• 한국생물공학회:학술대회논문집
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• 2000.04a
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• pp.527-530
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• 2000

A kiwi fruit ,called as the Chinese gooseberry, is originated from the Yangtze River Valley of Northern China and Zhejiang Province on the cost of Eastern China. Around 1950, a large mass production began at New Zealand with an Improved breeding. Plant origin actinidin from kiwi fruit belongs to the papain family of cysteine proteinase, which in plants includes papain from papaya, bromelain from pineapple, Cl4 protease from tomato and aleurain from barley. Actinidin is involved in the ripening-related gene family. In this study, protease gene of chinese wild kiwi fruit was isolated and characterized. 1.2kb PCR-amplified fragment was obtained from the total RNA using RT-PCR. pWACT-1 was obtained by subcloning of amplified fragment into pGEM-T Easy cloning vector and analyzed nucleotide sequence by DNA sequencing and amino acid sequence. In Result, high levels of homology between wild kiwi and New Zealand cultured-kiwi was obtained.

• Mycobiology
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• v.42 no.3
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• pp.291-295
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• 2014

Mycoflora was assessed in the commercial meju from four well-separated geographic origins. A total of 112 fungal isolates were identified by phenotypic characteristics and molecular taxonomy using sequencing the internal transcribed spacer of the rDNA and revealed 19 species from 13 genera. Enzymatic characteristics of protease and amylase, and mycotoxin production were analyzed.

• 한국생물공학회:학술대회논문집
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• 2002.04a
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• pp.427-430
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• 2002

Pilot scale(2.5 ton) three-stage methane fermentation process was developed for the rapid production of methane from food wastes in our laboratory. Eleven strains responsible for the primary semianaerobic hydrolysis/acidogenic fermentation system were isolated and characterized. Among them, the number of gram positive bacteria was eight and that of gram negative bacteria was three. They were rod and showed positive reaction to catalase. The strain K5 was found to have the highest enzyme activities of amylase and protease.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1998.11a
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• pp.161-161
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• 1998

To achieve the aim of this investigation, the extracellular protease was isolated from bacteria BAC-4, a strain was cultivated in the medium for the production of penicillin acilase in a period of 32 hours. The enzyme was first purified by aceton precipitation method, followed by ion exchange chromatography on DEAE-sephacel column. The highest specific activity of the aceton fraction was found to be 2.19 unit per mg, with degree of purification of 13 times. Further purification of the enzyme on DEAE -sephacel had a specific activity of 58.6 unit per mg and degree of purification of 344 times compared to its crude extract. The optimum pH of the enzyme was 8.4, and the potimum temparature was 37$^{\circ}C$. The K$\_$M/ and $V_{max}$ calculated at experiment conditions were found to be 0.66%(W/V) and 3.61 unit per mL respectively.

• The Korean Journal of Mycology
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• v.51 no.3
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• pp.219-227
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• 2023

This study was conducted to analyze the distribution and characteristics of fungal species in meju using the traditional method. Fungal distribution in meju was investigated using metagenomic and morphological analyses, based on which Aspergillus flavus/oryzae strains were identified as the dominant fungi in all meju samples, followed by Pichia, Rhizopus and Lichtheimia spp. As A. flavus/oryzae was dominant, we further evaluated the aflatoxin production ability and enzymatic activity of the isolates. Thin-layer chromatography and polymerase chain reaction revealed that the A. flavus/oryzae strains isolated from meju are non-aflatoxigenic fungi. Based on the analyses of amylase and protease activities, strains with high activities of amylase or protease were identified, which are proposed to be used as starters for meju fermentation.

• Korean Journal of Microbiology
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• v.37 no.3
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• pp.214-220
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• 2001

To develop the effective composting system, we isolated bacteria that have the abilities to degrade organic matters such as cellulose, carbohydrate, protein and lipid during the compositing of livestock manureAmong 24 strains, 6 bacteria have all the enzymatic activities of protease, amylase, cellulase and lipase.These microorganisms were identified as Corynebacterium varibilis, Bacillus spp., Pseudomonas spinosa,Acetobacter calcoaceticus and Athrobacter cumminsii All the enzymes produced by the bacteria showedactivities at the broad pH range and the maximal activities were obtained at $60^{\circ}C$. It seemed that after theincrease of temperature caused by fermentation of livestock manure, the enzymes started to degrade the rawmaterials, which are added for the control of humidity. However cellulase activity was maximum at $37^{\circ}C$,suggesting that the cellulase-producing bacteria work at an early stage of livestock manure fermentation toprovide the organic material for the growth of other bacteria. The production of the enzymes were growth-associated and maximal activities appeared at the early stationary phase of growth.

• Applied Biological Chemistry
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• v.41 no.1
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• pp.73-77
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• 1998

The effects of soymilk residues solubilization by cellulase, protease, koji and yeast were investigated on dry matter and protein yields, amino acid and organic acid contents. Co-treatment of soymilk residues by cellulase and protease gave high dry matter yield and protein yield. Koji treatment followed by yeast fermentation was effective for increasing organic acid content and producing soy sauce-like taste and odor. Organic acid content of fermented hydrolysates was improved by cellulase treatment. Protease treatment rather than koji treatment gave high amino acid content, and cellulase treatment seemed to have little effect on increasing free amino acid content. In sensory evaluation, koji-treated hydrolysate showed higher overall acceptance than other hydrolysates, however it showed lower overall acceptance than commercial fermented soy sauce.

• KSBB Journal
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• v.31 no.4
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• pp.284-290
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• 2016

In this research, recombinant human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) was produced by transgenic rice cells. RAmy3D promoter was used for overcome the limitation of low expression level in transgenic plant cells, and the secretion of target protein was accomplished by signal peptide. However, the RAmy3D promoter system which can be induced only by sugar starvation causes the decrease of cell viability. As a result, cell death promotes the release of protease which degrades the target proteins. The protein stability and productivity can be significantly influenced by proteolysis activity. Therefore, development of new strategies are necessary for the in situ recovery of target proteins from cell culture media. In this study, in situ recovery was performed by various strategies. Direct addition of Protein A resin with nylon bag leads to cell death by increased shear stress and decrease in production of hCTLA4Ig by protease. Medium exchange through modified flask could recover hCTLA4Ig with high cell viability and low protease activity, on the other hand, the productivity was lower than that of control. When in situ recovery was conducted at day 7 after induction in air-lift bioreactor, 1.94-fold of hCTLA4Ig could be recovered compared to control culture without in situ recovery. Consequently, in situ recovery of hCTLA4Ig from transgenic rice cell culture could enhance productivity significantly and prevent degradation of target proteins effectively.

• Korean Journal of Food Science and Technology
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• v.33 no.6
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• pp.769-773
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• 2001

Protein content of okara and soybean were found to be 37.3% and 42.5%, respectively by micro-Kjeldahl analysis. Solubility of okara protein in phosphate buffer (pH 8) was 10% versus soy protein of 68.4%. Insolubilization of okara protein was mostly due to disulfide bonding between cysteine residues caused by excessive heat treatment during soymilk processing: hydrophobic interactions and hydrogen bondings were involved to lesser extent. Optimum extraction temperature and time were $60^{\circ}C$ and 40 min. Typical solubility profile of soy protein disappeared for okara protein though minimum solubility of the protein was around pH 3.0. Treating okara with protease was effective in solubilizing okara protein and solubility increased to 19.2%. Optimum reaction temperature and time were $80^{\circ}C$ and 50 min, respectively. Cell wall degrading enzyme did not increase solubility of the protein, however. Through enzymatic reaction okara protein could be effectively solubilized for uses as food ingredient.

• Fisheries and Aquatic Sciences
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• v.7 no.1
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• pp.10-15
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• 2004

A total of 52 pathogenic Vibrio strains was isolated from the Gwangan Beach during summer in 2003. The isolated vibrios were composed of 6 different species: V. parahaemolyticus, V. cholerae non O1, V. fluvialis, V. vulnificus, V. alginolyticus, and V. mimicus. V. parahaemolyticus was most predominant as $46\%$ (24/52), V. cholerae non O1 was the second with $23\%$ (12/52), and V. fluvialis was the third with $17\%$ (9/52). Among the isolated strains, 22 strains showed hemolytic, proteolytic or ureolytic activity. Eight strains showed both hemolysin and protease activities, and either 6 strains showed only hemolysin activities and 7 strains only protease activities. Only one strain of V. parahaemolyticus isolates showed urease activity. The urease-positive V. parahaemolyticus strain (V. parahaemolyticus S25) showed the same biochemical characteristics as the reference strain, V. parahaemolyticus KCTC 2471 (urease­negative) except for urease production. To compare the degree of virulence of Vibrio strains having different pathogenic factors, hemolysin, protease, or urease-positive strains were injected into groups of 10 each of ICR mice (7- to l0-week-old male). The lethal rate of urease-positive V. parahaemolyticus S25 was significantly high, being $70\%$. Protease-positive strains showed $40-60\%$ of lethal rate. Hemolysin-positive strains showed no mortality, similar to non-pathogenic V. parahaemolyticus KCTC 2471 and V. parahaemolyticus FM12.