• 제목/요약/키워드: proteolytic indicator

검색결과 6건 처리시간 0.024초

Action of Extracellular Protease of Aspergillus terreus on Human Plasma Hemostasis Proteins

  • Alexander A. Osmolovskiy;Elena S. Zvonareva;Nina A. Baranova;Valeriana G. Kreyer
    • 한국미생물·생명공학회지
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    • 제51권2호
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    • pp.167-173
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    • 2023
  • Proteolytic enzymes secreted by Aspergillus, as pathogenicity factors, affect blood coagulation and fibrinolysis, and therefore the target proteins of their action in the bloodstream are of significant interest. In the present study, the action of the isolated protease of A. terreus 2 on different human plasma proteins was shown. The protease of A. terreus 2 exhibited the highest proteolytic activity against hemoglobin, which was 2.5 times higher than the albuminolytic activity shown in both of the protein substrates used. In addition, the protease has significant ability to hydrolyze both fibrin and fibrinogen. However, the inability of the A. terreus 2 protease to coagulate rabbit blood plasma and coagulate human and bovine fibrinogen indicates the severity of the enzyme's action on human blood coagulation factors. It should be considered as a potential indicator of this isolated protease's participation in fungal pathogenesis. The protease shows no hemolytic activity. Furthermore, its activity is insignificantly inhibited by thrombin inhibitors, and is not inhibited by plasmin inhibitors.

Monitoring of Cleavage Preference for Caspase-3 Using Recombinant Protein Substrates

  • Park, Kyoung-Sook;Yi, So-Yeon;Kim, Un-Lyoung;Lee, Chang-Soo;Chung, Jin-Woong;Chung, Sang-J.;Kim, Moon-Il
    • Journal of Microbiology and Biotechnology
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    • 제19권9호
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    • pp.911-917
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    • 2009
  • The apoptotic caspases have been classified in accordance with their substrate specificities, as the optimal tetrapeptide recognition motifs for a variety of caspases have been determined via positional scanning substrate combinatorial library technology. Here, we focused on two proteolytic recognition motifs, DEVD and IETD, owing to their extensive use in cell death assay. Although DEVE and IETD have been generally considered to be selective for caspase-3 and -8, respectively, the proteolytic cleavage of these substrates does not display absolute specificity for a particular caspase. Thus, we attempted to monitor the cleavage preference for caspase-3, particularly using the recombinant protein substrates. For this aim, the chimeric GST:DEVD:EGFP and GST:IETD:EGFP proteins were genetically constructed by linking GST and EGFP with the linkers harboring DEVD and IETD. To our best knowledge, this work constitutes the first application for the monitoring of cleavage preference employing the recombinant protein substrates that simultaneously allow for mass and fluorescence analyses. Consequently, GST:IETD:EGFP was cleaved partially in response to caspase-3, whereas GST:DEVD:EGFP was completely proteolyzed, indicating that GST:DEVD:EGFP is a better substrate than GST:IETD:EGFP for caspase-3. Collectively, using these chimeric protein substrates, we have successfully evaluated the feasibility of the recombinant protein substrate for applicability to the monitoring of cleavage preference for caspase-3.

Comparison of changes in functional characteristics of fermented soybean with different microbial strains

  • Hyewon Lim;Bosung Kim;Heewon Jung;Sungkwon Park
    • 농업과학연구
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    • 제49권4호
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    • pp.1047-1053
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    • 2022
  • The purpose of this study was to compare the effect of solid-state fermentation on soybean using three microbial strains under four different fermentation times. Soybean was fermented for 12, 24, 36 or 48 hours with highly proteolytic microbes, either Bacillus amyloliquefaciens (BA), B. subtilis (BS), or B. subtilis var. natto (BN), and levels of total protein concentration, protein distribution, and antioxidant activity were analyzed. Total protein was highest in the BS 12 h group (9.21 ㎍·µL-1) and lowest in BN 48 h (6.80 ㎍·µL-1), respectively (p < 0.001). Furthermore, three microbes decomposed large molecular weight proteins as well as major allergens of soybean such as β-conglycinin, Gly m Bd 30K, and glycinin. Each treatment group showed the highest degradation rate at 48 h fermentation and among the three microbes, BS showed a relatively higher degradation rate. The radical scavenging ability, known as an indicator of antioxidant activity, showed a significant increase in all treatment groups except BA 24 h. The results from this study suggest that protein concentration, and degradation and antioxidant activity were affected by different types of microbial trains and fermentation period and that B. subtilis fermentation might be the most effective way to increase nutritional and functional properties of soybean.

J 하수 처리장 방류수 중 세균의 성장 거동 및 염소 소독 효율 고찰 (Study on Microorganism Multiplication Behavior and Efficiency of Chlorine Disinfection in the Sewage Effluent from J Municipal Waste Water Treatment Plant)

  • 이운기;이윤진;정규연
    • 공업화학
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    • 제19권1호
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    • pp.122-128
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    • 2008
  • 본 연구에서는 J 하수 처리장의 최종 방류수를 대상으로 종속영양세균의 증식과 수중 물리, 화학적 영향인자와의 관계를 고찰하였다. 하수 중의 용존 유기물질 농도는 종속영양세균의 성장과 가장 밀접한 관계를 보였다. 미생물의 영양원인 단백질, 지방, 전분, 섬유소, 팩틴의 분해 세균 분포를 파악한 결과 하수 중 단백질 및 지방세균의 비율은 전체의 81%로 가장 높았다. 종속영양세균은 연중 8월에 가장 높은 수치를 나타냈다. 하수 방류수에 대해 10 mg/L의 유리염소를 주입하여 15 min간 접촉시 THMs의 농도는 $71{\mu}/L$ (< $100{\mu}/L$- 먹는 물 수질기준)의 값을 나타내어 높지 않았다.

김치로부터 분리된 Lactobacillus paraplantarum KNUC25가 만드는 항균 물질의 특성 (Characterization of Antimicrobial Substance Produced by Lactobacillus paraplantarum KNUC25 Isolated from Kimchi)

  • 김마리;이수진;설경조;박유미;김사열
    • 한국미생물·생명공학회지
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    • 제37권1호
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    • pp.24-32
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    • 2009
  • 숙성 정도가 오래된 김장 배추 김치에서 분리되어 16S rDNA 염기 서열 분석을 통해 부분 동정된 KNUC25 분리 균주를 단백질 전기 영동 패턴과 생리적 특징 그리고 염기 서열의 유사성을 비교하여 Lactobacillus paraplantarum로 동정하였다. L. paraplantarum KNUC25의 농축 상등액은 그람 양성균과 음성균에 넓은 범위의 항균 활성을 나타냈다. 전자 현미경을 통해 KNUC25가 만들어내는 항균 물질은 세균의 표면에 작용하여 생육을 억제하는 것으로 확인되었다. 항균 활성 물질을 생산하는 최적 온도는 섭씨 30도이고, 높은 온도에서도 항균 활성을 보였으며 단백질 분해 효소에 안정하며 비단백질성 항균 물질일 가능성을 보여주었다.

Clinicopathological and Prognostic Significance of MUC-2, MUC-4 and MUC-5AC Expression in Japanese Gastric Carcinomas

  • Xiao, Li-Jun;Zhao, Shuang;Zhao, En-Hong;Zheng, Xin;Gou, Wen-Feng;Xing, Ya-Nan;Takano, Yasuo;Zheng, Hua-Chuan
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권12호
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    • pp.6447-6453
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    • 2012
  • Background: The mucin components of the gastric gel layer function as a protective and lubricating factor against luminal acid and proteolytic enzymes. Alteration of mucin expression in gastric preneoplastic and neoplastic lesions has suggested potential roles in neoplastic processes. This study aimed to assess the clinicopathological and prognostic significance of MUC-2, MUC-4 and MUC-5AC in Japanese gastric cancer. Methods: Expression of MUC-2, -4 and -5AC was evaluated on tissue microarrays of gastric carcinomas and adjacent non-cancerous mucosa specimens by immunohistochemistry and compared with clinicopathological parameters and survival time of the patients. Results: The three mucins were found to be expressed to a lesser extent in gastric carcinomas in comparison with non-cancerous mucosa (p<0.05). MUC-2 expression was negatively correlated with tumor size, depth of invasion, and TNM staging of gastric cancer (p<0.05), while that of MUC-5AC was negatively associated with the depth of invasion, venous invasion, lymph node metastasis and TNM staging (p<0.05), but positively with MUC-4 and MUC-2 expression (p<0.05). There was higher MUC-2 expression in intestinal- than diffuse-type carcinomas (p<0.05). Kaplan-Meier analysis indicated no relationship between expression of the three mucins and the cumulative survival rate of patients, even stratified according to the depth of invasion (p>0.05). Conclusion: Down-regulated expression of MUC-2, -4 and -5AC may be involved in pathogenesis, invasion, metastasis or differentiation of gastric carcinoma. Their altered expression might therefore be employed as an indicator of pathobiological behavior.