Characterization of a Fibrinolytic Enzyme Secreted by Bacillus velezensis BS2 Isolated from Sea Squirt Jeotgal

Zhuang, Yingping; Kim, Jeong Ah; Kim, Jeong Hwan

doi:10.4014/jmb.1810.10053

Cited by 40 publications

(32 citation statements)

References 31 publications

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“…JP-I maintained 93% residual activity after treatment for 45 min at 40°C (Figure 4b), while JP-II showed 63% residual activity after 3 min (Figure 4c). These results were similar to that of fibrinolytic enzyme from Bacillus pumilus BS15 (Yao et al, 2018), Bacillus subtilis JS2 (Yao et al, 2018), and Bacillus velezensis BS2 (Yao et al, 2019) (Table 2).…”

Section: Properties Of Purified Jp-i and Jp-iisupporting

confidence: 83%

A novel fibrinolytic enzymes from the Korean traditional fermented food—Jotgal: Purification and characterization

Kim¹,

Ri²,

Choe

2020

J Food Biochem

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The fibrinolytic activity in Korean traditional fermented food, Jotgal (pickled fish) was identified. Though the fibrinolytic activity could vary in different kinds of Jotgal, this activity seems to be produced by microorganisms during the natural fermentation stage. From Gonjaengijot (pickled opossum shrimp), two novel fibrinolytic enzymes named by JP‐I and JP‐II, have been purified by ethanol precipitation, Bio‐GEL P‐100 gel filtration, and DEAE‐cellulose ion‐exchange chromatography. Compared to the crude enzyme extract, the specific activity of the JP‐I and JP‐II increased 258, 85‐fold with the recovery of 22.1, 8.5%, respectively. The molecular weights of both enzymes were estimated as 36 kDa on sodium dodecyl sulphate‐polyacrylamide gel electrophoresis (SDS–PAGE). The optimal condition for fibrinolytic activity of JP‐I was at 50°C and pH 8.1, while that of JP‐II was at 45°C and 9.9. Both enzymes were stable at a broad range of pH (5.0 to 10.5) and have metalloprotease nature. From these results, it concludes that these enzymes could be a novel potent thrombolytic agent. Practical applications The fibrinolytic enzyme is one of the clinical agents for cardiovascular diseases which is the leading cause of morbidity and mortality worldwide with 17 million deaths every year. A variety of fibrinolytic enzymes are found and characterized from various sources such as plants, animals, and microorganisms, and new sources for fibrinolytic enzymes continue to be explored. Jotgal, widely used in Korean people's diet, is a traditional Korean seafood prepared from many different types of fishes, fish eggs, fish intestines, and shellfishes. Through an amount of research, some of fibrinolytic enzymes were found and purified from Jotgal, however, no studies have been done on fibrinolytic enzyme from opossum shrimp. In this study, the purification, enzymatic characteristics, and fibrinolytic activity of the proteases, originated from Korean traditional fermented food, Jotgal were reported. These enzymes could be novel potent thrombolytic agent.

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Section: Properties Of Purified Jp-i and Jp-iisupporting

confidence: 83%

A novel fibrinolytic enzymes from the Korean traditional fermented food—Jotgal: Purification and characterization

Kim¹,

Ri²,

Choe

2020

J Food Biochem

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“…Previous reports by our group demonstrate that OCNs effectively deliver curcumin, PNAs, and antibodies into various human and murine cells 7–10. We have also reported the use of OCNs as a delivery system for M. tuberculosis recombinant protein antigens with enhancing effect on cell-mediated immune response 18. The ability of OCNs to cross lipid bilayer membrane themselves and also to bring with them cargoes across lipid bilayer membrane was previously confirmed through both cell-sized liposomes (possess lipid bilayer membrane similar to that of real cells) and the real cells 7–9.…”

Section: Discussionmentioning

confidence: 53%

<p>Oxidized carbon nanoparticles as an effective protein antigen delivery system targeting the cell-mediated immune response</p>

Sawutdeechaikul¹,

Jiangchareon²,

Wanichwecharungruang³

et al. 2019

IJN

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Background: The demand for an effective vaccine delivery system that drives a suitable immune response is increasing. The oxidized carbon nanosphere (OCN), a negatively charged carbon nanoparticle, has the potential to fulfill this requirement because it can efficiently deliver macromolecules into cells and allows endosomal leakage. However, fundamental insights into how OCNs are taken up by antigen-presenting cells, and the intracellular behavior of delivered molecules is lacking. Furthermore, how immune responses are stimulated by OCN-mediated delivery has not been investigated. Purpose: In this study, the model protein antigen ovalbumin (OVA) was used to investigate the uptake mechanism and intracellular fate of OCN-mediated delivery of protein in macrophages. Moreover, the immune response triggered by OVA delivered by OCNs was characterized. Methods: Bone-marrow-derived macrophages (BMDMs) from mice were used to study antigen uptake and intracellular trafficking. Mice were immunized using OCN–OVA combined with known adjuvants, and the specific immune response was measured. Results: OCNs showed no cytotoxicity against BMDMs. OCN-mediated delivery of OVA into BMDMs was partially temperature independent process. Using specific inhibitors, it was revealed that intracellular delivery of OCN–OVA does not rely on phagocytosis or the clathrin- and lipid raft/caveolae-mediated pathways. Delivered OVA was found to colocalize with compartments containing MHC class I, but not with early endosomes, lysosomes, and autophagosomes. Immunization of OVA using OCNs in combination with the known adjuvant monophosphoryl lipid A specifically enhanced interferon gamma (IFNγ)- and granzyme B-producing cytotoxic T cells (CTLs). Conclusion: OCNs effectively delivered protein antigens into macrophages that localized with compartments containing MHC class I partially by the temperature independent, but not clathrin- and lipid raft/caveolae-mediated pathways. Increased CD8 + T-cell activity was induced by OCN-delivered antigens, suggesting antigen processing toward antigen presentation for CTLs. Taken together, OCNs are a potential protein antigen delivery system that stimulates the cell-mediated immune response.

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“…Interactional synergies of Saccharomyces cerevisiae and Bacillus sp., the emergence of new microorganism groups during fermentation could impact the increase and/or decrease in total polyphenols. e genus Bacillus is known for its ability to produce extracellular enzymes such as amylases [34], pectinases [35][36][37], cellulases [18,38,39], proteases [40,41], and other biomolecules as well [42]. In the context of this work, the results are promising because some identified isolates and nonidentified ones have shown degradation percentages of cellulose, amylose, and pectin up to 30% depending on the activity (cellulolytic, pectinolytic, or amylolytic) and also lysis diameters of the order of 2.2 cm ± 0.03 in the case of proteolytic activity.…”

Section: Discussionmentioning

confidence: 99%

Synergic Involvements of Microorganisms in the Biomedical Increase of Polyphenols and Flavonoids during the Fermentation of Ginger Juice

Kayath

Zamba

Mokemiabeka

et al. 2020

International Journal of Microbiology

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Steered fermentation by microorganisms gives great added value in the nutritional quality of local food. Ginger rhizome naturally contains a myriad of bioactive compounds including polyphenol and flavonoids. The aim of this work was to ferment the ginger juice, to evaluate the biochemical parameters of ginger wine, and to understand the involvement of microorganisms in the bioincrease of polyphenol compounds. Titratable acidity and pH values were determined and showed that pH is around 1.6 at the end of the fermentation when the acidity is around 6.431 g/L. Using colorimetric assay, the total polyphenolic and flavonoid compounds were evaluated throughout the fermentation. The variation of the polyphenol and flavonoid concentrations of the unsweetened sample was around 10.18 to 14.64 mg Eq AG/g and 1.394 to 2.224 mg Eq Cat/g Ms, but those from the sweet sample were around 10.82 to 18.34 mg Eq AG/g Ms and 1.311 to 2.290 mg Eq Cat/g. Using one-step PCR, multiplex techniques with specific primers, with yeast-like phenotype 27.27% (6), have been assigned among 22 isolates to Saccharomyces cerevisiae. By using PCR multiplex techniques, Bacillus licheniformis, Bacillus pumilus, Bacillus safensis, and Saccharomyces cerevisiae have been identified. Together with Saccharomyces cerevisiae, we showed that Bacillus sp. are able to secrete enzymatic landscape with some activities up to 50% including cellulase, amylase, pectinase, and protease.

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Characterization of a Fibrinolytic Enzyme Secreted by Bacillus velezensis BS2 Isolated from Sea Squirt Jeotgal

Cited by 40 publications

References 31 publications

A novel fibrinolytic enzymes from the Korean traditional fermented food—Jotgal: Purification and characterization

A novel fibrinolytic enzymes from the Korean traditional fermented food—Jotgal: Purification and characterization

<p>Oxidized carbon nanoparticles as an effective protein antigen delivery system targeting the cell-mediated immune response</p>

Synergic Involvements of Microorganisms in the Biomedical Increase of Polyphenols and Flavonoids during the Fermentation of Ginger Juice

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