• Title/Summary/Keyword: Acid protease

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Isolation and Characterization of Various Strains of Bacillus sp. having Antagonistic Effect Against Phytopathogenic Fungi (식물 병원성 곰팡이에 길항작용을 갖는 다양한 Bacillus sp.의 균주 분리와 특성에 관한 연구)

  • Kim, Hee Sook;Kim, Ji-Youn;Lee, Song Min;Park, Hye-Jung;Lee, Sang-Hyeon;Jang, Jeong Su;Lee, Mun Hyon
    • Microbiology and Biotechnology Letters
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    • v.47 no.4
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    • pp.603-613
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    • 2019
  • This study was carried out to examine the antagonistic effect against phytopathogenic fungi of isolated strains from soil samples collected from Busan, Changwon, and Jeju Island: Botrytis cinerea, Colletotrichum acutatum, Corynespora cassiicola, Fusarium sp., Rhizoctonia solani, Phytophthora capsici, and Sclerotinia sclerotiorum. According to results of our studies, isolated strains showed an antagonistic effect against phytopathogenic fungi. Such an antagonistic effect against phytopathogenic fungi is seen due to the production of siderophores, antibiotic substances, and extracellular amylase, cellulase, protease, and xylanase enzyme activities. Extracellular enzymes produced by isolated strains were significant, given that they inhibited the growth of phytopathogenic fungi by causing bacteriolysis of the cell wall of plant pathogenic fungi. This is essential to break down the cell wall of plant pathogenic fungi and thus help plant growth by converting macromolecules, which cannot be used by the plant for growth, into small molecules. In addition, they are putative candidates as biological agents to promote plant growth and inhibit growth of phytopathogenic fungi through nitrogen fixation, indole-3-acetic acid production, siderophore production, and extracellular enzyme activity. Therefore, this study suggests the possibility of using Bacillus subtilis ANGa5, Bacillus aerius ANGa25, and Bacillus methylotrophicus ANGa27 as new biological agents, and it is considered that further studies are necessary to prove their effect as novel biological agents by standardization of formulation and optimization of selected effective microorganisms, determination of their preservation period, and crop cultivation tests.

Intraspecies Volatile Interactions Affect Growth Rates and Exometabolomes in Aspergillus oryzae KCCM 60345

  • Singh, Digar;Lee, Choong Hwan
    • Journal of Microbiology and Biotechnology
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    • v.28 no.2
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    • pp.199-209
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    • 2018
  • Volatile organic compounds (VOCs) are increasingly been recognized as the chemical mediators of mold interactions, shaping their community dynamics, growth, and metabolism. Herein, we selectively examined the time-correlated (0 D-11 D, where D = incubation days) effects of intraspecies VOC-mediated interactions (VMI) on Aspergillus oryzae KCCM 60345 (S1), following co-cultivation with partner strain A. oryzae KACC 44967 (S2), in a specially designed twin plate assembly. The comparative evaluation of $S1_{VMI}$ (S1 subjected to VMI with S2) and its control ($S1_{Con}$) showed a notable disparity in their radial growth ($S1_{VMI}$ < $S1_{Con}$) at 5 D, protease activity ($S1_{VMI}$ > $S1_{Con}$) at 3-5 D, amylase activity ($S1_{VMI}$ < $S1_{Con}$) at 3-5 D, and antioxidant levels ($S1_{VMI}$ > $S1_{Con}$) at 3 D. Furthermore, we observed a distinct clustering pattern for gas chromatography-time of flight-mass spectrometry datasets from 5 D extracts of $S1_{VMI}$ and $S1_{Con}$ in principle component analysis (PC1: 30.85%; PC2: 10.31%) and partial least squares discriminant analysis (PLS-DA) (PLS1: 30.77; PLS2: 10.15%). Overall, 43 significantly discriminant metabolites were determined for engendering the metabolic variance based on the PLS-DA model (VIP > 0.7, p < 0.05). In general, a marked disparity in the relative abundance of amino acids ($S1_{VMI}$ > $S1_{Con}$) at 5 D, organic acids ($S1_{VMI}$ > $S1_{Con}$) at 5 D, and kojic acid ($S1_{VMI}$ < $S1_{Con}$) at 5-7 D were observed. Examining the headspace VOCs shared between S1 and S2 in the twin plate for 5 D incubated samples, we observed the relatively higher abundance of C-8 VOCs (1-octen-3-ol, (5Z)-octa-1,5-dien-3-ol, 3-octanone, 1-octen-3-ol acetate) having known semiochemical functions. The present study potentially illuminates the effects of VMI on commercially important A. oryzae's growth and biochemical phenotypes with subtle details of altered metabolomes.

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.

Functional Cosmetic Effect of Porcine Placeta (Porcine Placenta의 기능성 화장품소재 특성)

  • Kim, Bo Young;Kim, Tagon;Kang, Whan Yul;Baek, Hyun;Cheon, Hae Young;Kim, Donguk
    • Korean Chemical Engineering Research
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    • v.48 no.3
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    • pp.327-331
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    • 2010
  • Porcine placenta was treated with alkali, acid and enzyme treatment to obtain extracts. Heavy metal contents such as Pb, As, and Hg were low enough to satisfy cosmetic agent standard. As a result of safety test(MTT assay), porcine placenta extracts showed over 80% of cell viability at $50{\mu}g/ml$, and cell toxicity was relatively lower. From antioxidation test using DPPH free radical scavenging assay, antioxidation effect was highest as 63% at $50{\mu}g/ml$ when porcine placenta was treated with alkali in pH 9. From whitening effect test using tyrosinase inhibition assay, tyrosinase inhibition effect was 30% at $50{\mu}g/ml$ concentration in alkali treated procine placenta, however, the efficiency was lower compared with arbutin or vitamin C. In anti-wrinkle effect test from elastase inhibition assay, elastase inhibition effects were 20~30% at $50{\mu}g/ml$ for 5 kinds of porcine placenta treatments, which was superior to standard, and especially, protease treated extracts showed best results. Skin formulation including 1% porcine placenta was made and the formulation was very stable for temperature and storage period. From this research, porcine placenta extract showed high potential for anti-wrinkle functional cosmetic agent.

Sensory Evaluation and Changes in Physiochemical Properties, and Microflora and Enzyme Activities of Pumpkin-added Kochujang (호박을 첨가하여 제조한 고추장의 숙성 중 성분 변화 및 관능적 특성)

  • Choo, Jong-Jae;Shin, Hyun-Ju
    • Korean Journal of Food Science and Technology
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    • v.32 no.4
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    • pp.851-859
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    • 2000
  • In the present study, changes in physicochemical and sensory properties, and microflora and enzyme activities, by addition of pumpkin (1, 2 and 5% pumpkin on the weight basis), of Sunchang sikhe kochujang, the most famous traditional kochujang, were investigated. Moisture content appeared to be reduced during fermentation, possibly due to exposure to the sun. Patterns of pH changes were rather complicated. pH of control kochujang was gradually reduced from the begining of fermentation up to 60 days of fermentation. Thereafter, it tended to be increased but again reduced after 90 days of fermentation. On the other hand, titratable acidity increased with fermentation reaching at the highest level at 90 day of fermentation and then slowly decreased. The addition of pumpkin induced a reduction in pH and an increase in titratable acidity, but this was due to organic acid contained in pumpkin itself. Contents of reducing sugar and amino nitrogen were increased by the addition of pumpkin in relation to the level of addition. Ethanol content was highly increased by the addition of pumpkin. Bacterial and yeast count, and activities of amylase and protease were not affected by the addition of pumpkin. Sensory evaluation test revealed that the addition of pumpkin improved sweet and savory taste, but color turned to be undesirable. However, none of tested parameters except color showed statistical significance.

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A Study on the Thermal Characterization of Barley ${\beta}-Glucan$ [mixed-linked $(1-3),(1-4)-{\beta}-D-Glucan$] by Differential Scanning Calorimetry (DSC에 의한 보리 ${\beta}-Glucan$ [mixed-linked$(1-3),(1-4)-{\beta}-D-Glucan$의 열적 특성에 관한 연구)

  • Cha, Hee-Sook;Kim, Mi-Ok;Koo, Sung-Ja
    • Korean Journal of Food Science and Technology
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    • v.25 no.1
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    • pp.22-27
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    • 1993
  • Crude ${\beta}-glucan$ extracted from Barley was purified by stepwide enzyme treatment with thermostable ${\alpha}-amylase$, amyloglucosidase and protease. The thermal properties of Barley ${\beta}-glucan$ were investigated by Differential Scanning Calorimetry. Three endotherms have been observed on DSC thermograms of Barley ${\beta}-glucan$. The first endotherm which produced the gelatinization phenomena commonly observed in Barley ${\beta}-glucan$ became the focus of this study. The temperature range and the enthalpy of gelation exhibited maximum values with increasing concentration of Barley ${\beta}-glucan$. Gelating Barley ${\beta}-glucan$ registered an enthalpy of approximately 0.23 cal/g and exhibited onset temperature (To), peak temperature (Tp) and conclusion temperature (Tc) of $48.8^{\circ}C,\;61.2^{\circ}C\;and\;78.5^{\circ}C$ respectively. The temperature and enthalpy of gelatinizing Barley ${\beta}-glucan$ at both alkali and acid conditions were lower than those at pH 7. With salt present, the Tp and Tc of gelating Barley ${\beta}-glucan$ produced lower temperatures than in conditions where salt was absent, and the enthalpy abruptly decreased. However, increasing salt concentrations did not affect the gelation temperature and the enthalpy of Barley ${\beta}-glucan$. The 'true melting' temperature of Barley ${\beta}-glucan$ was near $184^{\circ}C$ and the melting enthalpy was approximately 34.6 cal/g. The Barley ${\beta}-glucan$ decomposition temperature was in the range of $316^{\circ}C{\sim}346^{\circ}C$.

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Effects of Extrusion-Texturization on Defatted Soy Flour for Soy Sauce Fermentation (압출성형기에 의한 전처리가 탈지대두분의 장유 제조특성에 미치는 영향)

  • Oh, Kyeong-Keun;Jung, Heon-Woong;Park, Ji-Yong;Lim, Jae-Kag;Kim, Jae-Cherl
    • Korean Journal of Food Science and Technology
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    • v.25 no.1
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    • pp.1-8
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    • 1993
  • Extruded defatted soy flour (DSF) with twin screw extruder was compared with steam treated DSF for soy sauce fermentation. Independent variables of response surface methodology (RSM) for extrusion were barrel temperature $(145{\sim}165^{\circ}C)$, feed moisture content $(25{\sim}35%)$, and feed rate $(20{\sim}30\;kg/hr)$. Extrusion conditions for production of the extrudates having highest water absorption capacity, lowest bulk density and highest expansion ratio were obtained by regression analysis of each dependent variable. Electron photomicrographs revealed significant increase of porosity inside the koji made from extruded DSF. Therefore, growth of fungi inside the koji made from extrudates of DSF was accelerated to achieve higher activities of protease, ${\alpha}-amylase$, and glucoamylase compared to the koji made from steamed DSF. Although amino acid composition of soy sauces prepared from two different DSF did not show any significant difference, the content of free amino acids in soluble nitrogen compounds was higher $(13%{\pm}2)$ in soy sauce made from extruded SDF. During the whole period of fermentation color of soy sauce made from extrudates of DSF was darker. Sensory evaluation did not reveal any serious off-odor and off-taste in the soy sauce fermented from extruded DSF.

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High-level Expression of Human Procaspase-9 in Escherichia coli and Purification of its GST-tagged Recombinant Protein (대장균을 이용한 세포사멸 유도 단백질 caspase-9의 발현에 관한 연구)

  • Seong, Yeong-Mo;Han, Cheol;Choe, Ju-Yeon;Park, Hyo-Jin;Seong, Geun-Hye;Nam, Min-Gyeong;Kim, Sang-Su;Kim, In-Gyeong;Gang, Seong-Man;Im, Hyang-Suk
    • Korean Journal of Microbiology
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    • v.39 no.4
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    • pp.216-222
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    • 2003
  • Human caspase-9, an essential apoptosis initiator protease, was excessively degraded when expressed in Escherichia coli under the conventional induction condition. To optimize the conditions for induction and develop a rapid purification method for obtaining significant amounts of wild-type procaspase-9, we expressed procaspase-9 as GST fusion in E. coli. The addition of 0.01 mM IPTG as an inducer to the bacterial culture and decreasing the culture temperature to 25oC improved the production of procasapse-9 protein by circumventing proteolytic degradation in E. coli. The wild-type procaspae-9 was purified to approximately 70% purity with relatively high yields using the method developed in this study. In addition, we found that GST-caspase-9 is autocatalytically cleaved after aspartic acid 315, which is the same site for processing in mammalian cells, during expression in E. coli.

Brewing Method and Composition of Traditional Kochuzang (Red Pepper Paste) in Junrabook-do Area (전라북도지방(全羅北道地方) 전통(傳統)고추장의 제법조사(製法調査)와 성분(成分))

  • Cho, Han-Ok;Kim, Jong-Goon;Lee, Hyne-Ja;Kang, Joo-Hoon;Lee, Taik-Soo
    • Applied Biological Chemistry
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    • v.24 no.1
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    • pp.21-28
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    • 1981
  • Brewing method and quality of 15 samples of traditional Kochuzang in Junrabook-do area were investigated. Average combination ratio of glutinous rice: Meju (soybean Koji): red pepper powder was 2 : 0.7 : 1.1. In order to improve the taste of Kochuzang, some amount of natural soysauce was added in the Kochuzang at early stage of fermentation. Average content of sodium chloride was 7.32%; amino nitrogen, 123.3%; total sugar, 31.9%; reducing sugar, 19.4%; ethyl alcohol, 1.13% and the activity of acid protease was 0.07; liquefying amylase, 9.67 and saccharogenic amylase was 15.82. Five samples were selected as excellent Kochuzang through sensory evaluation. These samples were similarly strong in hot and sweet taste with alcoholic flavor, glossy surface and brownish red color.

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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.