• Title/Summary/Keyword: Acid protease

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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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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.

Effects of Capsosiphon fulvescens Extracts on Essential Amino Acids Absorption in Rats (매생이 열수추출물이 흰쥐의 필수아미노산 흡수에 미치는 영향)

  • Kim, Hyo-Young;Kim, In-Hye;Nam, Taek-Jeong
    • Journal of Life Science
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    • v.19 no.11
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    • pp.1591-1597
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    • 2009
  • The aim of this study was to examine the effects of Capsosiphon fulvescens extract (CFE) on levels of essential amino acids in serum and the rate of protein digestion. CFE contains approximately 68% carbohydrate as fiber, 17% crude ash, and 14% crude protein. Experimental rats were divided two groups, one group receiving a normal diet and the other a normal diet containing 5% CFE. To analyze the absorption of essential amino acids into the hepatic portal vein, blood from the hepatic portal vein was collected between 10 a.m. and 5 p.m. for 10 days. In vitro protein digestion was decreased in the group fed 5% CFE; these data indicated that CFE affected protease activity. We measured the absorption of essential amino acids in the serum of the hepatic portal vein, at 30-, 60-, 90-, and 120 minutes after feeding. Although there was no difference in the concentration of total essential amino acids between the two groups, the 5% CFE-fed animals had a decreased rate of absorption. Absorption of Lys and Thr into the hepatic portal vein was lower in the CFE-fed group than in the control group. The rate of absorption of Met was delayed nearly 50% in the CFE-fed group compared to the control group. On the other hand, the rate of absorption of Leu, Ile, and Val was increased; Phe showed the same. Therefore, we suggest that CFE could affect protein metabolism by increasing or decreasing the absorption rate of essential amino acids.

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.

Fermentation Characteristics of Kochujang Prepared with Various Salts (소금의 종류를 달리한 고추장의 발효 특성)

  • Kim, Dong-Han;Yang, Sung-Eun;Rhim, Jong-Whan
    • Korean Journal of Food Science and Technology
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    • v.35 no.4
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    • pp.671-679
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    • 2003
  • Effects of sun-dried, refined, seaweed, and bamboo salts on the quality of kochujang were studied by measuring enzymatic, microbial, and physicochemical characteristics of kochujang during 12 weeks of fermentation. Yeast count was low in the bamboo-salt kochujang, whereas that of aerobic bacteria was low in the seaweed-salt kochujang. Acid protease activity was high in the bamboo-salt kochujang, whereas amylase activity did not show any remarkable difference. Color change was lowest in the bamboo-salt kochujang. Water activities of all kochujangs decreased during fermentation with the lowest shown in the refined-salt kochujang. Consistency of seaweed-salt kochujang was the highest. Total sugar content was higher, whereas ORP was lower in the seaweed- and bamboo-salt kochujangs. Titratable acidity changed slightly in the bamboo-salt kochujang. Reducing sugar content was the lowest, whereas ethanol content was the highest in the refined-salt kochujang. Amino and ammonia nitrogen contents of kochujangs increased up to the middle of fermentation period then decreased with lower changes observed in the sun-dried and refined-salt kochujangs. Results of sensory evaluation indicated that the bamboo-salt kochujang was more acceptable than the sun-dried salt kochujang in taste, color, and overall acceptabilities.

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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Isoflavone Contents, Antioxidative and Fibrinolytic Activities of Some Commercial Cooking-with-Rice Soybeans (시판되는 밥밑콩류의 이소플라본 함량, 항산화활성 및 혈전용해활성)

  • Oh, Hae-Sook;Park, Young-Hoon;Kim, Jun-Ho
    • Korean Journal of Food Science and Technology
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    • v.34 no.3
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    • pp.498-504
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    • 2002
  • Isoflavone (daidzein and genistein) contents, and antioxidative and fibrinolytic activities of seven commercial Korean cooking-with-rice soybeans, including Seonbikong, Chungtae, Gangnamkong, Whangtae, Geodoo, Seolitae, and Wooltalikong were investigated. Daidzein and genistein were not found in Selitae, nor was didzein in Gangnamkong and Geodoo. Total daidzein and genistein levels in Chungtae and Whangtae were 500 and 1550 mg per kg, respectively. Wooltalikong, Whangtae, and Gangnamkong had very high electron donation abilities, over 90%, but Seonbikong and Chungtae showed significantly lower activities. SOD-like activities were also the highest in Gangnamkong and Wooltalikong. Fibrinolytic activities in Seonbikong, Whangtae, and Gangnamkong were similarly strong. Fibrinolytic substances purified from protease inhibitors or activated under various pH or heat treatment conditions, were different among the soybean varieties. This study revealed that, although several cooking-with-rice soybeans were poor in isoflavones, Wooltalikong and Gangnamkong could be good sources for functional products due to their strong antioxidative activities, and heat- and acid-resistant proteolytic abilities.

Optimization of an Industrial Medium and Culture Conditions for Probiotic Weissella cibaria JW15 Biomass Using the Plackett-Burman Design and Response Surface Methodology

  • Yu, Hyung-Seok;Lee, Na-Kyoung;Kim, Won-Ju;Lee, Do-Un;Kim, Jong-Ha;Paik, Hyun-Dong
    • Journal of Microbiology and Biotechnology
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    • v.32 no.5
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    • pp.630-637
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    • 2022
  • The objective of this study was to optimize industrial-grade media for improving the biomass production of Weissella cibaria JW15 (JW15) using a statistical approach. Eleven variables comprising three carbon sources (glucose, fructose, and sucrose), three nitrogen sources (protease peptone, yeast extract, and soy peptone), and five mineral sources (K2HPO4, potassium citrate, ⳑ-cysteine phosphate, MgSO4, and MnSO4) were screened by using the Plackett-Burman design. Consequently, glucose, sucrose, and soy peptone were used as significant variables in response surface methodology (RSM). The composition of the optimal medium (OM) was 22.35 g/l glucose, 15.57 g/l sucrose, and 10.05 g/l soy peptone, 2.0 g/l K2HPO4, 5.0 g/l sodium acetate, 0.1 g/l MgSO4·7H2O, 0.05 g/l MnSO4·H2O, and 1.0 g/l Tween 80. The OM significantly improved the biomass production of JW15 over an established commercial medium (MRS). After fermenting OM, the dry cell weight of JW15 was 4.89 g/l, which was comparable to the predicted value (4.77 g/l), and 1.67 times higher than that of the MRS medium (3.02 g/l). Correspondingly, JW15 showed a rapid and increased production of lactic and acetic acid in the OM. To perform a scale-up validation, batch fermentation was executed in a 5-l bioreactor at 37℃ with or without a pH control at 6.0 ± 0.1. The biomass production of JW15 significantly improved (1.98 times higher) under the pH control, and the cost of OM was reduced by two-thirds compared to that in the MRS medium. In conclusion, OM may be utilized for mass producing JW15 for industrial use.

가금에서 분리된 유산균의 생리적 특성 및 급여효과

  • 김상호
    • Proceedings of the Korea Society of Poultry Science Conference
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    • 2002.11a
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    • pp.64-84
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    • 2002
  • These studies were conducted to evaluate the Properties of lactic acid Producing bacteria(LAB), isolated from broiler and laying hens cecum and select the optimum strains to improve the performance, environment of poultry house, immunity, and intestinal microflora of broiler and laying hens. In experiment I , 23 LAB strains were isolated from broiler and laying hens cecum as a colony form. Six strains were selected by acid tolerance, bile salt tolerance, viability, enzyme release, antagonism, and antibiotics susceptibility. In Experiment II, selected LABs from Ex. 1 were conducted to investigate the effects of feeding various Lactobacillus on performance, nutrients digestibility, intestinal microflora, villi development and observation of epithelium surface, blood chemicals and fecal noxious gas of broiler chicks. One thousand eighty one day old broiler chicks were fed into Lactobacillus crispatus avibrol(LCB), Lactobacillus reuteri avibro2(LRB), Lactobacillus crispatus avihen1(LCH), and Lactobacillus vaginalis avihen2(LVH) at the level of 10$^4$ and 10$\^$7/cfu/g diet. Weight gam of chicks fed Lactobacillus tended to increase from the first week and was higher from 50 to 100g in Lactobacillus treatments than control. Feed intake and feed conversion were not statistically different of all treatments. Dry Matter digestibility of Lactobacillus treatments was prone to improve compared to that of control, but was not significantly different. Protein and Ca digestibility were also tended to improve in Lactobacillus treatments relative that of control. Lactobacillus treatments showed improved tendency in crude ash and fat compared to those of control, whereas phosphorus digestibility was not consistency. Nutrients digestibilities of bird fed LCH were superior to those of other treatments, It showed significantly higher in Ca and P digestibility than control(P〈0.05). Total Lactobacillus spp. of birds fed various Lactobacillus was significantly higher in illeum for five weeks(P〈0.05), but was not different at cecum. Yeast was thought to be not completely attached to intestinal lumen for one week. However, total number of yeast was significantly increased in cecum and illeum of three weeks old chicks (P〈0.05). The number of anaerobes exhibited to tendency the increase in Lactobacillus treatments from one week old of age at both ileum and cecum.

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Studies on Rheological Characterization of Barley ${\beta}-Glucan$ [mixed-linked $(1-3),(1-4)-{\beta}-D-Glucan$] (보리 ${\beta}-Glucan$ [mixed-linked $(1-3),(1-4)-{\beta}-D-Glucan$의 리올로지 특성)

  • Kim, Mi-Ok;Cha, Hee-Sook;Koo, Sung-Ja
    • Korean Journal of Food Science and Technology
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    • v.25 no.1
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    • pp.15-21
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    • 1993
  • Crude ${\beta}-glucan$ extracted from Barley was purified by stepwise enzyme treatment (Thermostable ${\alpha}-amylase$, amyloglucosidase, protease). The Intrinsic Viscosity $[{\eta}]$ of the purified ${\beta}-glucan$ was determined by Cannon Fenske Capillary Viscometer (size 50, Cannon Instruments, State, College pa.) at different pH (2, 4, 7, 9, 11) and various salt concentration (0.01 M, 0.03 M, 0.05 M, 0.07 M, 0.1 M and 0.2 M). The $[{\eta}]$ of purified ${\beta}-glucan$ was ranged from $0.997{\sim}2.290\;dl/g$. The $[{\eta}]$ of purified ${\beta}-glucan$ at both alkali, acid condition were lower than those at pH 7. However, the alkali condition of puified ${\beta}-glucan$ solution showed less $[{\eta}]$ than the acid condition of this solution. From 0 M to 0.2 M salt concentration, the $[{\eta}]$ of purified ${\beta}-glucan$ solution was decreased to 0.03 M then increased to 0.05 M NaCl and remained constant to 0.2 M NaCl. The chain stiffness parameter of purified ${\beta}-glucan$ was not affected by temperature from $15^{\circ}C$ to $65^{\circ}C$. The shear rates of various ${\beta}-glucan$ conditions were determined by Bohlin Rheometer (Lund, Sweden). The ${\beta}-glucan$ concentration of 1.0 g/dl and 2.0 g/dl behaved as Newtonian fluid. However, above the concentration of 3.0 g/dl ${\beta}-glucan$ solution, it showed thixotropic and psedoplastic characteristics. Barley ${\beta}-glucan$ appears a damping at 0.5 frequency for the 4.0 g/dl solution. Below 0.5 frequency, it appears a viscous behavior property and above 0.5 frequency, it appears a elastic behavior property.

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