• Microbiology and Biotechnology Letters
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• v.51 no.2
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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.

• Journal of Microbiology and Biotechnology
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• v.19 no.9
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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.

• Korean Journal of Agricultural Science
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• v.49 no.4
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• pp.995-1001
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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.

• Applied Chemistry for Engineering
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• v.19 no.1
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• pp.122-128
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• 2008

We evaluated the relationship between the multiplication of heterotrophic microorganisms and physicochemical factors in the final discharged sewage water from J municipal waste water treatment plants. Dissolved organic carbon (DOC) was the most crucial factor influencing multiplication of heterotrophic plate counts (HPC) among the water quality variables selected. Degrading bacteria, such as proteolytic bacteria, lipholytic bacteria, starch degrading bacteria, cellulolytic bacteria, and pectinolytic bacteria, were monitored to understand the condition of nutrients in finished sewage effluent. The percentages of lipid and protein combined occupied 81% in finished sewage water. The multiplication of HPC showed the highest value in August. The formation of trihalomethane (THM) was low in the finished discharge water during chlorine disinfection, which was $71{\mu}/L$ (which was less than $100{\mu}/L$- the standard of drinking water quality) with 10 mg/L of chlorine during 15 min.

• Microbiology and Biotechnology Letters
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• v.37 no.1
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• pp.24-32
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• 2009

The KNUC25 strain isolated from over-fermented whole Chinese cabbage kimchi was examined for its physiological characteristics using API 50 CHL system assay and identified as Lactobacillus paraplantarum by analysis of whole-cell protein SDS-PAGE pattern assay and similarity of 16S rDNA sequence. L. paraplantarum KNUC25 had a broad antimicrobial activity spectrum from Gram positive to Gram negative bacteria. Scanning electron micrograph analysis showed that KNUC25 might attack to cell surface of indicator cells and destruction can lead to inhibition of the cell growth. The antimicrobial substance of the KNUC25 strain was stable to various degrading enzymes and at high temperature and not a plasmid-born matter. Resistance to proteolytic enzymes showed that an antimicrobial activity of KNUC25 might not be caused by proteinous substance. Maximum production of antimicrobial substance was the exponential growth phase at $30^{\circ}C$.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.12
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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.