• Microbiology and Biotechnology Letters
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• v.48 no.1
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• pp.64-71
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• 2020

A total of 262 bacterial strains were isolated from clay minerals, bentonite and zeolite, in Gyeongsangbukdo, Republic of Korea, and their hydrolytic enzyme activities were analyzed. Most of the isolated strains belonged to Micrococcales and Bacillales order. Of strains, 96 strains produced α-amylase activity, 42 strains showed cellulase activity, 111 strains had pectinase activity, and 70 strains showed protease activity. Among them, 177 isolates exhibited one or more of the hydrolytic enzyme activities and in particular Bacillus cereus MBLB1321, B. albus MBLB1326 and KIGAM017, B. mobilis MBLB1328, MBLB1329 and MBLB1330 showed all of the enzyme activities. These results demonstrate the diversity of functional Bacillus species in clay minerals as vital sources for the discovery of industrially valuable hydrolytic enzymes, which have a great commercial prospect in various bio-industrial applications.

• Microbiology and Biotechnology Letters
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• v.25 no.2
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• pp.115-120
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• 1997

Penicillium sp.-L4, a causative fungus of rot in citrus fruits, was isolated and its mode of hydrolytic enzyme production was investigated. Carboxymethylcellulase (CMCase), polygalacturonase(PGase), extra- & intra-cellular $\beta$-glucosidase and cellobiase were produced drastically by addition of substrates in minimal media. Production of the hydrolytic enzymes were induced efficiently by cellobiose and cellooligosaccharides which were the products of cellulose hydrolysis, but repressed by addition of mono-saccharide such as glucose, raffinose, galacturonic acid. The relative activity of p-nitrophenyl-$\beta$-D-glucopyranoside(PNPG) hydrolysis was higher than that of cellobiose hydrolysis in extracellular enzymes, and reverse is true in intracellular enzymes. Intact enzyme production of P. sp.-L4 on lemon peel lesion was sequential. $\beta$-Glucosidase and CMCase were produced first and followed by PGase. The enzyme productivities and pH in lesions were coincident with optimal pH of each enzyme activities.

• Journal of Plant Biotechnology
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• v.6 no.1
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• pp.63-67
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• 2004

A simple procedure for the extraction of the lipolytic enzyme from rice bran has been developed. High activity of lipolytic enzyme was obtained by first defatting the rice bran to remove lipid components with various extraction conditions. Then, after rove cycles of aqueous extraction, rice bran lipolytic enzyme was purified using micro- and ultrafiltration apparatus. Lipolytic enzyme activity was estimated by its hydrolytic action of tributyrin. The result indicated that the standard activity curve of butyric acid showed that the potential rice bran enzyme is a hydrolytic lipase enzyme. In addition, it showed higher lipolytic activity and specific enzyme activity with further purification by micro- and ultrafiltration. The size of rice bran lipase enzyme was identified through 15 ％ SDS-PAGE. The molecular weight of the rice bran lipase enzyme was 41 kDa.

• Journal of the Korean Society of Food Science and Nutrition
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• v.30 no.5
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• pp.839-843
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• 2001

The effect of gamma-irradiation on the hydrolytic enzyme activities of some Koran soybean-based fermented foods was studied. Doenjang (soybean paste), kanjang (soy sauce), kochujang (red pepper paste), chungkukjang and meju were prepared and irradiated at 0, 5, 10 and 20 kGy. Then activities of protease, amylase, lipase and fibrinolytic enzyme were determined. Hydrolytic enzyme activities of meju, chungkukjang and doenjang were relatively higher than those of kanjang and kochujang. Amylase, protease and lipase activities were not affected by 10 kGy and were slightly (about 10%) inactivated by 20 kGy of gamma irradiation, with no statistical significance. Fibrinolytic enzyme was stable in all treatments.

• Proceedings of the KAIS Fall Conference
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• 2004.11a
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• pp.299-301
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• 2004

A simple procedure for the extraction of the lipolytic enzyme from rice bran has been developed. High activity of lipolytic enzyme was obtained by first defatting the rice bran to remove lipid components with various extraction conditions. Then, after five cycles of aqueous extraction, rice bran lipolytic enzyme was purified using micro- and ultrafiltration apparatus. Lipolytic enzyme activity was estimated by its hydrolytic action of tributyrin. The result indicated that the standard activity curve of butyric acid showed that the potential rice bran enzyme is a hydrolytic lipase enzyme. In addition, it showed higher lipolytic activity and specific enzyme activity with further purification by micro- and ultrafiltration.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.5
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• pp.684-689
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• 2006

The present study examined the effects of Tween 80 on the attachment and hydrolytic activity of a cellulase enzyme against ball-milled cellulose (BMC), using the whole component (native CBH I) and the catalysis module (core CBH I) of carbohydrolase I purified from Trichoderma viride (Meicelase, Meiji Seika, Tokyo, Japan). The effects were evaluated as protein concentrations in the supernatant after mixing enzyme and substrate with Tween 80 at room temperature. Tween 80 decreased the adsorption of native CBH I and core CBH I onto BMC (p<0.001) and increased the amount of reducing sugars released from BMC by native CBH I (p<0.001). However, Tween 80 did not enhance the hydrolytic activity of core CBH I. Observations using SEM revealed that Tween 80 caused cellulose filter paper to swell and enhanced surface cracks and filaments caused by native CBH I but not by core CBH I. These results suggested that Tween 80 decreases enzyme adsorption to its substrate but enhances enzymatic activity.

• 한국균학회소식:학술대회논문집
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• 2014.10a
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• pp.47-47
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• 2014

Dokdo island is located in the northeastern part of Ulleungdo, known as volcanic island. In total, 53 fungal isolates were isolated from Dokdo island soil sample, using dilution plate technique. The isolates were identified on the basis of morphological characteristics and rDNA ITS sequence analysis. Out of them, 41 isolates were identified at the level of species. The dominant fungal species and genera included Fusarium spp., Mucor sp., Clonostachys spp., and Trichoderma sp. The % sequence identity (the number of matches/the complete alignment length) values via NCBI BLAST searching of EML-IF9, EML-MF30-1 and EML-DDSF4 represented 97.19% (485/499) with Clonostachys cf. rosea (GenBank accession no. KC313107), 98.33% (472/480) with Metarhizium guizhouense (GenBank accession no. HM055445), and 100% (350/350) with Mortierella oligospora (GenBank accession no. JX976032), respectively. Three species of C. rosea, M. guizhouense and M. oligospora represented new records of fungi from Dokdo island, Korea. The antimicrobial activities of the fungal strains varied with tested. Two isolates (EML-MFS30-1 and EML-IF9) showed antifungal activity against several fungi including Fusarium oxysporum and Rhizotonia solani. Clonostachys rosea (EML-IF9) showed strong hydrolytic enzyme activity. Our results showed that the antagonistic fungi including Clonostachys rosea will be used as potential biocontrol agents for control of fungal diseases.

• 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.

• Journal of the Korean Society of Food Science and Nutrition
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• v.22 no.3
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• pp.273-279
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• 1993

Hydrolytic patterns of 11S globulin (glycinin), storage protein of soybean, by soymilk-clotting enzymes Iand IIfrom Bacillus sp. K-295G-7, which was the first soymilk-clotting enzyme to be found in a bacteria, was investigated. The clotting time of about 4~5 min is revealed by the Enzymes Iand II(0.025 units at 35$^{\circ}C$) on the acidic subunit. In electrophoresis, acidic subunit (A$_3$, M.W. 45,000) disappeared almost completely within 2 min and new products corresponding to the molecular weight of 16,000 and 20,000 were formed by the action of Enzymes I and II. Furthermore, Enzyme II produced a degradation compound having a molecular weight of about 30,000. In contrast, the hydrolytic patterns of basic subunit (M.W. 20,000) by Enzymes I and II were similar, but Enzyme II produced low molecular weight products slower than that of Enzyme I.

• Journal of Microbiology and Biotechnology
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• v.12 no.2
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• pp.189-195
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• 2002

Using Streptomyces sp. YU100 isolated from Korean soil, the fermentative production of phospholipase D was attempted along with its purification and characterization studies. When different carbon and nitrogen sources were supplemented in the culture medium, glucose and yeast extract were found to be the best. By varying the concentration of nutrients and calcium carbonate, the optimal culture medium was determined as 2.0% glucose, 1.5% yeast extract, 0.5% tryptone 0.3% calcium carbonate. During cultivation, the strain secreted most of the phospholipase D in the early stage of growth within 24 h. The phospholipase D produced in the culture broth exhibited hydrolytic activity as well as transphosphatidylation activity on lecithin (phosphatidylcholine). In particular, the culture broth showed 8.7 units/ml of hydrolytic activity when cultivated at $28^{\circ}C$ for 1.5 days. The phospholipase D was purified using 80% ammonium sulfate precipitation and DEAE-Sepharose CL-6B column chromatography, which produced a major band of 57 kDa on a 10% SDS-polyacrylamide gel with purity higher than 80%. The enzyme showed an optimal pH of 7 in hydrolytic reaction, and at pH 4 in a transphosphatidylation reaction. The enzyme activity increased until the reaction temperature was elevated to $60^{\circ}C$. The enzyme was relatively stable at high temperatures and neutral pH, but significantly unstable in the alkaline range. Among the detergents tested as emulsifiers of phospholipids, the highest enzyme activity was observed when 1.5% Triton X-100 was employed. However, no inhibitory effect by metal ions was detected. Under optimized reaction conditions, the purified enzyme not only completely decomposed PC to phosphatidic acid within 1 h, but also exhibited higher than 80% conversion rate of PC to PS by transphosphatidylation within 4 h.