• Title/Summary/Keyword: acid protease A

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Purification and Characterization of an Anticoagulant from Corn Silk (옥발에서 항응고물질의 정제와 특성)

  • Choi, Sang-Kyu;Choi, Hye-Seon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.8
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    • pp.1262-1267
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    • 2004
  • An anticoagulant was purified from corn silk which has been used in Oriental Medicine. The anticoagulant from corn silk has a molecular mass of 135 kDa, and purified by 24 folds with a recovery of 11%. It was not sensitive to heat and protease treatment. However, periodate oxidation of the anticoagulant resulted in loss of activity significantly, implying that a carbohydrate was responsible for an anticoagulant activity. Galactose, glucose, mannose, fucose, glucosamine, and galactosamine were detected after acid hydrolysis by thin layer chromatography (TLC) and Bio-LC. It was confirmed that anticoagulant had OH and NH bonds by IR, supporting that the anticoagulant is composed of neutrosugar and aminosugar. Its anticoagulating activity was measured by delay in thrombin time (TT) and prothrombin time (PT) without affecting clotting by snake venom and delay in activated partial thromboplastin time (APTT). TT was more sensitive than PT, and was delayed two and three times at the concentration of 60 and 88 nM, respectively. The anticoagulating activity was reduced in the thrombin-induced clotting assay using purified fibrinogen according to the increase of fibrinogen concentration with the apparent Ki value of 23 nM.

Production and Action of Microbial Piscicidal Substance (미생물에 의한 살어성물질의 생성 및 그 작용)

  • 도재호;서정훈
    • Microbiology and Biotechnology Letters
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    • v.6 no.1
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    • pp.41-46
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    • 1978
  • Piscicidal substance produced by Streptomyces sp. isolated from soil was toxic against various kinds of fish. After extraction with CH$Cl_3$ from the culture medium, the substance was purified by avicel column chromatography. In order to test toxicity, various kinds of fish were subjected to the acqueous solution of 100 us of the substance per liter of water. Generally, the substance was toxic to most fish, but Macropodus chinenes and Misgurnus mizolepis are resistant to the substance than Gobius similis and Pseudorasbora parva. The substance was stable at pH range, 3.0 to 7.0, but labile at alkaline pH, and to heat as well. Succinic dehydrogenase on most of tissue cell of Cyprinus carpio was inhibited by this substance strongly, but spinal cord was not inhibited. By addition of Cu and Pb salts to the culture medium, piscicidal substance producibility was activated.

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Optimization of Culture Conditions of Bacillus pumilus JB-1 for Chungkook-jang Fermentation in Soybean Boiling-Waste Liquor Medium (대두 열수 침출액을 이용한 청국장 발효균주 Bacillus pumilus JB-1의 배양 최적화)

  • Kwon, Ha-Young;Ryn, Hee-Young;Kwon, Chong-Suk;Lee, Sang-Han;Sohn, Ho-Yong
    • Microbiology and Biotechnology Letters
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    • v.35 no.4
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    • pp.304-309
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    • 2007
  • Soybean is useful source of protein, especially in Asia. But soybean needs heat inactivation or fermentation process before consumption, since it contains the toxic lectin and various protease inhibitors. Therefore, production of soybean boiling-waste liquor (SBWL) as a byproduct is inevitable. In this study, the chemical composition of SBWL and the optimization of culture conditions for Bacillus pumilus JB-1, a selected strain for functional chungkuk-jang fermentation, using SBWL were investigated. The SBWL contains 88% water, 9.5% free sugar, 1.6% crude protein, 0.3% crude fat, 0.1% crude fiber and 2.1% ash, respectively. The contents of total polyphenol, total flavonoids and free amino acid in SBWL were 55%, 76%, and 30% of those of raw soybean, respectively. Culture conditions for B. pumilus JB-1 in SBWL were optimized. The 1/10-diluted, 0.1 % of $(NH_4)_2SO_4$ added SBWL without pH adjustment and carbon source addition was cultured at $37^{\circ}C$ for 48 h with agitation (120 rpm). The 0.5% of inoculation was enough. The large scale fermentation in 5-L jar fermentor showed that the SBWL is a good resource for production of chungkuk-jang starter and functional ingredients.

Quality Characteristics of Doenjang (Soybean Paste) Prepared with Germinated Regular Soybean and Black Soybean (발아 대두 및 검정콩으로 제조한 된장의 품질 특성)

  • Kim, Hyeong-Eun;Han, Song-Yi;Jung, Jin-Bo;Ko, Jong-Min;Kim, Yong-Suk
    • Korean Journal of Food Science and Technology
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    • v.43 no.3
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    • pp.361-368
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    • 2011
  • Three types of doenjang were prepared with regular soybean, germinated regular soybean, and a mixture of germinated regular and black soybean. After 90 days of fermentation, pH, moisture, and salt contents of doenjang were $6.01{\pm}0.01-6.72{\pm}0.01$, $53.78{\pm}0.22-57.99{\pm}0.11$, and $14.17{\pm}0.06-15.03{\pm}0.21%$, respectively, showing slight differences according to soybean germination. Amylase and protease activity of doenjang changed similarly during fermentation in all samples. Amino-type nitrogen and free amino acid content of germinated black soybean doenjang after 90 days of fermentation were $1256.21{\pm}1.86$ and 994.46 mg%, respectively, which were higher than those of other samples. A sensory evaluation of germinated regular soybean doenjang showed the highest scores for color, taste, and overall acceptability. These results suggest that high sensory quality doenjang can be prepared using germinated regular soybean and a mixture of germinated regular soybean and black soybean.

Microbial Diversity in Korean Traditional Fermenting Starter, Nuruk, Collected in 2013 and 2014

  • Seo, Jeong Ah
    • 한국균학회소식:학술대회논문집
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    • 2015.11a
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    • pp.11-11
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    • 2015
  • A total of sixty-six samples of Nuruk, a fermention starter used to make the Korean traditional rice wine, Makgeolli, were collected from central and southern regions of Korea in 2013 and 2014. We classified two groups of the Nuruk samples, "commercial" and "home-made", according to the manufacturing procedure and purpose of use. Commercial Nuruks were made in a controlled environment where the temperature and humidity are fixed and the final product is supplied to Makgeolli manufacturers. Home-made Nuruks were made under uncontrolled conditions in the naturally opened environment and were intended for use in the production of small amounts of home-brewed Makgeolli. We obtained more than five hundred isolates including filamentous fungi and yeasts from the Nuruk samples followed by identification of fungal species. Also we stored glycerol stocks of each single isolate at $-70^{\circ}C$. We identified the species of each isolate based on the sequences of ITS regions amplified with two different universal primer pairs. We also performed morphological characterization of the filamentous fungi and yeast species through observations under the microscope. We investigated the major fungal species of commercial and home-made Nuruks by counting the colony forming units (CFU) and analyzing the occurrence tendency of fungal species. While commercial Nuruks contained mostly high CFU of yeasts, home-made Nuruks showed relatively high occurrence of filamentous fungi. One of the representative Nuruk manufacturers used both domestic wheat bran and imported ones, mainly from US, as raw material. Depending on the source of ingredient, the fungal diversity was somewhat different. Another commercial Nuruk sample was collected twice, once in 2013 and again in 2014, and showed different diversity of fungal species in each year. Nuruks obtained from the southern regions of Korea and Jeju island showed high frequency of yeast such as Saccharomycopsis fibuligera and Pichia species as well as unique filamentous fungus, Monascus species. S. fibuligera was easily found in many Nuruk samples with high CFU. The major filamentous fungi were Aspergillus, Lichtheimia, Mucor and Penicillium species. In order to further our understanding of the isolates and their potential industrial applications, we assayed three enzymes, alpha amylase, glucoamylase and acid protease from 140 isolates out of about five hundred isolates and selected about 10 excellent strains with high enzyme activities. With these fungal isolates, we will perform omics analyses including genomics, transcriptomics, metabolic pathway analyses, and metabolomics followed by whole genome sequencing of unique isolates associated with the basic research of Nuruk and that also has applications in the Makgeolli making process.

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Studies on the N-compounds during Chung-Kook-Jang Meju Fermentation (1) -Changes of Soybean Protein during Chung-Kook-Jang Meju Fermentation- (청국장(淸國醬) 메주 발효과정중(醱酵過程中)의 질소화합물(窒素化合物)의 소장(消長)에 관(關)한 연구(硏究)(I)-대두단백질(大豆蛋白質)의 소장(消長)에 관(關)하여-)

  • Park, Ke-In
    • Applied Biological Chemistry
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    • v.15 no.2
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    • pp.93-109
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    • 1972
  • Three lots of Chung-Kook-Jang were prepared by the use of 2 strains of Bacillus subtilis and Bacillus natto. For four samples taken from each lot in 12 hrs interval changes of nitrogenous compounds, insoluble protein, water soluble protein, peptides, free amino acids, amino and ammonia nitrogens during Chung-Kook-Jang fermentation, were studied together with the changes of moisture, pH, proteolytic enzyme activity. In addition the average peptide length of the peptides of a Bacillus subtilis lot was determined by the method of molecular sieving using ion exchange resin. The results were as follows: 1. The contents of moisture and total-nitrogen changed little in all samples throughout the fermentation as it would be expected. 2. In all three experimental lots the pH became higher gradually from the initial value of 6.65 to the final $7.5{\sim}7.85$ during the fermentation. Proteolytic enzyme activities, in accordance with this pH change, steadily increased up to $48{\sim}60$ hrs. of fermentation and then slightly decreased, probably affected by the high pH. The most strong proteolytic activity was observed in the experimental Chung-Kook-Jang fermentation lot using the Bacillus subtilis K-27 isolated by the author. 3. The contents of insoluble protein nitrogen in soybeans increased markedly (5%) by the cooking, after steeping 12 hrs in water. During the Chung-Kook-Jang fermentation, however, it decreased from 1/2 to 1/10 of that of the cooked soybeans. 4. The contents of water soluble protein nitrogen (5%) whereas, greatly decreased to the value of 1.0% by the cooking; but little changed further during the fermentation, 5. The total contents (0.25%) of peptides, amino, and ammonia-nitrogens, PAA-N., increased almost double by the cooking and steadily became higher as the fermentation proceeded, reaching finally up to$4{\sim}7%$ in 72 hrs fermentation. 6. The amounts of free amino acids of soybean generally decreased during the processing of cooking, even some of them like glutamic acid were destroyed completely, However in the subsequent Chung-Kook-Jang fermentation for 72 hrs., they showed from several to a few hundreds folds increases depending upon the kinds of amino acids. Valine which was contained in HCl-hydrolyzed steeped or cooked soybeans in amounts $220{\sim}267mg%$ was not detected at all as the free amino acid in all fermented samples. 7. Average peptide length (APL) of all fractions, eluted and fractionated by using the Dowex-50 ion exchange resin column, and fraction collector showed the highest value for the cooked soybean and then decreased as the fermentation proceeded. The APL value of effluent showed the highest in 12 hrs fermented sample, The value decreased thereafter by fermentation.

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Optimization of enzymatic hydrolysis of viscera waste proteins of black body fowl(Yeonsan Ogae) to produce peptides using a commercial protease and it's characters analysis (단백질 분해효소를 이용한 연산오계 내장 펩타이드 생산 최적화 및 특성분석)

  • Choi, So-Young;Kim, A-Yeon;Song, Yu-Rim;Ji, Joong-Gu;Yoo, Sun-Kyun
    • Journal of Digital Convergence
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    • v.14 no.1
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    • pp.253-262
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    • 2016
  • Yeonsan Ogae has been known as supporting health and high efficacy of treatment. In recent days, as the efficacy of functional peptides has known, the optimization of oligo peptides production and its characteristics from Ogae viscera has been performed. Response surface method was used to perform the optimizaion of enzyme hydrolysis. The range of processes was temperature (40, 50 and $60^{\circ}C$), pH(6.0, 7.0 and 8.0), and enzyme(1, 2 and 3%). The degree of hydrolysis, amono acids, molecular weight of products were analyzed. The optimum process of enzyme hydrolysis were determined as temperature $58^{\circ}C$, pH 7.5, and enzyme concetration 3%. At optimum conditions, the degree of hydrolysis after 2 h reaction was 75-80%. The total amino acids of amino acid and were 386.15 mg/100 g and 155.26 mg/100 g, respectively. The molecular weight of products by using Maldi-TOF was ranged from 300 to 1,000 Da.

Comparison for enzymic activity of Nuruk and quality properties of Yakju by different fungi (곰팡이 균주에 따른 누룩의 효소활성 및 약주 품질특성 비교)

  • Huh, Chang-Ki;Kim, So-Mang;Kim, Yong-Doo
    • Food Science and Preservation
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    • v.21 no.4
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    • pp.573-580
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    • 2014
  • The enzymatic activity of Nuruk and the quality properties of Yakju were investigated according to different fungi. The fungi that were used in this study were Aspergillus kawachii KCCM 32819, Aspergillus niger KCCM 32005, Rhizopus japonicus KCCM 11604, Rhizopus oryzae KCCM 11272, Rhizopus oryzae KCCM 11273, Rhizopus oryzae KCCM 11276, and Mucor rouxii KCCM 60148. The study results are as follows. The saccharogenic power of Rhizopus oryzae KCCM 11272 Nuruk was the highest (3,647.72 SP/g) among all the samples. The ${\alpha}$-amylase production and protease activities were highest (3.76 DU and 4.7 tyrosine mg/min, respectively) in the Rhizopus japonicus KCCM 11604 Nuruk. The pH levels of the Yakju made with commercial Nuruk and Rhizopus japonicus KCCM 11604 Nuruk were 4.14 and 4.07, respectively. The total titratable acid content of the Yakju made with Rhizopus oryzae KCCM 11273 Nuruk was the highest (0.56%) among all the samples. Rhizopus japonicus KCCM 11604 and Rhizopus oryzae KCCM 11272 had the highest ethanol yields (15.18% and 15.10%, respectively). In the sensory evaluation carried out in this study, the panel preferred the Yakju made with Rhizopus japonicus KCCM 11604 Nuruk. Overall, however, the panel did not like the Yakju made with Aspergillus niger KCCM 32005 Nuruk.

Structure-Function Analysis of DNA Binding Domain of the Yeast ABF1 Protein (효모 ABF1 단백질의 DNA Binding 부위에 대한 구조 기능 연구)

  • Cho, Gi-Nam;Lee, Sang-Kyung;Kim, Hong-Tae;Kim, Ji-Young;Rho, Hyune-Mo;Jung, Gu-Hung
    • Korean Journal of Microbiology
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    • v.32 no.2
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    • pp.102-108
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    • 1994
  • Autonomously replicating sequence Binding Factor 1(ABF1) is a DNA-binding protein that specifically recognizes the $RTCRYN_5ACG$ at many sites in the yeast genome including the promoter element, mating-type silencer and ARS. To express the intact full-length ABF1 gene in E. coli, the ABF1 gene has been cloned into pMAL-c2 and His-61, Leu-353 and Leu-360 were substituted with other amino acid. ABF1 fusion proteins of wild type ABF1 and H61A, L353R and L360R nutants were purified by amylose resin affinity chromatography. Fusion protein of MBP and ABF1 was digested by Factor Xa and Characterized by gel retardation assay and complementation test. As aresult, we suggested that other DNA binding motif except atypical inc-finger motif is in the middle region of ABF1.

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EFFECT OF CHLORHEXIDINE ON MICROTENSILE BOND STRENGTH OF DENTIN BONDING SYSTEMS (Chlorhexidine 처리가 상아질 접착제의 미세인장결합강도에 미치는 영향)

  • Oh, Eun-Hwa;Choi, Kyoung-Kyu;Kim, Jong-Ryul;Park, Sang-Jin
    • Restorative Dentistry and Endodontics
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    • v.33 no.2
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    • pp.148-161
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    • 2008
  • The purpose of this study was to evaluate the effect of chlorhexidine (CHX) on microtensile bond strength (${\mu}TBS$) of dentin bonding systems. Dentin collagenolytic and gelatinolytic activities can be suppressed by protease inhibitors, indicating that MMPs (Matrix metalloproteinases) inhibition could be beneficial in the preservation of hybrid layers. Chlorhexidine (CHX) is known as an inhibitor of MMPs activity in vitro. The experiment was proceeded as follows: At first, flat occlusal surfaces were prepared on mid-coronal dentin of extracted third molars. GI (Glass Ionomer) group was treated with dentin conditioner, and then, applied with 2 % CHX. Both SM (Scotchbond Multipurpose) and SB (Single Bond) group were applied with CHX after acid-etched with 37% phosphoric acid. TS (Clearfil Tri-S) group was applied with CHX, and then, with adhesives. Hybrid composite Z-250 and resin-modified glass ionomer Fuji-II LC was built up on experimental dentin surfaces. Half of them were subjected to 10,000 thermocycle, while the others were tested immediately. With the resulting data, statistically two-way ANOVA was performed to assess the ${\mu}TBS$ before and after thermo cycling and the effect of CHX. All statistical tests were carried out at the 95 % level of confidence. The failure mode of the testing samples was observed under a scanning electron microscopy (SEM). Within limited results, the results of this study were as follows; 1. In all experimental groups applied with 2 % chlorhexidine, the microtensile bond strength increased, and thermo cycling decreased the micro tensile bond strength (P > 0.05). 2. Compared to the thermocycling groups without chlorhexidine, those with both thermocycling and chlorhexidine showed higher microtensile bond strength, and there was significant difference especially in GI and TS groups. 3. SEM analysis of failure mode distribution revealed the adhesive failure at hybrid layer in most of the specimen. and the shift of the failure site from bottom to top of the hybrid layer with chlorhexidine groups. 2 % chlorhexidine application after acid-etching proved to preserve the durability of the hybrid layer and microtensile bond strength of dentin bonding systems.