• Title/Summary/Keyword: proteolytic pattern

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Characteristics of the Bacteriocin from Lactobacillus sp. Oh-B3. (Lactobacillus sp. Oh-B3로부터 생산되는 박테리오신의 특성)

  • 김동섭
    • Microbiology and Biotechnology Letters
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    • v.30 no.2
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    • pp.184-188
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    • 2002
  • A bacteriocin producing microorganism, which inhibits the growth of Lactobacillus sake, was screened and isolated from Kimchi. This microorganism was identified and named as Lactobacillus sp. Oh-B3, The maximum amount of bacteriocin was produced when the isolated microorganism was cultured in MRS media(pH 8.0) for 24 hours at 25℃. The bacteriocin from the isolated microorganism was purified through ammonium sulfate precipitation, dialysis and ultrafiltration. The bacteriocin was stable on the wide pH range of 2.0-9.0, and showed antimicrobial activity on some of gram positive bacteria, not on gram negative. The antimicrobial activity of bacteriocin was mostly removed by treatment of proteolytic enzymes. But, the bacteriocin was very stable on the heat treatment, and more than 50% of activity was remained at autoclaving. The action mode of the bacteriocin showed bacteriocidal pattern, being same as that of general bacteriocins.

Characteristics and Action Pattern of Alkaline Protease produced from Aspergillus fumigatus (Aspergillus fumigatus이 생산하는 Alkaline protease의 특성과 작용양상)

  • Cha, Woen-Suep;Choi, Cheong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.18 no.3
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    • pp.348-355
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    • 1989
  • This experiment was conducted to investigate the characteristics of alkaline protease from Aspergillus fumigatus which was isolated from soil as a superior strain for the production of the alkaline protease. The optimum temperature for enzyme activity was $50^{\circ}C$ and optimum pH was 9.0. The enzyme was stable at pH 8.0 to 10.0 and thermal inactivation was shown $30^{\circ}C$. The activity of the enzyme was increased by the addition of $Mn^{++},\;Cu^{++},\;Ba^{++},\;Mg^{++},\;$wheras it was inhibitied by $K^+,\;Fe^{+++},\;Ag^+,\;Pb^{++},\;Na^+,\;Ca^{++},\;Hg^+,\;Zn^{++}$. EDTA. 2, 4-DNP, ${\varepsilon}-amino$ caproic acid did not show inhibitory effect on the proteolytic activity of alkaline protease but P-chloromercuribenzoic acid inhibited the enzyme activity, indicating that reactive sulfhydryl group is required for the enzymatic activity. The reaction of this enzyme followed typical Michael-Menten Kinetics with the Km value of $8.33{\times}10^{-4}mole/{\ell}$ with the Vmax of $47.62{\mu}g/min$. This enzyme had stronger proteolytic activity than trypsin on substrate such as casin and hemoglibin.

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Distribution and Activity of Hheterotrophic Bacteria in the Mudflat of Nakdong River Estuary (난동감 하구 간석지에 존재하는 세균의 분포 및 생리적 활성도)

  • Kim, Sang-Jong;Hong, Soon-Woo;Rhie, Youn;Choi, Sung-Chan
    • Korean Journal of Microbiology
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    • v.23 no.3
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    • pp.215-222
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    • 1985
  • Distribution pattern and activity of heterotrophec bacteria were measured in the mudflat of Nakdong river estuary. In March and June, 1985, community sizes of amylolytic, lipolytic and proteolytic bacteria as well as total viable counts were measured. Vertical distribution of bacterial community size increased a few orders of magnitude from Narch to June. Heterotrophic activity was estimated in turnover time with $U-[^{14}C]-glucose$. Turnover time reduced considerably in June compared to that of March. To sxamine correlations for measured bacterial groups, turnover time and environmental factors, correlation coefficient matrix was obtained. These measured characteristics did not consistently correlate well with one another.

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Occurrence of Thioredoxin Reductase in Deinococcus Species, the UV resistant Bacteria

  • Seo Hee-Jeong;Lee Young-Nam
    • Journal of Microbiology
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    • v.44 no.4
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    • pp.461-465
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    • 2006
  • The occurrence of thioredoxin reductase (NAD(P)H: oxidized-thioredoxin reductase, EC 1.6.4.5, TrxR) in five mesophilic species of Deinococcus was investigated by PAGE. Each species possessed a unique TrxR pattern, for example, a single TrxR characterized D. radiopugnans while multiple forms of TrxR occurred in other Deinococcal spp. Most of TrxRs occurring in Deinococcus showed dual cofactor specificity, active with either NADH or NADPH, although the NADPH specific-TrxR was observed in D. radiophilus and D. proteolytic us.

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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A Study on the Proteolysis of Mussel Protein by a Commercial Enzyme Preparation (단백질 분해효소에 의한 홍합 단백질의 분해에 관한 연구)

  • Choi, In-Jae;Nam, Hee-Sop;Shin, Zae-Ik;Lee, Byong-Hoon
    • Korean Journal of Food Science and Technology
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    • v.24 no.6
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    • pp.519-523
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    • 1992
  • The patterns on the proteolysis of mussel protein using a commercial enzyme preparation were investigated. The best one among six commercial enzyme preparations for the manufacture of mussel extract was Corolase PP, based on the degree of hydrolysis (DH). When the raw mussel paste, without water addition, was adjusted to pH 6.5, added 0.1% (w/w dry basis) of Corolase PP. and reacted at $50^{\circ}C$ for four hours, it reached the maximum value of DH (79%). The precooking of raw mussel decreased the efficiency of extraction and hydrolysis of the protein, due to the inactivation of the autolytic enzymes contained in the mussel. During the course of proteolysis, major free amino acids such as glycine, alanine, glutamic acid and lysine, representing a characteristic brothy taste of mussel were replaced with free hydrophobic amino acids including valine, methionine, isoleucine, and leucine. The electrophoretic pattern and HPLC-GPC pattern of mussel protein hydrolysates during the hydrolysis were observed and also discussed.

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Molecular Cloning and Expression Pattern of Abalone (Haliotis discus hannai) Myostatin cDNA (참전복(Haliotis discus hannai) Myostatin 유전자의 cDNA 동정 및 발현 분석)

  • Lee, Sang-Beum;Kim, Jeong-Hwan;Jo, Mi-Jin;Oh, Mi-Young;Park, Heum-Gi;Jin, Hyung-Joo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.42 no.2
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    • pp.139-145
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    • 2009
  • We cloned and sequenced the open reading frame (ORF) cDNA encoding myostatin from the muscle of abalone (Haliotis discus hannai). The ORF cDNA of the abalone myostatin is 1134 bp and encoded 377 amino acid residues that were 60-96% homologous with the amino acids of other organism myostatins. In addition, the ORF contained a conserved proteolytic cleavage site (RXRR) and nine conserved cysteine residues in the C-terminus. Semi-quantitative RT-PCR revealed the presence of myostatin mRNA in various tissues. The strongest expression was observed in the mantle of female abalone, and the gills and heart of male abalone.

Apoptosis Induction by Menadione in Human Promyelocytic Leukemia HL-60 Cells

  • Sa, Duck-Jin;Lee, Eun-Jee;Yoo, Byung-Sun
    • Toxicological Research
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    • v.25 no.3
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    • pp.113-118
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    • 2009
  • Cell death induced by menadione (vitamin K-3,2-methyl-1,4-naphthoquinone) has been investigated in human promyelocytic leukemia HL-60 cells. Menadione was found to induce both apoptosis and necrosis in HL-60 cells. Low concentration ($1{\sim}$50 ${\mu}$M) of menadione induced apoptotic cell death, which was demonstrated by typical DNA ladder patterns on agarose gel electrophoresis and flow cytometry analysis. In contrast, a high concentration of menadione (100 ${\mu}$M) induced necrotic cell death, which was demonstrated by DNA smear pattern in agarose gel electrophoresis. Necrotic cell death was accompanied with a great reduction of cell viability. Menadione activated caspase-3, as evidenced by both increased protease activity and proteolytic cleavage of 116 kDa poly(ADP-ribose) polymerase (PARP) into 85 kDa cleavage product. Caspase-3 activity was maximum at 50 ${\mu}$M of menadione, and very low at 100 ${\mu}$M of menadione. Taken together, our results showed that menadione induced mixed types of cell death, apoptosis at low concentrations and necrosis at high concentrations in HL-60 cells.

Ablation of Arg-tRNA-protein transferases results in defective neural tube development

  • Kim, Eunkyoung;Kim, Seonmu;Lee, Jung Hoon;Kwon, Yong Tae;Lee, Min Jae
    • BMB Reports
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    • v.49 no.8
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    • pp.443-448
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    • 2016
  • The arginylation branch of the N-end rule pathway is a ubiquitin-mediated proteolytic system in which post-translational conjugation of Arg by ATE1-encoded Arg-tRNA-protein transferase to N-terminal Asp, Glu, or oxidized Cys residues generates essential degradation signals. Here, we characterized the ATE1−/− mice and identified the essential role of N-terminal arginylation in neural tube development. ATE1-null mice showed severe intracerebral hemorrhages and cystic space near the neural tubes. Expression of ATE1 was prominent in the developing brain and spinal cord, and this pattern overlapped with the migration path of neural stem cells. The ATE1−/− brain showed defective G-protein signaling. Finally, we observed reduced mitosis in ATE1−/− neuroepithelium and a significantly higher nitric oxide concentration in the ATE1−/− brain. Our results strongly suggest that the crucial role of ATE1 in neural tube development is directly related to proper turn-over of the RGS4 protein, which participate in the oxygen-sensing mechanism in the cells.

Antagonism against Helicobacter Pylori and Proteolysis of Lactobacillus Helveticus CU631 and Strain Identification

  • Yoon, Y.H.;Won, B.R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.7
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    • pp.1057-1065
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    • 2002
  • The antagonistic activities of 30 strains of lactobacilli against Helicobacter pylori were determined and Lactobacillus helveticus CU631 has been selected as the strain which possesses the strongest inhibitory effect in the disc diffusion assay showing inhibition zone diameter of $10{\pm}1.5mm$, whereas those of L. plantarum and L. fermentum have been shown to be $4.0{\pm}0.6mm$. H. pylori G88016 revealed the highest vacuolating toxin producing activity among the 8 strains, the inhibitory activity of L. helveticus CU631 in vacuolating toxin producing activity of H. pylori manifested in the co-culture of two strains and in the 5:5 mixture of supernatant of the two strains. Both L. helveticus CU631 and cell free culture supernatant had a strong inhibitory activities in urease and cytotoxin producing activities of H. pylori NCTC11637 and CJH12. An accelerated proteolytic activity of water soluble peptides by L. helveticus CU631 during the refrigeration storage has been manifested in the cream cheese. DNA seqences of 16S-23S ribosomal RNA spacer region showed typical pattern among the various strains of L. helveticus, which could be used in the identification of L. helveticus CU 631.