and optimum temperature of $60^{\circ}C$.

• Microbiology and Biotechnology Letters
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• v.47 no.1
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• pp.125-131
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• 2019

We have isolated and identified 72 bacterial strains from four bentonite samples collected at the mining areas located in Gyeongsangbuk-do, Republic of Korea, and measured their hydrolytic enzyme (${\alpha}$-amylase, protease, and cellulase) activities to identify the isolates with industrial-use potential. Most of the isolates belonged to the Bacillaceae, with minor portions being from the Paenibacillaceae, Micrococcaceae, and Bacillales Family XII at the family level. Of the strains isolated, 33 had extracellular ${\alpha}$-amylase activity, 30 strains produced cellulase, and 35 strains produced protease. Strain MBLB1268, having the highest ${\alpha}$-amylase activity, was identified as Bacillus siamensis ($0.38{\pm}0.06U/ml$). Bacillus tequilensis MBLB1223, isolated from Byi33-b, showed the highest cellulase activity ($0.26{\pm} 0.04U/ml$), whereas Bacillus wiedmannii MBLB1197, isolated from Zdb130-b, exhibited the highest protease activity ($54.99{\pm}0.78U/ml$). These findings show that diverse bacteria of the Bacillaceae family adhere to and exist in bentonite and are potential sources of industrially useful hydrolytic enzymes.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.529-532
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• 2001

Korean food waste, one of the abundantly available but environmentally problematic organic wastes in Korea, was utilized as solid-substrate by fungal strain Aspergillus niger ATcC 6275 for the production of enzymemixture containing amylase, cellulase and xylanase. The enzyme mixture can be used as high value-added animal feed. Solid-state fermentation method yielded a 84-fold enhancement in xylanase activity compared with submerged fermentation method. The effect of incubation period, incubation temperature, pH of medium, moisture content, inoculum size and enrichment of the medium with nitrogen and carbon sources were observed for optimal production of these enzymes The optimal amylase activity of 33.10 U/g, cellulase activity of 24.41 U/g, xylanase activity of 328.84 U/g were obtained at 8 days incubation with 50%(w/w) soy bean flake, with incubation temperature of $25^{\circ}C$, pH of 6.38, optimal moisture content of 55% and with inoculum size of $3.8{\times}10^6$spore/g. Enzyme activities were enhanced when ImM $CaSO_4$, 2% Malt extract and 2% galactose were added as mineral, nitrogen and carbon enrichment respectively.

• Horticultural Science & Technology
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• v.19 no.4
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• pp.545-549
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• 2001

The relationship between ethylene action and flesh softening in post-storage persimmon fruits was investigated by treating the fruits with 1-MCP. The patterns of firmness change caused by various treatments with 1-MCP and ethylene were similar to those of changes in cell wall-degrading enzyme activities, including cellulase, PG, PME, and ${\beta}$-galactosidase. Moreover, the activities of these enzymes were inhibited by 1-MCP treatment. These results show that the cell wall-degrading enzymes influenced by ethylene account for the flesh softening process of post-storage persimmon fruit. The ethylene production of fruits, as measured by ACC content and ACC oxidase activity, nevertheless, was not influenced by 1-MCP treatments. It is suggested that the flesh softening phenomena in post-storage persimmon fruits is correlated to the ethylene responsiveness of tissue rather than the ethylene production rate per se.

• Korean Journal of Microbiology
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• v.47 no.2
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• pp.130-136
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• 2011

Five strains of Actinomyces were isolated from freshwater sponges, Baikalospongia and Lubomirskia, in Lake Baikal. By 16S rRNA sequencing, isolates were identified as Streptomyces griseoplanus, S. halstedii, S. violascens, S. flavovirens, and S. microflavus. Isolates had different characteristics of growth temperature, carbon utilization, enzyme activity, and cellular fatty acid composition. Optimum growth conditions of isolates were $30-37^{\circ}C$, pH 8-9, and 0-1.5% salt concentrations. Major fatty acid compositions were anteiso-$C_{15:0}$, iso-$C_{15:0}$, and iso-$C_{16:0}$. Strain ATS-BA-19 had DNase and chitinase activities and strain ATS-BA-16 had cellulase and protease activities. Colonies of strain ATS-BA-15 and ATS-BA-19 made inhibition zone of Pseudomonas aeruginosa.

• Journal of Microbiology and Biotechnology
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• v.24 no.10
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• pp.1427-1437
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• 2014

Enzymatic hydrolysis of lignocellulosic biomass into fermentable sugars is a key step in the conversion of agricultural by-products to biofuels and value-added chemicals. Utilization of a robust microorganism for on-site production of biomass-degrading enzymes has gained increasing interest as an economical approach for supplying enzymes to biorefinery processes. In this study, production of multi-polysaccharide-degrading enzymes from Aspergillus aculeatus BCC199 by solid-state fermentation was improved through the statistical design approach. Among the operational parameters, yeast extract and soybean meal as well as the nonionic surfactant Tween 20 and initial pH were found as key parameters for maximizing production of cellulolytic and hemicellulolytic enzymes. Under the optimized condition, the production of FPase, endoglucanase, ${\beta}$-glucosidase, xylanase, and ${\beta}$-xylosidase was achieved at 23, 663, 88, 1,633, and 90 units/g of dry substrate, respectively. The multi-enzyme extract was highly efficient in the saccharification of alkaline-pretreated rice straw, corn cob, and corn stover. In comparison with commercial cellulase preparations, the BCC199 enzyme mixture was able to produce remarkable yields of glucose and xylose, as it contained higher relative activities of ${\beta}$-glucosidase and core hemicellulases (xylanase and ${\beta}$-xylosidase). These results suggested that the crude enzyme extract from A. aculeatus BCC199 possesses balanced cellulolytic and xylanolytic activities required for the efficient saccharification of lignocellulosic biomass feedstocks, and supplementation of external ${\beta}$-glucosidase or xylanase was dispensable. The work thus demonstrates the high potential of A. aculeatus BCC199 as a promising producer of lignocellulose-degrading enzymes for the biomass conversion industry.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.3
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• pp.374-379
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• 2003

Four rumen fistulated Murrah buffaloes were used to study the effect of four diets differing in roughage to concentrate ratio on rumen biochemical changes, microbial enzyme profile and in sacco degradability of feed in a $4{\times}4$ Latin Square design. The animals were fed four diets consisting of 80:20, 70:30, 60:40 and 50:50 ratios of wheat straw and concentrate mixtures, respectively. Wheat straw and concentrate mixture were mixed with water (0.6 l/kg feed) and complete feed mixture was offered to the animals at 8:00 h and 16:00 h in two equal parts. The variation in pH of rumen liquor (difference of maximum and minimum during 0-8 h post feeding) increased with increasing level of concentrate mixture in the diet. There was no effect of diet composition on volatile fatty acids, total nitrogen and trichloro-acetic acid precipitable nitrogen in the rumen liquor, but ammonia nitrogen increased with increasing level of concentrate mixture in the ration. Major portions of all fibre degrading enzymes were present in the particulate material (PM) of the rumen contents, but protease was absent in PM fraction. The activities of micro-crystalline cellulase, acetyl esterase and protease increased with increase in the level of concentrate mixture, but the activities of other enzymes (carboxymethylcellulase, filter paper degrading activity, xylanase, $\beta$-glucosidase and $\beta$-xylosidase) were not affected. The in sacco degradability and effective degradability of feeds increased with increasing level of concentrate mixture in the ration.