• Title/Summary/Keyword: protease production

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Effect of Bacillus Strains on the Chungkookjang Processing -III. Changes of the Free Amino Acid Contents and Nitrogen Compounds during Chungkookjang Koji Preparation- (균주(菌株)를 달리한 청국장 제조(製造)에 관(關)한 연구(硏究) -제3보(第三報) : 청국장의 유리(遊離) 아미노산(酸) 함량(含量)과 질소성분(窒素成分)-)

  • Suh, Jeong-Sook;Ryu, Myung-Ki;Hur, Yun-Hang
    • Korean Journal of Food Science and Technology
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    • v.15 no.4
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    • pp.385-391
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    • 1983
  • The change of free amino acid contents and nitrogen compounds in the course of the Chungkookjang fermentation that occurred by utilizing Bacillus natto and Bacillus subtilis are to the following effects. pH, during the growth period, that is 6.35 in pH at the first stage of fermentation, were turned into 8.2 after 72 hours. Crude protein content increased irregularly from 16.82%-18% and total sugar decreased. Increasing with the progress of fermentation time, protease activity showed the maximum value between 48-60 hours, but Bacillus natto activated a little than Bacillus subtilis. Amino nitrogen and water soluble nitrogen content increased but difference was found that is, Bacillus natto increased more than Bacillus subtilis. Glutamic acid content was the highest among the contents of free amino acid between both Bacillus sp. and the order of the next contents showed as leucine, phenylalanine, histidine alanine. arginine, but difference was found between Bacillus sp., that is, Bacillus natto was higher than Bacillus subtilis. In view of the results as above, Bacillus natto was excellent than Bacillus subtillus as Bacillus strains of Chungkookjang koji production.

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Characterization of Antibacterial Substance - Producing Bacillus subtilis Isolated from Traditional Doenjang (전통 된장으로부터 분리한 향균물질 생산 Bacillus subtilis의 특성)

  • Ryu, Hyun-Soon;Shon, Mi-Yae;Cho, Soo-Jeong;Park, Seok-Kyu;Lee, Sang-Won
    • Applied Biological Chemistry
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    • v.50 no.2
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    • pp.87-94
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    • 2007
  • A bacterium which has high enzymatic activities such as amylase, cellulase and protease was isolated from Korean traditional soybean food, doenjang. The isolated bacterium was identified to Bacillus subtilis HS25 by the test of morphological and biochemical properties according to Bergey's Manual of Systematic Bacteriology and API 50 CHL kit, and by the 16S rDNA sequence. The isolated B. subtilis HS25 had a potent antibacterial activity against food born causative or pathogenic bacteria. B. subtilis HS25 is endospore forming cell and contained flagella and abundant viscous material at the out layer of cell wall. It was rod type bacterium $(0.5{\sim}0.8{\times}3{\sim}5{\mu}m)$ having biochemical characteristics such as gram staining(+), catalase(+), oxidase(-) and hydrolysis of esculin(+). The optimal medium compositions for production of antibacterial substance in the B. subtilis HS25 were 1% of soluble starch, 0.5% of yeast extract, 0.5% of peptone and 0.05% of MgCl$_2{\cdot}6H_{2}O$. The optimum temperature and pH of the growth of the B. subtilis HS25 was 35$^{\circ}C$ and pH 7.5, respectively. The antibacterial activity was more high in neutral to a little alkaline pH (6.5-10.5) than in acidic pH. The optimal shaking speed to grow and to produce antibacterial substance of the B. subtilis HS25 was 160${\sim}$200 rpm. The optimal culture time for antibacterial activities of the bacterium were shown to be in the range of 12-36 hr.

Assessment of the Quality Characteristics of Mixed-grain Nuruk Made with Different Fungal Strains (곰팡이 균종을 달리하여 제조한 혼합 곡류 누룩의 품질특성)

  • Baek, Seong-Yeol;Kim, Joo-Yeon;Choi, Ji-Ho;Choi, Jeong-Sil;Choi, Han-Seok;Jeong, Seok-Tae;Yeo, Soo-Hwan
    • Journal of the East Asian Society of Dietary Life
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    • v.22 no.1
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    • pp.103-108
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    • 2012
  • Nuruk is a fermented ingredient used for production of traditional Korean rice wine. In this study, quality characteristics of mixed-grain nuruk was made by brewing with different fungal strains was analyzed. Quality elements including enzyme activity and organic acids constituents were measured. The fermentation time of the nuruk did not make a significant difference in terms of its pH, but the acidity and amino acid content for nuruk made from a mixture of two fungal strains was higher than that seen with a single fungal strain. Overall, the enzyme activity for two fungal strain nuruk was higher than that observed for single fungal strain nuruk, with ${\alpha}$-amylase and acidic protease activity in the mixed strain nuruk observed to be more than twice that of the single strain. The major organic acids observed in the manufactured nuruk were identified as acetic, citric, formic, fumaric, lactic, malic and oxalic acids. The total amount of organic acids contained in the nuruk made with the two fungal strain was (2,116.3 mg%). The fungal strains used were A. kawachii SC60 nuruk (1,608.5 mg%) and A. oryzae RIB1353 nuruk (1,146.7 mg%).

Application Potential of Hurdle Technology by Combination of Bacteriocin Produced by Lactobacillus brevis DK25 and Potassium Benzoate (Lactobacillus brevis DK25의 박테리오신과 안식향산칼륨과의 혼용에 의한 Hurdle Technology 적용 가능성)

  • Lim, Sung-Mee
    • Korean Journal of Microbiology
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    • v.47 no.4
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    • pp.364-374
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    • 2011
  • Lactobacillus brevis DK25 isolated from Dongchimi was identified by physiological and biochemical tests and 16S rDNA sequence analysis. Bacteriocin of L. brevis DK25 exhibits inhibitory activity against Enterococcus faecalis and Listeria monocytogenes when using agar well diffusion method. Maximal production of bacteriocin was reached in the beginning of the stationary phase, and inhibitory activity declined after the late stationary phase. This result suggested that bacteriocin was produced in a growth-associated manner. Complete inactivation of bacteriocin activity was observed after treatment with protease, but the activity was stable between pH 4-9 and heat resistant (30 min at $100^{\circ}C$). Bacteriocin showed a concentration-dependent antimicrobial activity against L. monocytogenes KCTC 3569. Moreover, the application experiment showed that combination of bacteriocin (320 AU/ml) with potassium benzoate (0.05%) could significantly reduce the counts of L. monocytogenes KCTC 3569 in mayonnaise during storage at 4 or $25^{\circ}C$ for 10 days. Thus, bacteriocin from L. brevis DK25 may be used for hurdle technology by combination with potassium benzoate in order to increase pathogenic bacteria inactivation in food processing and food safety control.

Fermentation of Black Garlic Wine and its Characteristics (흑마늘 와인의 발효 및 그 특성)

  • Ha, Sung Min;Choi, Hye Jung;Shin, Gyeong Yeon;Ryu, Beung Ho;Joo, Woo Hong
    • Journal of Life Science
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    • v.26 no.7
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    • pp.796-804
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    • 2016
  • In the present study, we screened suitable yeasts for wine fermentation and evaluated the fermentative characteristics of Saccharomyces sp. BCNU 6006 and its anti-oxidant activities. Firstly, various yeasts were isolated from Makgeolli, fruits, and fermented foods. Then, the preliminary selections of suitable yeasts were made using an enzymatic activity assay of glucosidase, glycosidase, protease and tolerance to ethanol and SO2. In addition, the production of biogenic amines and hydrogen sulfide was also monitored. The 9 yeast strains initially selected were determined to belong to the genera Saccharomyces and Kazachtania phylogenetically. We investigated the optimal conditions for wine fermented with black garlic juice (BGJ). The optimal conditions of alcohol fermentation using BGJ were 26 brix, 28℃, and 10 days. Finally, the fermentation products of black garlic wine (BGW) fermented with Saccharomyces sp. BCNU 6006 exhibited 15.03% ethanol, 12 brix of sugar, and pH 4.01. The contents of total polyphenol, total flavonoid, tannin, and 5-HMF compound of BGW were 3.85 mg/ml, 0.51mg/ml, 5.90 mg/ml, and 0.07 mg/ml respectively, lower than that of BGJ. DPPH radical scavenging activity, ABTS radical scavenging activity, and reducing power of BGW were 90.77%, 95.20% and 1.261 respectively, lower than that of BGJ. Superoxide anion (O2-) radical scavenging activity was 94.42%, higher than that of BGJ. Based on the above results, the industrial potential of Saccharomyces sp. BCNU 6006 as a wine-making yeast was confirmed in the present study.

Effect of Red Pepper Varieties on the Microflora, Enzyme Activities and Taste Components of Traditional Kochujang during Fermentation (고추품종을 달리한 전통식 고추장의 숙성중 미생물, 효소활성 및 맛성분의 변화)

  • Shin, Dong-Hwa;Kim, Dong-Han;Choi, Ung;Lim, Mi-Sun;An, Eun-Young
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.6
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    • pp.1050-1057
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    • 1997
  • Microbial counts, enzyme activities and taste components of traditional kochujangs prepared with the powders from 4 different varieties of red pepper, were investigated during 90 days fermentation for the industrial production of traditional kochujang. The viable cell counts of anaerobic bacteria in kochujangs did not change remarkably during fermentation, however, aerobic bacterial counts showed a rapid increase up to 90 days of aging. The yeasts in all kochujang samples increased until 60 days of aging and than decreased. After 90 days of aging, the count of aerobic bacteria in Kumtop kochujang was higher than those of others. The activities of liquefying amylase decreased during the aging, but those of saccharogenic amylase increased at 60 days of aging. The activities of neutral protease were higher than those of acidic protease, and increased during the middle and last period of aging. The major free sugars of kochujang were maltose and glucose, and their contents were higher in Hongkwang kochujang. The major organic acids of kochujang were succinic, formic and citric acid, followed by lactic acid. Succinic acid content in kochujang decreased during fermentation, whereas formic and citirc acids were increased. The major free amino acids were serine, proline, glutamic acid, aspartic acid and alanine. Kumtop kochujang contained the highest amount of total free amino acids. Among the nucleotides and related components in kochujang, cytidine-5-monophosphate was the most abundant component at the begining of aging period, while hypoxanthine increased remarkably during fermentation. Hanwang kochujang was higher in the content of nucleotides than others. Capsaicin contents of kochujang decreased during aging and those of Jangter kochujang was higher than that of others. Sensory evaluation showed that Jangter kochujang was significantly better than Kumtop kochujang in overall acceptability, but there were no appreciable differences in color and flavor.

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β-Xylosidase and β-mannosidase in combination improved growth performance and altered microbial profiles in weanling pigs fed a corn-soybean meal-based diet

  • Liu, Shaoshuai;Ma, Chang;Liu, Ling;Ning, Dong;Liu, Yajing;Dong, Bing
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.11
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    • pp.1734-1744
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    • 2019
  • Objective: In this study, two glycosidases (XMosidases), ${\beta}$-xylosidase and ${\beta}$-mannosidase, were investigated on their in vitro hydrolysis activities of feed and on the improvement of growth performance in vivo in weanling pigs. Methods: Enzyme activities of XMosidases in vitro were evaluated in test tubes and simulation of gastric and small intestinal digestion, respectively, in the presence of NSPase. In vivo study was performed in 108 weaned piglets in a 28-d treatment. Pigs were allotted to one of three dietary treatments with six replicate pens in each treatment. The three treatment groups were as follows: i) Control (basal diet); ii) CE (basal diets+CE); iii) CE-Xmosidases (basal diets+ CE+${\beta}$-xylosidase at 800 U/kg and ${\beta}$-mannosidase at 40 U/kg). CE was complex enzymes (amylase, protease, xylanase, and mannanase). Results: In vitro XMosidases displayed significant activities on hydrolysis of corn and soybean meal in the presence of non-starch polysaccharide degrading enzymes (xylanase and ${\beta}$-mannanase). In vitro simulation of gastric and small intestinal digestion by XMosidases showed XMosidases achieved $67.89%{\pm}0.22%$ of dry matter digestibility and $63.12%{\pm}0.21%$ of energy digestibility at $40^{\circ}C$ for 5 hrs. In weanling pigs, additional XMosidases to CE in feed improved average daily gain, feed conversion rate (p<0.05), and apparent total tract digestibility of crude protein (p = 0.01) and dry matter (p = 0.02). XMosidases also altered the gut bacterial diversity and composition by increasing the proportion of beneficial bacteria. Conclusion: Addition of a complex enzyme supplementation (contained xylanase, ${\beta}$-mannanase, protease and amylase), XMosidases (${\beta}$-xylosidase and ${\beta}$-mannosidase) can further improve the growth performance and nutrient digestion of young pigs.

A Study of the Diversity and Profile for Extracellular Enzyme Production of Aerobically Cultured Bacteria in the Gut of Muraenesox cinereus (갯장어(Muraenesox cinereus) 장으로부터 호기적 조건에서 분리된 미생물의 다양성 및 세포외 효소 생산능 분석에 관한 연구)

  • Lee, Yong-Jik;Oh, Do-Kyoung;Kim, Hye Won;Nam, Gae-Won;Sohn, Jae Hak;Lee, Han-Seung;Shin, Kee-Sun;Lee, Sang-Jae
    • Journal of Life Science
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    • v.29 no.2
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    • pp.248-255
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    • 2019
  • This research confirmed the diversity and characterization of gut microorganisms isolated from the intestinal organs of Muraenesox cinereus, collected on the Samcheonpo Coast and Seocheon Coast in South Korea. To isolate strains, Marine agar medium was basically used and cultivated at $37^{\circ}C$ and pH7 for several days aerobically. After single colony isolation, totally 49 pure single-colonies were isolated and phylogenetic analysis was carried out based on the result of 16S rRNA gene DNA sequencing, indicating that isolated strains were divided into 3 phyla, 13 families, 15 genera, 34 species and 49 strains. Proteobacteria phylum, the main phyletic group, comprised 83.7% with 8 families, 8 genera and 26 species of Aeromonadaceae, Pseudoalteromonadaceae, Shewanellaceae, Enterobacteriaceae, Morganellaceae, Moraxellaceae, Pseudomonadaceae, and Vibrionaceae. To confirm whether isolated strain can produce industrially useful enzyme or not, amylase, lipase, and protease enzyme assays were performed individually, showing that 39 strains possessed at least one enzyme activity. Especially the Aeromonas sp. strains showed all enzyme activity tested. This result indicated that isolated strains have shown the possibility of the industrial application. Therefore, this study has contributed for securing domestic genetic resources and the expansion of scientific knowledge of the gut microbial community in Muraenesox cinereus of South Korea.

Studies on the Brewing of Kochuzang (Red Pepper Paste) by the Addition of Yeasts (효모첨가(酵母添加)에 의(依)한 고추장의 양조(釀造)에 관(關)한 연구(硏究))

  • Lee, Taik-Soo
    • Applied Biological Chemistry
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    • v.22 no.2
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    • pp.65-90
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    • 1979
  • This study was conducted to establish the brewing method which would be useful for the production of Kochuzang. Kojis, which were made from various materials and microorganisms under a covered condition, were investigated and compared. Yeasts (Saccharomyces rouxii and Torulopsis versatilis) were added to Kochuzang, and the enzyme activity, microflora, chemical composition, nitrogen content, alcohol content and free sugars of Kochuzang were investigated and analysed. The results obtained are as follows: 1. Koji making (1) Glutinous rice-soybean group was superior to glutinous rice group in the saccharogenic and liquefying amylase activities of three day-Koji. (2) Protease activity (acid, neutral and alkaline) of glutinous rice-soybean Koji, which was inoculated with Aspergillus oryzae A, was increased till the 5th day, while other groups showed maximum activity after the 3rd day. (3) The maximum cellulose activity of Aspergillus oryzae B-Koji and A-Koji was observed after the 2nd day and the 3rd day, respectively. High cellulose activity of Aspergillus oryzae B-Koji and A-Koji was respectively shown in glutinous rice group and glutinous rice-soybean group at maximum. (4) Compared with glutinous rice Koji, glutinous rice-soybean Koji gave larger number of yeast and aerobic bacteria. 2. Kochuzang Fermentation (1) Each Kochuzang group shoved different liquefying and saccharogenic amylase activities. The highest activities were generally shown in 10 to 40 days after mashing and remarkably reduced in the last stage of aging. (2) Protease activities of each group were strong in order of acid, neutral and alkaline protease. Especially acid protease showed highest activity at the 40th to 50th day Kochuzang. (3) Each group showed maximum cellulase activity in the 40th and 50th day-Kochuzang and then decreased. (4) Osmophilic yeast of yeast-added Kochuzang after one-month aging was distinctively outnumbered compared with non-yeast-added Kochuzang, but two groups were similar after two months. (5) Yeast-added group and non-added group gave almost the same number of halophilic lactic acid bacteria in Kochuzang, but the non-added group gave slightly larger number of aerobic bacteria than the yeast-added group. (6) Amino nitrogen contents in all test group were increased rapidly till the 60th day of Kochuzang aged. After that the contents were increased slowly. (7) Ethyl alcohol contents of 20day-fermented Kochuzang were high in order of Saccharomyces rouxii-added group, Torulopsis versatilis-added group, Saccharomyces rouxii and Torulopsis versatilis mixed group and non-yeast-added group. But all test group showed about 2% in ethyl alcohol content after 40days of aging. (8) Alcohol content in the 7 month-aged Kochuzang of all test groups was high in order of ethyl alcohol, n-butyl alcohol, n-propyl alcohol and iso-propyl alcohol. Torulopsis versatilis-added group had the highest value of ethyl alcohol, n-propyl alcohol and n-butyl alcohol. (9) Reducing sugar in Kochuzang was increased after 20 days of aging compared with the 10days-ferment. The reducing sugar content in Saccharomyces rouxii-added group was distinctively small compared with that of other groups, decreasing after 30days of aging. (10) Rhamnose, fructose, glucose and maltose were isolated from the 10 day fermented Kochuzang. Raffinose was also found after 300 days-aged group, and fructose content was high in the 300days-aged Kochuzang. However, glucose content was smaller than that of 10days-fermented Kochuzang. (11) For the organoleptic tests of Kochuzang, taste, flavour and color of yeast-added group were superior to the non yeast-added group. Especially the complex yeast group among the yeast added groups were the best of all. Yeast-added group after 300 days of aging took higher paint in flavour test than that of non-added group. Therefore, brewing method like complex yeast added group seems to be advantageous for short time brewing Kochuzang.

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Properties of Plywood Bonded with Adhesive Resins Formulated with Enzymatically-Hydrolyzed Rapeseed Flour (유채박의 효소 가수분해물로 조제한 접착제를 사용한 합판의 접착특성)

  • Yang, In;Han, Gyu-Seong;Choi, In-Gyu;Kim, Yong-Hyun;Ahn, Sye-Hee;Oh, Sei-Chang
    • Journal of the Korean Wood Science and Technology
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    • v.40 no.3
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    • pp.164-176
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    • 2012
  • In the present study, rapeseed flour (RSF), which is a by-product from the production of edible oil and biodiesel extracted from rapeseed, was used to develop alternative adhesives for the production of plywood panels. To examine the effects of the enzyme on the adhesive properties and formaldehyde emission of the RSF-based adhesive resins, three enzymes, such as cellulase (CEL), pectinase (PEC) and protease (ALC), were used either separately or together. As a crosslinking agent, PF prepolymers, which were prepared with 1.5, 1.8 and 2.1 mole formaldehyde and 1 mol phenol (1.8-, 2.1- and 2.4-PF), were added into the RSF hydrolyzates. The adhesive resins formulated with CEL- or CEL-PEC-RSF hydrolyzates and 1.8-F/P PF prepolymers exhibited excellent adhesive strengths and formaldehyde emission. The tensile shear strength and formaldehyde emission of the plywood panels bonded with the formulate resins were satisfied with the minimum requirement of the KS standard for ordinary plywood panels (0.6 N/$mm^2$). In addition, formaldehyde emissions of the plywood panels approached to that of E0 specified in the KS standard (0.5 mg/${\ell}$), and even had much better than those of commercial UF glue mixes. Overall, the use of RSF-based adhesive resins for the production of plywood panels might provide durable adhesive properties and an environmentally friendly substitute for petroleum-based adhesive resins. However, further researches - the increase of solid content of RSF-based adhesives for reducing press time and the microscopic observation of plywood specimen for identifying the relationship between tensile shear strength and the penetration of adhesives into wood structure - are required to commercialize the RSF-based adhesives.