• BMB Reports
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• v.39 no.1
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• pp.105-110
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• 2006

We have screened 766 strains of fungi from the BIOTEC Culture Collection (BCC) for xylanases working in extreme pH and/or high temperature conditions, the so-called extreme xylanases. From a total number of 32 strains producing extreme xylanases, the strain BCC7928, identified by using the internal transcribed spacer (ITS) sequence of rRNA to be a Marasmius sp., was chosen for further characterization because of its high xylanolytic activity at temperature as high as $90^{\circ}C$. The crude enzyme possessed high thermostability and pH stability. Purification of this xylanase was carried out using an anion exchanger followed by hydrophobic interaction chromatography, yielding the enzyme with >90% homogeneity. The molecular mass of the enzyme was approximately 40 kDa. The purified enzyme retained broad working pH range of 4-8 and optimal temperature of $90^{\circ}C$. When using xylan from birchwood as substrate, it exhibits $K_m$ and $V_{max}$ values of $2.6{\pm}0.6\;mg/ml$ and $428{\pm}26\;U/mg$, respectively. The enzyme rapidly hydrolysed xylans from birchwood, beechwood, and exhibited lower activity on xylan from wheatbran, or celluloses from carboxymethylcellulose and Avicel. The purified enzyme was highly stable at temperature ranges from 50 to $70^{\circ}C$. It retained 84% of its maximal activity after incubation in standard buffer containing 1% xylan substrate at $70^{\circ}C$ for 3 h. This thermostable xylanase should therefore be useful for several industrial applications, such as agricultural, food and biofuel.

• Journal of Environmental Science International
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• v.23 no.6
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• pp.1095-1100
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• 2014

Keratin wastes are generated in excess of million tons per year worldwide and biodegradation of keratin by microorganisms possessing keratinase activity can be used as an alternative tool to prevent environmental pollution. For practical use of keratinase, its physicochemical properties should be investigated in detail. In this study, we investigated characteristics of keratinase produced by Xanthomonas sp. P5 which is isolated from rhizospheric soil of soybean. The level of keratinase produced by the strain P5 increased with time and reached its maximum (10.6 U/ml) at 3 days. The production of soluble protein had the same tendency as the production of keratinase. Optimal temperature and pH of keratinase were $40^{\circ}C-45^{\circ}C$ and pH 9, respectively. The enzyme showed broad temperature and pH stabilities. Thermostability profile showed that the enzyme retained 94.6%-100% of the original activity after 1 h treatment at $10^{\circ}C-40^{\circ}C$. After treatment for 1 h at pH 6-10, 89.2%-100% of the activity was remained. At pH 11, 71.6% of the original activity was retained after 1 h treatment. Although the strain P5 did not degrade human hair, it degraded duck feather and chicken feather. These results indicate that keratinase from Xanthomonas sp. P5 could be not only used to upgrade the nutritional value of feather hydrolysate but also useful in situ biodegradation of feather.

• Journal of Microbiology and Biotechnology
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• v.25 no.5
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• pp.662-671
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• 2015

To enrich the genetic resource of microbial xylanases with high activity and stability under alkaline conditions, a xylanase gene (xynSL4) was cloned from Planococcus sp. SL4, an alkaline xylanase-producing strain isolated from the sediment of soda lake Dabusu. Deduced XynSL4 consists of a putative signal peptide of 29 residues and a catalytic domain (30-380 residues) of glycosyl hydrolase family 10, and shares the highest identity of 77% with a hypothetical protein from Planomicrobium glaciei CHR43. Phylogenetic analysis indicated that deduced XynSL4 is closely related with thermophilic and alkaline xylanases from Geobacillus and Bacillus species. The gene xynSL4 was expressed heterologously in Escherichia coli and the recombinant enzyme showed some superior properties. Purified recombinant XynSL4 (rXynSL4) was highly active and stable over the neutral and alkaline pH range from 6 to 11, with maximum activity at pH 7 and more than 60% activity at pH 11. It had an apparent temperature optimum of 70℃ and retained stable at this temperature in the presence of substrate. rXynSL4 was highly halotolerant, retaining more than 55% activity with 0.25-3.0 M NaCl and was stable at the concentration of NaCl up to 4M. The enzyme activity was significantly enhanced by β-mercaptoethanol and Ca²⁺ but strongly inhibited by heavy-metal ions and SDS. This thermophilic and alkaline- and salt-tolerant enzyme has great potential for basic research and industrial applications.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.3
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• pp.560-565
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• 2004

Bacteriocin producing lactic acid bacteria (LAB) were isolated from Kimchi by using spot-on-the-lawn method. Listeria monocytogenes, Staphylococcus aureus, and Lactobacillus plantarum were used as indicators. One isolate (P3-l) produced a bacteriocin efficiently inhibiting the growth of Listeria monocytogenes. 16S rDNA sequence and sugar utilization test identified that P3-1 was a Lactobacillus sakei strain. Accordingly, the isolate was named as Lactobacillus sakei P3-1. L. sakei P3-1 produced a bacteriocin which efficiently inhibited the growth of Listeria monocytogenes but did not inhibit other Gram positive and negative organisms tested. The bacteriocin was stable against heat, organic solvent, and pH variation and it retained 50% of activity after 10 min heat treatment at 10$0^{\circ}C$. The molecular weight of Sakacin P3-1 was estimated to be 4 kDa by SDS-PAGE.

• Microbiology and Biotechnology Letters
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• v.37 no.4
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• pp.383-388
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• 2009

A strain YB914 was isolated from soil as a producer of the extracellular xylanase, which catalyzes the hydrolysis of oat spelt xylan. The strain YB914 was identified as Streptomyces sp. on the basis of its morphological, cultural and biochemical properties. The xylanase of culture filtrate was the most active at $55^{\circ}C$ and pH 5.5, and retained 80% of its maximum activity at the range of pH 4.5~7.0. In order to optimize the culture medium for xylanase production, ingredients of G.S.S medium were replaced by several carbohydrates. The carbohydrates such as oat spelt xylan, corn cob xylan, wheat bran and lactose increased the xylanase productivity of Streptomyces sp. YB914. However, xylanase production was greatly repressed by galactose or arabinose. The maximum xylanase productivity was reached to 48 U/mL in the modified medium containing 1% oat spelt xylan and 1.5% lactose.

• Mycobiology
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• v.44 no.3
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• pp.121-130
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• 2016

Recently, the Fusarium genus has been narrowed based upon phylogenetic analyses and a Fusarium-like clade was adopted. The few species of the Fusarium-like clade were moved to new, re-installed or existing genera or provisionally retained as "Fusarium." Only a limited number of reference strains and DNA marker sequences are available for this clade and not much is known about its actual species diversity. Here, we report six strains, preserved by the Belgian fungal culture collection BCCM/IHEM as a Fusarium species, that belong to the Fusarium-like clade. They showed a slow growth and produced pionnotes, typical morphological characteristics of many Fusarium-like species. Multilocus sequencing with comparative sequence analyses in GenBank and phylogenetic analyses, using reference sequences of type material, confirmed that they were indeed member of the Fusarium-like clade. One strain was identified as "Fusarium" ciliatum whereas another strain was identified as Fusicolla merismoides. The four remaining strains were shown to represent a unique phylogenetic lineage in the Fusarium-like clade and were also found morphologically distinct from other members of the Fusarium-like clade. Based upon phylogenetic considerations, a new genus, Pseudofusicolla gen. nov., and a new species, Pseudofusicolla belgica sp. nov., were installed for this lineage. A formal description is provided in this study. Additional sampling will be required to gather isolates other than the historical strains presented in the present study as well as to further reveal the actual species diversity in the Fusarium-like clade.

• Microbiology and Biotechnology Letters
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• v.33 no.3
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• pp.178-183
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• 2005

A strain WL-2 was isolated from soil as a producer of the extracellular xylanase, which catalyzes the hydrolysis of oat spelt xylan. The strain WL-2 was identified as Streptomyces sp. on the basis of its 16S rRNA sequence, morphology, cultural and physiological properties. The xylanase of culture filtrate was the most active at $60^{\circ}C$ and pH 6.0, and retained $90{\%}$ of its maximum activity at range of pH $4.5{\~}6.5$. In order to optimize the culture medium for xylanase production, ingredients of G.S.S medium were replaced by several carbohydrates. The carbohydrates such as ${\alpha}-cellulose$, oat spelt xylan and maltose increased dramatically the xylanase productivity of Streptomyces sp. WL-2. The maximum xylanase productivity was reached to 120 U/ml in the modified medium containing $1{\%}\;\alpha-cellulose$ and $1\%}$ maltose.

• The Journal of Advanced Prosthodontics
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• v.11 no.6
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• pp.341-349
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• 2019

PURPOSE. A novel retentive type of implant prosthesis that does not require the use of cement or screw holes has been introduced; however, there are few reports examining the biomechanical aspects of this novel implant. This study aimed to evaluate the biomechanical features of cementless fixation (CLF) implant prostheses. MATERIALS AND METHODS. The test groups of three variations of CLF implant prostheses and a control group of conventional cement-retained (CR) prosthesis were designed three-dimensionally for finite element analysis. The test groups were divided according to the abutment shape and the relining strategy on the inner surface of the implant crown as follows; resin-air hole-full (RAF), resin-air hole (RA), and resin-no air hole (RNA). The von Mises stress and principal stress were used to evaluate the stress values and distributions of the implant components. Contact open values were calculated to analyze the gap formation of the contact surfaces at the abutment-resin and abutment-implant interfaces. The micro-strain values were evaluated for the surrounding bone. RESULTS. Values reflecting the maximum stress on the abutment were as follows (in MPa): RAF, 25.6; RA, 23.4; RNA, 20.0; and CR, 15.8. The value of gap formation was measured from 0.88 to 1.19 ㎛ at the abutment-resin interface and 24.4 to 24.7 ㎛ at the abutment-implant interface. The strain distribution was similar in all cases. CONCLUSION. CLF had no disadvantages in terms of the biomechanical features compared with conventional CR implant prosthesis and could be successfully applied for implant prosthesis.

• Journal of Microbiology and Biotechnology
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• v.33 no.2
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• pp.195-202
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• 2023

Protease is a widely used enzyme particularly in the detergent industry. In this research, we aimed to isolate alkaline protease-producing bacteria for characterization as a laundry detergent additive. The screening of alkaline protease production was investigated on basal medium agar plus 1% skim milk at pH 11, with incubation at 30℃. The highest alkaline protease-producing bacterium was 6BS15-4 strain, identified as Bacillus gibsonii by 16S rRNA gene sequencing. While the optimum pH was 12.0, the strain was stable at pH range 7.0-12.0 when incubated at 45℃ for 60 min. The alkaline protease produced by B. gibsonii 6BS15-4 using dairy effluent was characterized. The optimum temperature was 60℃ and the enzyme was stable at 55℃ when incubated at pH 11.0 for 60 min. Metal ions K⁺, Mg²⁺, Cu²⁺, Na⁺, and Zn²⁺ exhibited a slightly stimulatory effect on enzyme activity. The enzyme retained over 80% of its activity in the presence of Ca²⁺, Ba²⁺, and Mn²⁺. Thiol reagent and ethylenediaminetetraacetic acid did not inhibit the enzyme activity, whereas phenylmethylsulfonyl fluoride significantly inhibited the protease activity. The alkaline protease from B. gibsonii 6BS15-4 demonstrated efficiency in blood stain removal and could therefore be used as a detergent additive, with potential for various other industrial applications.

• Journal of Life Science
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• v.23 no.12
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• pp.1486-1494
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• 2013

A high protease-producing strain was isolated and identified from the digestive tract of octopus vulgaris by detecting a hydrolysis circle of protease and its activity. The strain was identified by morphology observation, biochemical experiments, and 16S rRNA sequence analysis. The protease obtained from the strain was purified by a three-step process involving ammonium sulfate precipitation, carboxy methyl-cellulose (CM-52) cation-exchange chromatography, and DEAE-Sephadex A50 anion-exchange chromatography. The properties of protease were characterized as well. The strain Bacillus sp. QDV-3, which produced the highest activity of protease, was isolated. On the basis of the phenotypic and biochemical characterization and 16S rRNA gene-sequencing studies, the isolate was identified as follows: domain: Bacteria; phylum: Firmicutes; class: Bacilli; order: Bacillales; family: Bacillaceae; and genus: Bacillus. The isolate was shown to have a 99.2% similarity with Bacillus flexus. A high active protease designated as QDV-E, with a molecular weight of 61.6 kDa, was obtained. The enzyme was found to be active in the pH range of 9.0-9.5 and its optimum temperature was $40^{\circ}C$. The protease activity retained more than 96% at the temperature of $50^{\circ}C$ for 60 min. Phenylmethylsulfonyl fluoride (PMSF) inhibited the enzyme activity, thus confirming that this protease isolated from Bacillus sp. QDV-3 is an alkaline serine protease. Metal ions, $Mn^{2+}$ and $Mg^{2+}$, were determined to enhance the protease activity, whereas $Ba^{2+}$, $Zn^{2+}$, and $Cu^{2+}$ were found to inactivate the enzyme.