• Title/Summary/Keyword: Bacillus protease

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Characteristics of Ju-Back and Effect of Ju-Back Fertilizer on Growth of Crop Plants (주류생산 부산물인 주박의 특성 규명 및 주박이 작물생육에 미치는 영향)

  • Lee, Jung-Hoon;Park, Sung-Min;Park, Chi-Duck;Jung, Hyuck-Jun;Kim, Hyun-Soo;Yu, Tae-Shick
    • Journal of Life Science
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    • v.17 no.11
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    • pp.1562-1570
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    • 2007
  • This experiment was conducted to develop fertilizer which promotes plant growth as well as suppressing pathogenic fungi. The fertilizer was made from the mixture of Ju-Back (Korean rice wine cake) and indigenous rhizosphere-bacterium. The main ingredients of Ju-Back were investigated as 6.04% total nitrogen, 42.59% total carbohydrate, 1.01% available phosphate, 73.42% organic matter, 7.72% potassium oxide, 1.35% calcium oxide, 0.53% magnesium oxide. The enzyme activities of Ju-Back were estimated to be 980 units/g for ${\alpha}-amylase$, 300 units/g for glucoamylase, and 1800 units/g for acid pretense. Indigenous rhizosphere bacteria which produced antifungal agent were isolated from soil, and was selected KMU-13 strain which can antagonize against various plant pathogenic fungi (Botrytis cinerea KACC 40573, Sclerotinia sclerotiorum KACC 41065, Fusairum oxysporum KACC 40052, Pythium aphanidermatum KACC 40156, Phytophthora capsici KACC 40476 and Glomerella cingulata KACC 40299). KMU-13 strain was identified as Bacillus subtilis KMU-13 by biochemical and 16s rDNA analysis. The organic fertilizer was made as prototype which was composed 20% Ju-Back, 70% carrier, 9.7% microorganism cultivated solution, 0.3% trace-element. We also investigated an application of fertilizer using Ju-Back for cultivating lettuce (Lactuca sativar) which were grown in three soil conditions that had chemical fertilizer, barnyard manure, lime power, urea, potassium chloride and superphosphate as a control, the whole quantity (80 kg/10a) of posted fertilizer with the control and the half quantity (40 kg/10a) with the control. The growth characteristics were examined and analysed with several weeks interval from 3 weeks to 8 weeks on head length (cm), head width (cm/head), number of leaf and fresh weight (g/plant). The results are summarized as follows. The head width and fresh weight of lettuce were the highest at posted fertilizer 1 (whole quantity) was applied chemical, organic matter (Ju-Back) and carrier. The head length was the highest at posted fertilizer 2 (whole quantity) was applied Ju-Back only.

Studies on the processing of rapid fermented anchovy prepared with low salt contents by adapted microorganism. -3. Processing of low salt fermented anchovy with proteolytic bacteria and quality stability during storage- (미생물을 이용한 저식염 멸치젓의 속성발효에 관한 연구 -3. 단백질분해세균을 이용한 저식염 멸치젓의 제조 및 저장중의 품질 안정성-)

  • Cha, Yong-Jun;Lee, Kang-Hee;Lee, Eung-Ho;Kim, Jin-Soo;Joo, Dong-Sik
    • Applied Biological Chemistry
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    • v.33 no.4
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    • pp.330-336
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    • 1990
  • In order to Process rapid fermented anchovy with low salt contents, processing condition of rapid fermented anchovy by proteolytic bacteria, and its chemical composition and quality stability during storage were examined. Culture was performed(pH 7.0, $40^{\circ}C$, 45strokes/min) for 15hrs after the addition of 1% of NaCl, 1% of sodium erythorbate and 20m1 of B. licheniformis p-5 cultures($3.2{\times}10^4cells/ml$) to 100g of raw anchovy, and then low salt fermented anchovy as final product was made by adding of several(3% of NaCl, 4% of KCI, 4% of ethyl alcohol(w/v), 0.5% of ginger, 0.5% of garlic powder) for stability and flavor enhancement. During 60days of storage, histamine contents was adequate in a food sanitation aspect, and microflora decreased sharply while volatile basic nitrogen increased slowly. Free amino acids are the major part in unique fermented anchovy taste. The volatile fatty acids is the most important component in the anchovy's flavor. From the results of experiments, it was supposed that rapid fermented anchovy processed with proteolytic bacteria was suitable.

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Raw Starch-digesting Amylase is Comprised of two Distinct Domains of Catalytic and Substrate-Adsorbable Domain: Role of the C- Terminal Region in Raw-Starch-Binding

  • Kim, Cheorl-Ho
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2001.06a
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    • pp.40-45
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    • 2001
  • Raw starch-digesting amylase (BF-2A, M.W. 93, 000 Da) from Bacillus circulans F-2 was converted to two components during digestion with subtilisin. Two components were separated and designated as BF-2A' (63, 000 Da) and BF-2B (30, 000 Da), respectively. BF-2A' exhibited the same hydrolysis curve for soluble starch as the original amylase (BF-2A). Moreover, the catalytic activities of original and modified enzymes were indistinguishable in $K_{m}$, Vmax for, and in their specific activity for soluble starch hydrolysis. However, its adsorbability and digestibility on raw starch was greatly decreased. Furthermore, the enzymatic action pattern on soluble starch was greatly different from that of the BF-2A. A smaller peptide (BF-2B) showed adsorb ability onto raw starch. By these results, it is suggested that the larger peptide (BF-2A') has a region responsible for the expression of the enzyme activity to hydrolyze soluble substrate, and the smaller peptide (BF-2B) plays a role on raw starch adsorption. A similar phenomenon is observed during limited proteinase K, thermolysin, and endopeptidase Glu-C proteolysis of the enzyme. Fragments resulting from proteolysis were characterized by immunoblotting with anti-RSDA. The proteolytic patterns resulting from proteinase K and subtilisin were the same, producing 63- and 30-kDa fragments. Similar patterns were obtained with endopeptidase Glu-C or thermolysin. All proteolytic digests contained a common, major 63-kDa fragment. Inactivation of RSDA activity results from splitting off the C-terminal domain. Hence, it seems probable that the protease sensitive locus is in a hinge region susceptible to cleavage. Extracellular enzymes immunoreactive toward anti-RSDA were detected through whole bacterial cultivation. Proteins of sizes 93-, 75-, 63-, 55-, 38-, and 31-kDa were immunologically identical to RSDA. Of these, the 75-kDa and 63-kDa proteins correspond to the major products of proteolysis with Glu-C and thermolysin. These results postulated that enzyme heterogeneity of the raw starch-hydrolysis system might arise from the endogeneous proteolytic activity of the bacterium. Truncated forms of rsda, in which the gene sequence encoding the conserved domain had been deleted, directed the synthesis of a functional amylase that did not bind to raw starch. This indicates that the conserved region of RSDA constitutes a raw starch-binding domain, which is distinct from the active centre. The possible role of this substrate-binding region is discussed.d.

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Preparation Method of meju by Three Step Fermentation (3단 발효에 의한 메주 제조방법)

  • Kim, Ig-Jo;Lee, Jeong-Ok;Park, Mi-Hwa;Shon, Dong-Hwa;Ha, Yeong-Lae;Ryu, Chung-Ho
    • Korean Journal of Food Science and Technology
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    • v.34 no.3
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    • pp.536-539
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    • 2002
  • This study was investigated for the preparation method of meju by three step fermentation. The steamed soybeans were fermented with nisin-producing Lactococcus lactis subsp. lactis IFO12007 to inhibit the growth of odor-producing Bacilli and promote of normal fermentation of meju. The lactie-fermented soybeans were further fermented with Aspergillus oryzae and Bacillus subtilis. The produced meju by this method showed 54%moisture content and pH 7.0, respectively. The three step fermented meju was soaked in 20% (w/v) brine at $25^{\circ}C$ for 90days. After 30 days, the contents of total free amino acids revealed 4,015 mg% which were higher $3{\sim}5$ times than controls. Among the detected free amino acids, the contents of glutamic acid and leucine showed 925 mg% and 380mg%, respectively. Therefore, the new method of producing meju would be an excellent alternative to improve the quality of soybean fermented food, such as doenjang and kanjang.

Plant Growth Promotion and Biocontrol Potential of Various Phytopathogenic Fungi Using Gut Microbes of Allomyrina dichotoma Larva (장수풍뎅이 유충의 장내 미생물을 이용한 다양한 식물 균류병의 생물적 방제 및 생장촉진)

  • Kim, Joon-Young;Kim, Byung-Sup
    • Research in Plant Disease
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    • v.26 no.4
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    • pp.210-221
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    • 2020
  • This research was executed to select beneficial antagonists from digestive organ of Allomyrina dichotoma larva that can be put on environment friendly control against phytopathogenic fungi. We screened 38 bacterial strains inhibiting mycelial growth against eight plant pathogens through dual culture assay. The 10 strains among 38 bacterial strains were selected as beneficial microbes showing antifungal activity against Botrytis cinerea, Plasmodiophora brassicae, Colletotrichum acutatum and Phytophthora capsici through under greenhouse pot trials. The 10 bacterial strains that shown strongest antifungal activity were classified into 3 genera and 10 species, and identified as the genus Bacillus (DM146, DM152, DH2, and DH16), Paenibacillus (DF30, DH14, and DM142) and Streptomyces (DF137, DM48, and DH92) by morphological characteristics and 16s rRNA gene sequence. The 10 bacterial strains had solubilizing activity of insoluble phosphates, production of IAA (indole-3-acetic acid), β-1,3-glucanase and protease. Among the 10 bacterial strains, DM152 strain was produced significant enhancement of all growth parameters of chili pepper and tomato seedlings under greenhouse condition. Thus, this study demonstrated that gut microbes of Allomyrina dichotoma larva will be useful as a potential biocontrol agent against plant pathogens and biofertilizer.

Fermentation Characteristics and Increased Functionality of Doenjang Prepared with Bamboo Salt (죽염 된장의 발효 특성 및 기능성 증진 효과)

  • Jeong, Min-Woo;Jeong, Ji-Kang;Kim, Sin-Jeong;Park, Kun-Young
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.12
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    • pp.1915-1923
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    • 2013
  • The aim of this study is carried out to evaluate the fermentation characteristics and increased functionality when doenjang was prepared with bamboo salts. Grain type mejus were fermented with mixed starter cultures of Aspergillus oryzae, Bacillus subtilis, and Lactococcus lactis. These mejus were mixed with different kinds of salts-purified salt (PD), sea salt (SD), one-time baked bamboo salt ($1{\times}BD$), three-time baked bamboo salt ($3{\times}BD$), and nine-time baked bamboo salt ($9{\times}BD$)- when doenjangs were prepared. For doenjang fermentation period of 8 weeks at $37^{\circ}C$, the fermentation characteristics of all the groups were compared. The amino type nitrogen content and enzyme activities (protease and ${\alpha}$-amylase) in the samples were significantly increased. In DPPH radical scavenging activities and hydroxyl radical scavenging activities, $9{\times}BD$ (47% and 69%) showed the highest scavenging activities compared to PD (40% and 49%), SD (42% and 57%), $1{\times}BD$ (42% and 64%) and $3{\times}BD$ (45% and 65%) (P<0.05). The anticancer effects of doenjang in HT-29 cancer cells indicated all the groups, especially doenjang prepared with bamboo salts were higher than the others (P<0.05). Apoptosis related genes of Bax and Bcl-2, as well as inflammation related genes of iNOS and COX-2 were regulated by the treatment of doenjangs in HT-29 cancer cells. SD, $1{\times}BD$, $3{\times}BD$, and $9{\times}BD$ increased the expression level of Bax and decreased the expression level of Bcl-2, iNOS, and COX-2. These results suggest that sea salt and bamboo salt especially bamboo salt could improve fermentation characteristics and functionality of doenjang and play an important role in regulating apoptosis and inflammation related genes in cancer cells.

Characteristic Analysis and Production of Short-Ripened Korean Traditional Soy Sauce Added with Rice Bran (미강 첨가량에 따른 단기숙성 간장의 제조 및 특성 분석)

  • Jeong, Su-Ji;Shin, Mee-Jin;Jeong, Seong-Yeop;Yang, Hee-Jong;Jeong, Do-Youn
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.4
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    • pp.550-556
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    • 2014
  • Rice bran contains both excellent nutritional value and functional advantages. Its utilization is limited due to reducing texture and low storage. To satisfy various tastes, Bacillus spp. having high amylase and protease activities were selected. Using the strains, we made whole grain soybean Meju with a reduced manufacturing period by increasing the concentration of total nitrogen. We made soy sauces with mashing ratios of soy bean and rice bran at 10:0, 9:1, 7:3, and 5:5, and then compared their physiochemical properties. After 2 weeks of fermentation, the sugar content increased from 21~22% to 30~32%. However, pH and salinity showed no differences. At a ratio of 9:1, total nitrogen, amino nitrogen content, and total free amino acid contents were the highest at 1.62%, 652.52 mg%, and 8,804.03 mg/kg, respectively, compared to other mashing ratios of soy bean and rice bran. Especially, the contents of aspartic and glutamic acid, which increase delicate flavoring, were higher in our soy sauce compared to those of general traditional soy sauce and brewed soy sauce, which were 504.25 and 1,262.25 mg/kg, respectively. Serine and alanine, which are related to sweet taste, were present at 49.50 and 518.75 mg/kg, respectively, which were the highest among all mixing ratios, at a ratio of 9:1. Compared to general traditional soy sauce and brewed soy sauce, the contents of histamine and tyramine among biogenic amines decreased to 35.85 and 41.04 mg/kg, respectively. Finally, a soy bean and rice bran mixing ratio of 9:1 was determined to be the optimal mixing ratio in the sensory evaluation.

Effect of Urushiol-Free Extracts from Fermented-Rhus verniciflua Stem Bark with Fomitella fraxinea on the Fermentation Characteristics of Doenjang (Soybean Paste) (장수버섯 배양법에 의해 urushiol이 제거된 발효옻 추출물이 된장발효에 미치는 영향)

  • Choi, Han-Seok;Jeong, Seok-Tae;Choi, Ji-Ho;Kang, Ji-Eun;Kim, Eugene;Noh, Jong-Min;Kim, Myung-Kon
    • The Korean Journal of Mycology
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    • v.40 no.4
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    • pp.244-253
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    • 2012
  • The effect of fermented Rhus verniciflua stem bark (FRVSB) extract on the microbial count, enzyme activity, concentrations of free amino acids and organic acids, and physiochemical properties of doenjang (soybean paste) was evaluated during brine fermentation. The FRVSB extract increased the total free amino acid concentration by 1.3-3.1-fold on the $42^{nd}$ day of brine fermentation. After the filtration of brine, the following microbial counts were obtained in the doenjang: bacteria, $0.3{\times}10^8-12.0{\times}10^8$ cfu/g; mold, $3.0{\times}10^4-21.0{\times}10^4$ cfu/g; yeast, $1.0{\times}10^4-2.0{\times}10^4$ cfu/g; Escherichia coli, not detected; and Bacillus cereus, $3.0{\times}10^2-25.0{\times}10^2$ cfu/g. The FRVSB extract addition enhanced the protein and starch degrading activity by 13.8-26.0% and 16.1-35.1%, respectively. The extract increased the total free amino acid content by 1.4-3.0-fold. Lactic acid, acetic acid, and pyroglutamic acid were the predominant organic acids in doenjang. Moreover, the proximate composition, pH, moisture, ash, salt, and amino nitrogen content were increased.