• Title/Summary/Keyword: Acid protease

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Isolation and characterization of a noval membrane-bound cytochrome $C_{553}$ from the strictly anaerobic phototroph, heliobacillus mobilis

  • Lee, Woo-Yiel;Bla;Kim, Seung-Ho
    • Journal of Microbiology
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    • v.35 no.3
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    • pp.206-212
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    • 1997
  • Heliobacillus mobilis is a strictly anaerobic Gram-positive bacterium which contains a primitive Photosystem I-type reaction center. The membrane-bound cytochrome $C_{553}$ from the heliobacterium suggested to be the immediate electron donor to the photooxidized pigment (P798+) has been isolated and characterized. The heme protein was visualized as a major component with an apparent molecular size of 17kDa in TMBZ-staining analysis of the membrane preparation and showed characteristic $\alpha$ (552.5 nm), $\beta$ (522nm), and Soret absorption (416 nm) peaks of a typical reduced c-type cytochrome in the partially purified sample. The internal 43 amino acid sequence of the electron donor was obtained by chemical agent and protease treatments followed by N-terminal sequencing of the resulting fragments. The internal sequence carries lots of lysine residues and a Cys-X-X-Cys-His sequence motif which are the characteristics of typical c-type cytochromes. The analysis of the sequence by FAST or FASTA program, however, did not show any significant similarity to other known heme proteins.

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Characterization of Bacteriocin Production by Lactococcus lactis LAB3113 Isolated from Kimchi

  • Shin, Jong-Yeun;Cheol Ahn
    • Preventive Nutrition and Food Science
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    • v.2 no.2
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    • pp.101-108
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    • 1997
  • A lactic acid bacterium LAB3113, isolated from traditionally fermented Kimchi was found to produce bacteriocin whose activity was very specific toward lactobacilli and not effective against any other bacteria. Lactobacilli affected by the inhibitory substance included Lactobacillus delbrueckii-lactis, L. johnsonii, L. gsseri, and L. curvatus. Based upon biochemical and physiological characteristics, LAB3113 was classified as Lactococcus lactis, and its bacteriocin was named as lactococcin K3113. Lactococcus lactis. LAB3113 produced bacteriocin at th early stage of growth and the concentration of the bacteriocin did not decrease even after alt stationalry phase. Optimal temperature of bacteriocin production was $25^{\circ}C$ at the initial pH 7.0. Partially purified lactococcin K3113 was completely inactivated by protease, but not affected by lipase, lysozyme and RNase. The bacteriocin was very heat-stable even after autoclaving for 20 min. It was also stable in pH changes, an was not affected by th presence of solvents. lacotococcin K3113 appeared to act in bactericidal mode against L. delbrueckii-lactis ATCC4797. Molecular weight of lactococcin K3113 was calibrated as 10,500 dal by SDS-PAGE an activity staining. Lactococcus lactis LAB3113 had four residential plasmids of 3.7kb, 11.2kb, 15.5kb, and 48kh in molecular sizes. Plasmid profile analysis of mutant strain revealed that 15.5 kb plasmid was re-sponsible for the production of lactococcin K3113 and its immunity to the bacteriocin.

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Isolation of a Lactococcus lactis Strain Producing Anti-staphylococcal Bacteriocin

  • Yang, Jung-Mo;Moon, Gi-Seong
    • Food Science of Animal Resources
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    • v.38 no.6
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    • pp.1315-1321
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    • 2018
  • Bacteriocin is ribosomally synthesized by bacteria and inhibits closely related species. In this study we aimed at isolating lactic acid bacteria producing bacteriocin presenting anti-staphylococcal activity. A Lactococcus lactis strain was isolated from kimchi for the purpose and identified by 16S rRNA gene sequencing. As preliminary tests, optimal culture conditions, stabilities against heat, solvents, and enzymes treatments, and type of action (bacteriostatic or bactericidal) of the bacteriocin were investigated. The optimal culture conditions for production of the bacteriocin were MRS broth medium and $25^{\circ}C$ and $30^{\circ}C$ culture temperatures. The bacteriocin was acidic and the activity was abolished by a protease treatment. Its stability was maintained at $100^{\circ}C$ for 15 min and under treatments of various organic solvents such as methanol, ethanol, acetone, acetonitrile, and chloroform. Finally, the bacteriocin showed bactericidal action against Staphylococcus aureus where 200 AU/mL of the bacteriocin decreased the viable cell count (CFU/mL) of S. aureus by 2.5 log scale, compared with a control (no bacteriocin added) after 4-h incubation.

Studies on the Chemical Compositions and Quality of Red Pepper Paste Brewed with Different Raw Materials (담금원료(原料)에 따른 고추장의 성분(成分)과 품질(品質)에 관(關)한 연구(硏究))

  • Park, Soo-Woong;Park, Yoon-Joong
    • Korean Journal of Agricultural Science
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    • v.6 no.2
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    • pp.205-212
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    • 1979
  • Red pepper paste were prepared by using various raw materials such as rice, glutinous rice, polished barley, polished wheat and corn powder, and their chemical compositions including reducing sugar, amino-N and ethanol were analyzed during the period of aging in order to elucidate effects of these starchy materials on the compositions and qualities of the products. The results obtained are summarized as follows: 1) Koji of these materials were manufactured respectively by inoculating a strain of Asporyzae and their activities of protease and saccharifying amylase were determined. Wheat koji was found to have the highest level of protease activity among the koji, while glutinous rice koji had the strongest amylase activity. 2) Contents of moisture, crude protein, crude fat, and sodium chloride in the red pepper pastes were not changed significantly, however total sugar content was decreased during the period of aging. 3) After 60 days of aging, the highest amount (160mg%) amino nitrogen was detected in the red pepper paste of polished wheat, but higher levels of reducing sugar and ethanol contents were detected in that of glutinous rice. 4) Amino acids in the products were analyzed after 60 days of aging: a) Total 17 amino acids were detected. b) Free amino acid ratio to total amino acid content was approximately 35 percent in average. c) Free glutamic acid was higher than any other free amino acid In the amount and free ratio, but free methionine was lower. d) Lysine content was relatively high, while histidine and glycine were detected in trace. 5) Sensory tests including color, odor and taste on the products resulted in the highest mark for the red pepper paste of glutinous rice but in the lowest mark for that of corn powder.

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Studies on the N-Compounds during Chung-Kook-Jang Meju Fermentation -Amino acids of oligopeptides formed during Chung-Kook-Jang Fermentation- (청국장(淸國醬)메주 발효과정중(醱酵過程中)의 질소화합물(望素化合物)의 소장(消長)에 관(關)한 연구(硏究)(II) -저급(低級) peptide와 구성(構成)아미노산(酸)에 관(關)하여-)

  • Park, Ke-In
    • Applied Biological Chemistry
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    • v.15 no.2
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    • pp.111-142
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    • 1972
  • An experimental Chung-Kook-Jang was prepared using the strain Bacillus subtilis sp. isolated by the author Samples were taken in 12 hrs interval during the fermentation and the oligopeptides were separated by the method of molecular sieving using the ion exchange resin column of Dowex-50. Only the X-16 fraction of oligopeptides was taken and the components of oligopeptides were developed in two dimensional thin layer chromatograms. The each peptide spot was eluted and each peptide was isolated. The pattern and kinds of amino acids, and N and C-terminal amino acids were studied. Fourteen different oligopeptides could be detected by the two dimensional thin layer chromatography, all of which were consisted of $4{\sim}9$ kinds of amino acids. No dipeptides and no tripeptides could be found. The N and C-terminal amino acids and the residual component amino acids of all these 14 peptides could be summarized as the follows. [P]-I. Pro (Cys Ala Asp Trp Ile Val) Glu [P]-II. Val (His Arg Glu Thr Ala Met) Asp [P]-III. Glu (Cys Lys Asp Thr Met) Ala [P]-IV. Glu(His Ser Ala) Met) [P]-V. Ile (Cys Asp Arg Gly Pro T.p Phe) His [P]-VI. Gly(Asp ser) Lys [P]-VII. Thr(Pro Tyr Phe) Asp [P]-VIII. Phe(Tyr Leu Ile) Val [P]-IX. Trp (Phelle) Thr [P]-X. Ile (Arg Leu) Phe [P]-XI. Asp (Lys His Ser Gly Glu Pro) Ala [P]-XII. Glu (Cys Asp Gly) Ser [P]-XIII. Ala (Arg Tyr) Glu [P]-XIV. Met (Glu Ala) His It appears that the protease of the Bacillus subtilis K-27 syrain has rather wider range of specificity than proteases of Aspergoillus soya, pepsin, chymotrypsin, and trypsin.

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Bactericidal Effect of Bacteriocin of Lactobacillus plantarum K11 Isolated from Dongchimi on Escherichia coli O157

  • Lim, Sung-Mee;Im, Dong-Soon
    • Journal of Food Hygiene and Safety
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    • v.22 no.3
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    • pp.151-158
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    • 2007
  • Among 68 strains of lactic acid bacteria (LAB) isolated from Dongchimi, a strain K11 was selected due to its bactericidal activity against Escherichia coli O157 The strain K11 was identified as Lactobacillus plantarum, based on physiological and biochemical characteristics. In the late exponential phase, La. plantarum K11 showed maximum bacteriocin activity (12,800 BU/mL) and maintained until the early stationary phase. The bacteriocin activity was completely inactivated by all the proteolytic enzymes such as pepsin, protease, proteinase K, papain, chymotrypsin, and trypsin, but the activity was not affected by catalase, a-amylase, lysozyme, and lipase, suggesting proteinaceous nature of the bacteriocin. Additionally, this activity was not affected in the pH range from 3.0 to 9.0 and under storage conditions like 30 days at -20,4, or $25^{\circ}C$. Although the bacteriocin activity was absolutely lost after 15 min treatment at 121, it was relatively stable at $70^{\circ}C$ for 60 min or $100^{\circ}C$ for 30 min. The activity was disappeared by treatment with acetone, benzene, ethanol, or methanol, but it was not affected by treatment with chloroform or hexane. The antibacterial activity of the bacteriocin was good against some LAB including Lactobacillus spp., Enterococcus spp., and Streptococcus spp., but not against food-borne pathogens such as Bacillus spp., Listeria spp., and Staphylococcus spp. as well as yeasts and molds. Especially, some intestinal bacteria such as Enterobacter aerogenes and E. coli were significantly affected by the bacteriocin of La, plantarum K11. Furthermore, the addition of 640 BU/mL resulted in the complete clearance of E. coli O157 after 10 hr.

Effects of Anti-Microbial Materials on Storages of Low Salted Doenjang (항균물질을 첨가한 저식염 된장의 저장성)

  • Kim, Jeong-Rye;Kim, Yon-Kyung;Kim, Dong-Han
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.11
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    • pp.1864-1871
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    • 2013
  • The effect of additives on the quality of low salted doenjang was investigated during storage. Amylase activity gradually decreased during storage and protease activity decreased after four weeks. The number of yeast was lower in the mustard or ethanol added groups without a difference in bacterial count. The L- and b-values decreased gradually during storage with lower total color difference (${\Delta}E$) in garlic added doenjang. Gas production was reduced in the ethanol or mustard added groups. Titratable acidity and acid values were low in the ethanol and ethanol-garlic added ones. A reducing sugar content was higher in the groups with added additives. Ethanol decreased to the largest extent in mustard added doenjang. Amino-type nitrogen decreased in ethanol added doenjang, whereas ammonia-type nitrogen was low in the ethanol or mustard added groups. The taste, flavor, and overall acceptability of doenjang were significantly higher in the ethanol or garlic added groups than in the other groups.

Changes in Microflora, Enzyme Activities and Microscopic Structure of Waxy Rice and Steeping Water in Response to Different Steeping Conditions During Preparation of Gangjung (강정 제조시 찹쌀의 수침조건에 따른 찹쌀과 수침액의 미생물상, 효소활성 및 미세구조 변화)

  • Kim, Haeng-Ran;Kim, Kyung-Mi;Kim, Kwang-Ok
    • Korean Journal of Food Science and Technology
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    • v.41 no.6
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    • pp.644-651
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    • 2009
  • This study was conducted to investigate the effects of steeping periods and temperatures of waxy rice on the microflora and enzyme activities of steeping water and waxy rice, as well as on the microscopic structures of waxy rice during the preparation of gangjung. When the steeping water was 15 and $35^{\circ}C$, yeast and lactic acid bacteria were the major microflora after 1 day. After 11 days, Lactobacillus spp. were the major bacteria, while Bacillus spp. were the primary microflora after 21 days. At $25^{\circ}C$ the primary species were Bacillus spp., Leuconostoc spp., and Bacillus spp. after 1, 11 and 21 days, respectively. The $\alpha$-amylase activity of steeping water increased proportionally with the increase of temperature and period, while the protease activity tended to increase for 11 days, but decreased at 21 days. The enzyme activities of the steeped waxy rice, especially the $\alpha$-amylase activity, decreased with higher steeping temperature, while the $\beta$-amylase and glucoamylase activity remained constant for up to 21 days. Observation using scanning electron microscopy indicated that intracellular cleavage of steeped waxy rice endosperm resulted in a rough surface morphology due to the exposure of polygonal starch granules. Tiny pinholes on the surface of waxy rice starch granules were also observed after steeping for 11 days.

The Identification and Physiological Properties of Lactobacillus plantarum JK-01 Isolated from Kimchi (김치로부터 분리한 Lactobacillus plantarum JK-01의 동정 및 생리적 특성)

  • Cho, Jin-Koo;Li, Guan-Hao;Cho, Sung-Jin;Yoon, Yoh-Chang;Hwang, Seong-Gu;Heo, Kang-Chil;Choe, Il-Shin
    • Food Science of Animal Resources
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    • v.27 no.3
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    • pp.363-370
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    • 2007
  • In order to identify probiotic microorganisms, 25 isolates of Lactobacillus sp. were selected from kimchi based on their growth rates, lactic acid production and salt tolerance. The isolate JK-01 was identified as Lactobacillus plantarum by the API kit and 16S rDNA analysis (99.9% of homology), and named as L. plantarum JK-01. The maximum number of L. plantarum JK-01 was reached at 18 hr fermentation in MRS broth and the pH gradually decreased to 4.5. L. plantarum JK-01 showed high enzyme activities for xylanase, amylase, protease, and phytase on MRS agar plates containing each substrate. L. plantarum JK-01 showed high resistance to acidic pH and bile salts, and grew well even at pH 2.0 and 1.0% bile salt. In particular, L. plantarum JK-01 showed high heat stability as shown by $3.3{\times}10^3$ CFU/mL at $60^{\circ}C$. The isolate showed remarkable antimicrobial activity against E. coli in MRS broth based on its disappearance after 18 hr and clear zone formation using a paper disk assay. These results suggest that L. plantarum JK-01 may be probiotic in nature.

Rapid Fermentation of Fish Sauce and Its Kinetics (어장유의 속성발효와 동력학적 고찰)

  • KIM Byeong-Sam;PARK Sang-Min;CHOI Soo-Il;KIM Chang-Yang;HAN Bong-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.19 no.1
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    • pp.10-19
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    • 1986
  • A study on the rapid fermentation of fish sauce has been carried out for effective utilization of sardine. The frozen sardine was thawed at room temperature, chopped, homogenized with equal amount of water and then hydrolyzed by addition of commercial proteolytic enzymes such as bromelain, papaya protease, ficin and a enzyme mixture under different conditions of hydrolysis. The effect of wheat gluten for masking fishy odor and color development during thermal treatment were also tested. The reaction mixture was heated for 30 minutes at $100^{\circ}C$ for enzyme inactivation, pasteurization and color development and then centrifuged for 20 minutes at 4,000 rpm. Finally, table salt and benzoic acid were added for bacteriostatic effect. The results were summarized as follows ; 1. The hydrolyzing temperature, time, pH and the concentration of enzymes based on the weight of whole sardine for optimal hydrolysis were as follows: autolysis, $52.5^{\circ}C$, 4 hours, pH 8.0: with $0.25\%$ bromelain, $52.5^{\circ}C$, 4 hours, pH 6.6 :with $0.25\%$ ficin, $52.5^{\circ}C$, 4 hours, pH 6.8: with $0.3\%$ papaya protease, $52.5^{\circ}C$, 4 hours, pH 6.6: with $6\%$ enzyme mixture, $52.5^{\circ}C$, 4 hours, pH 6.9, respectively. But pH control was not much beneficial in increasing yield. 2. The hydrolytic reaction of chopped sardine with proteolytic enzymes could be interpreted as a first order reaction that devided into 2 periods with different reaction rate constsnts. $Q_{10}$ values of the first period prior to 4 hours were 1.23 to 1.31, and those of post 4 hours were 1.25 to 1.55. The corresponding activation energies were $1.81{\times}10^4\;to\;2.34{\times}10^4\;kJ/kmol$ and $1.92{\times}10^4\;to\;3.77{\times}10^4\;kJ/kmol$, respectively. 3. The reasonable amount of $75\%$ vital wheat gluten for addition was $9\%$ of chopped sardine. 4. The dark brown color was mainly developed during the thermal treatment for 30 minutes at $100^{\circ}C$ and not changed during storage.

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