• Title/Summary/Keyword: proteolytic enzyme activities

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Production of Carrot Pomace Fortified with Mucilage, Fibrinolytic Enzyme and Probiotics by Solid-state Fermentation Using the Mixed Culture of Bacillus subtilis and Leuconostoc mesenteroides

  • Jung, Hye-Won;Lee, Sam-Pin
    • Preventive Nutrition and Food Science
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    • v.14 no.4
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    • pp.335-342
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    • 2009
  • Bioactive compounds were produced from carrot pomace by solid-state fermentation using Bacillus subtilis HA and Leuconostoc mesenteroides. The carrot pomace (CP) fermented by B. subtilis HA with 3% monosodium glutamate (MSG) showed higher production of various bioactive compounds, with 1.64 Pa·sn of consistency, 2.31% of mucilage content, 16.95 unit/g of fibrinolytic enzyme activity, 35.3 unit/g of proteolytic activity and 37.5 mg% of tyrosine content. The mucilage production was greatly dependent upon the concentration of MSG added. Most MSG added in CP was converted into mucilage (2.3%) including 0.83% poly-$gamma$-glutamic acid (PGA) with 1,505 kDa of molecular weight. The CP fermented secondly by Leuc. mesenteroides showed acidic pH and lower consistency. However, the fibrinolytic and proteolytic activities were increased. The secondly fermented CP showed the viable cell counts with $2.5{\time}108$ CFU/g of B. subtilis HA and $3.7{\time}109$ CFU/g of Leuc. mesenteroides, respectively. The freeze-dried fermented CP showed 2.88 Pa·sn of consistency, 24% of mucilage content and 104.9 unit/g of fibrinolytic enzyme activity, respectively. Also, the powder of fermented CP indicated viable cell counts of $8.0{\time}107$ CFU/g of B. subtilis and $4.0{\time}108$ CFU/g of Leuc. mesenteroides. Therefore, the fermented CP that was fortified with dietary fibers, fibrinolytic enzyme and probiotics could be utilized as valuable ingredients of functional foods in food or cosmetic industries.

Extracellular proteases from bacillus licheniformis : partial purification and characterization (Bacillus licheniformis의 세포막 프로티아제 부분 정제 및 특성)

  • 홍난숙;최명언;양철학
    • Korean Journal of Microbiology
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    • v.27 no.3
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    • pp.245-249
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    • 1989
  • Extracellular proteases of Bacillus licheniformis were partially purified using ammonium sulfate fractionation and Sephadex G-75 gel filtration chromatography. The partial purification permited the weparation of two different protease activities, type I and type II. Protease type I is an enzyme with rather high protealytic activity toward dasein and was highly susceptible to organofluoride and EDTA inhibitions. It showed maximal proteolytic activity at pH 7.5 and was rapidly denatured at $71^{\circ}C$. Protease type II is a protease with relatively lower proteolytic activity than the type I. It was also inhibited by 10mM of EDTA and 1mM of PMSF by 30 min incubation. The enzyme showed maximal activity at pH 8.0 and was denatured relatively slowly at $71^{\circ}C$.

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Studies on the Microflora and Enzyme Activity in Processing of Accelerated Low Salt-Fermented Anchovy by adding koji (코오지를 이용한 속성 저식염 멸치젓의 미생물상과 효소활성)

  • 백승화;임미선;김동한
    • The Korean Journal of Food And Nutrition
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    • v.9 no.4
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    • pp.392-397
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    • 1996
  • To produce low salt fermented anchovy by an accelerated method with Asp. oryzae and Bacillus sp. koji, enzyme activity and variation of microflora during the 60 day fermentation were examined. Bacterial counts changed a little during the fermentation with the highest on day 40 for proteolytic and anaerobic bacteria and on day 20 for aerobic bacteria. Proteolytic, lipolytic, aerobic, and anaerobic bacteria counts were higher in the Bacillus sp. koji added anchovy paste than in others. The protease and lipase activities reached the highest point on day 20 and 30, respectively, and decreased gradually afterwards. The protease activity was higher in Asp. oryzae koji than in bacillus sp. koji, but the lipase activity was to the contrary.

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Relation of the Activities of Plasminogen Activator and Plasmin-like Protease with Malignant Behavior of Skin Tumor of Rats (Plasminogen Activator 및 Plasmin-like Protease활성도의 변화와 쥐 피부암의 악성)

  • Yun Kee;Park, Sang C.;Doo B. Ha;Chin H. Chung
    • The Korean Journal of Zoology
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    • v.31 no.3
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    • pp.185-190
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    • 1988
  • To investigate whether malignant behavior of skin tumor correlates with changes in the level of proteolytic activities in skin tumors, rats were treated with 7,12-dimethylbenzanthracene followed by photbor ester. Tumors induced upon the treatrnents exhibited more than 20-fold increase in the activity of plasminogen activator and about 3-fold of plasmin-like activity. as compared to those in treated controls. Furthermore, the former activity was raised to about 6-fold even in the preneoplastic dssues of the skin tissues. On the other hand, the proteolytic activity against casein and insulin decreased to several-fold in the tumor tissues while antitrvpsin activity remained similar in both tumor and controls. Thus, the increase in the activities of plasmInogen activator and plasminlike enzyme appears to occur as a charaderistic to skin cancer and may involve in invasion and metsstasis of the tumor.

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Preparation and Keeping Quality of Proteolytic Enzymes from Seafood rocessing Wastes (어류가공 부산물로부터 단백질분해 효소제의 조제 및 보관안정성)

  • KIM Jin Soo;HEU Min Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.37 no.4
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    • pp.259-268
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    • 2004
  • Keeping qualities of crude proteases (CP) and fractionated proteases (FP) sedimenting with $30\~80{\%}$ ammonium sulfate from four kinds of fish viscera as a seafood processing waste were examined. Azocaseinolytic activlties (pH 6 and 8) of CP from anchovy (Engraulis japonica), mackerel (Scomber japonicus), bastard flatfish (Pararlichthys olivaceus) and red sea bream (Chysorphys major) were stable without activity loss at $4^{\circ}C$ for 7 months. Activities of NaCP (CP containing $30{\%}$ sodium chloride) on azocasein were approximately $30{\%}$ lower than those of CP. FP activities Increased 3.4-16.1 folds compared to those of CP and NaCP Powdered crude protease (PCP) and fractionated and powdered protease (FPP) containing various sugars (lactose, sucrose, glucose and dextrin) were prepared by freeze drying. Activities of PCP and FPP containing sucrose were higher and more stable than those of PCP and FPP containing other sugars at $30^{\circ}C$ for whole keeping periods. PCP and FPP from mackerel viscera showed the highest proteolytic activity among four kind of fish vlsceras. The Optimum conditions and stabilities of FPP from mackerel viscera were pH 9 and $50^{\circ}C$, and pH 5-10 and $20-45^{\circ}C$, respectively. The results of this study suggest that FPP from seafood processing waste may be used as processing aids.

Possible Roles of Antarctic Krill Proteases for Skin Regeneration

  • Lee, Sung-Gu;Koh, Hye-Yeon;Lee, Hong-Kum;Yim, Joung-Han
    • Ocean and Polar Research
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    • v.30 no.4
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    • pp.467-472
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    • 2008
  • Antarctic krill has a strong proteolytic enzyme system, which comes from a combination of several proteases. This powerful activity can be easily detected by krill's superior post mortem autolysis. Mammalian skin consists of epidermis and dermal connective tissue, and functions as a barrier against threatening environments. A clot in a wound site of the skin should be removed for successful skin regeneration. Epithelial cells secrete proteases to dissolve the clot. In previous studies Antarctic krill proteases were purified and characterized. The proteolytic enzymes from Antarctic krill showed higher activity than mammalian enzymes. It has been suggested that these krill clean up the necrotic skin wound to induce a natural healing ability. The enzymes exhibited additional possibilities for several other biomedical applications, including dental plaque controlling agent and healing agent for corneal alkali burn. Considering that these versatile activities come from a mixture of several enzymes, discovering other proteolytic enzymes could be another feasible way to enhance the activity if they can be used together with krill enzymes. Molecular cloning of the krill proteases should be carried out to study and develop the applications. This review introduces possible roles of the unique Antarctic krill proteases, with basic information and suggestion for the development of an application to skin regeneration.

Study on the Proteolytic Activities of Pathogenic Vibrio sp. (비브리오 속의 단백질 분해능에 관한 연구)

  • 양지영;한종흔;이재우;김수광;차재호
    • Journal of Food Hygiene and Safety
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    • v.15 no.2
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    • pp.173-175
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    • 2000
  • Nine strains of pathogenic Vibrio sp. of clinical and environmental origin were examined for the degradation of gelatin, casein and hemolysin which is important to the virulence of this bacterium. Culture filtrates of all nine strains of Vibrio exhibited proteolytic activities. Especially, four strains of V. parahaemolyticus and one V. alginolyticus showed strong gelatin-degrading activity. In terms of hemolytic activity, three V. parahaemolyticus and V. mimicus showed strong $\beta$-hemolysis whereas those of strains of V. alginolyticus, V. fluvialis, V. vulnificus examined lacked this activity.

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Identification of Bacteriocin-producing Lactic Acid Bacteria from Kimchi and Partial Characterization of their Bacteriocin

  • Ha, Duk-Mo;Cha, Dong-Soo
    • Journal of Microbiology and Biotechnology
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    • v.4 no.4
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    • pp.305-315
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    • 1994
  • Nineteen strains of bacteriocin-producing lactic acid bacteria were isolated from 432 Kimchi samples, and identified by the comprehensive biochemical and morphological tests verifying their cellular fatty acid composition. Using partially purified bacteriocins from these isolates, their inhibitory activities against other lactic acid bacteria and some pathogens, and sensitivity to enzyme and heat treatments were tested. The isolates were identified as Lactobacillus plantarum (2 strains), L curvatus (2 starins), L brevis (2 strains), Enterococcus faecium (6 strains), Leuconostoc mesenteroides subsp. mesenteroides (1 strain) and Lactobacillus sp. (6 strains). The bacteriocins produced by E. faecium strains provided the broadest spectrum of inhibition, affecting against other Gram-positive bacteria including lactic acid bacteria and health-threatening bacteria such as Clostridium perfringens and Listeria monocytogenes. The bacteriocins of Lactobacillus sp., L plantarum and L brevis strains were capable of inhibiting many strains of the lactic acid bacteria, whereas those of L curvatus and L mesenteroides subsp. mesenteroides strains were only inhibitory to a few strains. Generally, the inhibitory activities of both E. faecium and Lactobacillus sp. strains were greater than those of other producer strains. The bacteriocins from the isolates were sensitive to several proteolytic enzymes, and those of L curvatus and L mesenteroides subsp. mesenteroides were also sensitive to lipase and $\alpha$-amylase as well as to proteolytic enzymes. The bacteriocins from the strains of Lactobacillus sp. and a strain of L. brevis were resistant to autoclaving.

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Studies on Protein Contents and Enzyme Activities of Human Seminal Plasma (사람정장(精漿)의 단백질농도(蛋白質濃度)와 효소활성도(酵素活性度)에 관한 연구(硏究))

  • Park, Kyu-Hong;Lee, Hee-Yong
    • Clinical and Experimental Reproductive Medicine
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    • v.10 no.1
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    • pp.7-24
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    • 1983
  • On the basis of the semen analysis in 66 subjects, they were divided into six different groups: Group I consisted of 16 normal subjects with sperm counts of over 40 ${\times}10^6$/ml and motility of over 40 percent, Group II, 7 subjects with normal sperm counts, but motility of under 40 percent, Group III, 15 oligospermic patients with under 40 ${\times}10^6$/ml, Group IV 14 azoospermic patients, Group V, 10 patients with vasectomy and Group VI, 4 abnormal patients with 2 cases of hypoplastic testis, 1 case of Klinefelter's syndrome and 1 case of testis tumor. After seperation of semen into sperm and seminal plasma by centrifugation, the protein contents and the activities of hyaluronidase, ${\beta}$-N acetylglucosaminidase, ${\beta}$-glucuronidase, arylsulfatase, acrosin and azocoll proteinase in seminal plasma were measured. Vasectomy group has 30 percent less of total protein than normal group. For the comparison of enzyme activities of seminal plasma, it could be assumed that the enzymes in seminal plasma were not contaminated with the enzymes of spermatozoa by testing the enzymes of the seminal plasma from the vasectomy and azoospermic groups. It had been reported that hyaluronidase was only released from spermatozoa, however, the result obtained in this investigation showed that azoospermic and vasectomy group had high specific activities of hyaluronidase. The results indicated that hyaluronidase was not only from the testis but also from the male accessory sexual glands. Oligospermic group (Group III) showed the lowest total activity of hyaluronidase among them. The specific activities of ${\beta}$ -N-acetylglucosaminidase was high in oligospermic group (Group III) and low in vasectomy group (Group V). These results were contradictory with the pattern of hyaluronidase activities. This indicated that the spermatozoa which were stayed in epididymis would increase the activity of this enzyme. The specific activity of ${\beta}$ - glucuronidase was low in oligospermic and vasectomy groups. Group VI including testis tumor had remarkably high arylsulfatase activity. Arylsulfatase, a typical lysosomal enzyme, has been known to be released unusually large amounts from certain tumor cells. Arylsulfatase was also released with high activities from azoospermic and vascetomy group. This result indicated that this enzyme was also released from the sources other than testis. Acrosin, a proteolytic enzyme locating in the sperm acrosome, was not found throughout all the samples of seminal plasma. The activities of azocoll proteinase, a non-specific neutral proteinase was nearly identical in all the groups. This enzyme must have been released from the sources other than testis.

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Screening and Characterization of Potential Bacillus Starter Cultures for Fermenting Low-Salt Soybean Paste (Doenjang)

  • Jeon, Hye Hee;Jung, Ji Young;Chun, Byung-Hee;Kim, Myoung-Dong;Baek, Seong Yeol;Moon, Ji Young;Yeo, Soo-Hwan;Jeon, Che Ok
    • Journal of Microbiology and Biotechnology
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    • v.26 no.4
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    • pp.666-674
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    • 2016
  • The bacterial strains were screened as potential starters for fermenting low-salt doenjang (a Korean traditional fermented soybean paste) using Korean doenjang based on proteolytic and antipathogenic activities under 6.5-7.5% NaCl conditions. Phylogenetic analysis based on 16S rRNA gene sequences showed that they all belonged to the genus Bacillus. Proteolytic and antipathogenic activities against Escherichia coli, Bacillus cereus, Staphylococcus aureus, Listeria monocytogenes, and Aspergillus flavus, as well as fibrinolytic, amylase, and cellulase activities of the 10 strains were quantitatively evaluated. Of these, strains D2-2, JJ-D34, and D12-5 were selected, based on their activities. The functional, phenotypic, and safety-related characteristics of these three strains were additionally investigated and strains D2-2 and D12-5, which lacked antibiotic resistance, were finally selected. Strains D2-2 and D12-5 produced poly-γ-glutamic acid and showed various enzyme activities, including α-glucosidase and β-glucosidase. Growth properties of strains D2-2 and D12-5 included wide temperature and pH ranges, growth in up to 16% NaCl, and weak anaerobic growth, suggesting that they facilitate low-salt doenjang fermentation. Strains D2-2 and D12-5 were not hemolytic, carried no toxin genes, and did not produce biogenic amines. These results suggest that strains D2-2 and D12-5 can serve as appropriate starter cultures for fermenting low-salt doenjang with high quality and safety.