• Advances in Traditional Medicine
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• v.6 no.4
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• pp.274-285
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• 2006

Allium sativum (Family Amaryllidaceae or Liliaceae) is used worldwide for various clinical uses like hypertension, cholesterol lowering effect, antiplatelets and fibrinolytic activity etc. Due to these common house hold uses of Allium sativum, as a herbal supplements, and failure of patients to inform their physician of the over-the-counter supplements they consume leads to drugnutrient interactions with components in herbal supplements. Today these types of interactions between a herbal supplement and clinically prescribed drugs are an increasing concern. In vitro studies indicated that garlic constituents modulated various CYP (cytochrome P450) enzymes. CYP 3A4 is abundantly present in human liver and small intestine and contributes to the metabolism of more than 50% of commonly used drugs including nifedipine, cyclosporine, erythromycin, midazolam, alprazolam, and triazolam. Extracts from fresh and aged garlic inhibited CYP 3A4 in human liver microsomes. The in vivo effects of garlic constituents are found to be species depended and the dosing regimen of garlic constituents appeared to influence the modulation of various CYP isoforms. Studies have indicated that the inhibition of various CYPs by organosulfur compounds from garlic was related to their structure also. Studies using in vitro, in vivo, animal and human models have indicated that various garlic constituents can be the substrates, inhibitors and or inducers of various CYP enzymes. The modulation of CYP enzyme activity and expression are dependent on the type and chemical structure of garlic constituents, dose regime, animal species and tissue, and source of garlic thus this review throws light on the possible herb drug interaction with the use of garlic.

• Journal of Life Science
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• v.14 no.3
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• pp.509-513
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• 2004

Inhibitory activities of 58 methanol extracts, which were prepared from different parts of 55 kinds of edible and medicinal plants, against human thrombin were evaluated. Analysis of the anti-coagulation activity determined by thrombin time enabled us to select 8 herb extracts as possible sources of anti-thrombosis agent. Further analysis of anti-thrombosis activity determined by activated partial thromboplastin time and fibrinolytic activity, and heat stability and inhibition against other proteolytic digestive enzymes provided new information that Pimpinella brachycarpa, Lysimachia clethroides, and Salix gracilistyla could be used as a potential anti-thrombosis agent. Our results suggest that edible and medicinal plants could be the potential source of thrombin inhibitor.

• BMB Reports
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• v.37 no.5
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• pp.574-581
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• 2004

The full-length cDNA of the lumbrokinase fraction 6 (F6) protease gene of Lumbricus rubellus was amplified using an mRNA template, sequenced and expressed in E. coli cells. The F6 protease gene consisted of pro- and mature sequences by gene sequence analysis, and the protease was translated and modified into active mature polypeptide by N-terminal amino acid sequence analysis of the F6 protease. The pro-region of F6 protease consisted of the 44 residues from methionine-1 to lysine-44, and the mature polypeptide sequence (239 amino acid residues and one stop codon; 720 bp) started from isoleucine-45 and continued to the terminal residue. F6 protease gene clones having pro-mature sequence and mature sequence produced inclusion bodies in E. coli cells. When inclusion bodies were orally administrated rats, generated thrombus weight in the rat' venous was reduced by approximately 60% versus controls. When the inclusion bodies were solubilized in pepsin and/or trypsin solutions, the solubilized enzymes showed hemolytic activity in vitro. It was concluded the F6 protease has hemolytic activity, and that it is composed of pro- and mature regions.

• BMB Reports
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• v.35 no.2
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• pp.236-238
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• 2002

Based on the zymography analysis, Bacillus sp. DJ-4 (screened from Doen-Jang, a Korean traditional fermented food) secretes seven extracellular fibrinolytic enzymes (EFEs; 68, 64, 55, 45, 33, 27, and 13 kDa) in culture broth. These seven EFEs were analyzed by newly applied SDS-fibrin zymography combined with gradient polyacrylamide (SDS-FZGP). This improved gel system was used with a 5-20% acrylamide gradient in a fibrin zymogram gel for the separation of proteins with molecular masses from below 10kDa to over 100kDa on one gel plate. Using this system, high molecular weight bands (HMWBs) were clearly and sharply resolved. We also examined the relationship between an acrylamide concentration and the enzymatic activity of EFE using densitometric analysis.

• Korean Journal of Pharmacognosy
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• v.35 no.1 s.136
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• pp.52-61
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• 2004

Inhibitory activities of 264 methanol extracts, which were prepared from different parts of 210 kinds of wild and medicinal plants, against human thrombin were evaluated. Based on the anti-coagulation activity determined by thrombin time and activated partial thromboplastin time, the 14 extracts were screened. The fibrinolytic activity, heat stability and inhibition of other proteolytic digestive enzymes, such as pepsin, papain, trypsin and chymotrypsin, of the 14 extracts were further determined, and Ginko biloba (herba), Ephedra sinica (radix), Reynoutria elliptica (herba), Amomum tsao-ko Crevost (fructus), and Magnolia officinalis Rehd. et Wils (bark) were finally selected as possible plant sources for anti-thrombosis agent. These results suggested that medicinal and wild plants could be the potential source of thrombin inhibitor.

• Microbiology and Biotechnology Letters
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• v.38 no.4
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• pp.379-384
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• 2010

We isolated microorganisms presenting high enzymatic activities for amylase, cellulase, protease, lipase or fibrinolysis from Korean traditional soybean sauce and paste. Then, the physiological properties and 16S rRNA sequences of isolated microorganisms were analyzed. All of the isolated 13 strains possessing high extra cellular enzyme activities have higher amylase and cellulase activities than Bacillus subtilis KACC 10114. All the selected strains have protease activities except for D2-14. Except D8-8 and K4-1, other strains have lipase activity. D2-7, D8-8 and K4-1 strains have higher fibrinolytic activities than others, while D8-2 strain has no activity. Most of the selected strains showed antibacterial activity even in gram positive and gram negative bacteria and yeast. Gene sequence analysis of 16S rRNA from isolated strains revealed that all the selected strains were member of Bacillus species.

• BMB Reports
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• v.37 no.3
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• pp.298-303
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• 2004

In general, a SYPRO Ruby dye is well known as a sensitive fluorescence-based method for detecting proteins by one-or two-dimensional SDS-PAGE (1-DE or 2-DE). Based on the SYPRO Ruby dye system, the combined two-dimensional fibrin zymography (2-D FZ) with SYPRO Ruby staining was newly developed to identify the Bacillus sp. proteases. Namely, complex protein mixtures from Bacillus sp. DJ-4, which were screened from Doen-Jang (Korean traditional fermented food), showed activity on the zymogram gel. The gel spots on the SYPRO Ruby gel, which corresponded to the active spots showing on the 2-D FZ gel, were analyzed by a matrix-assisted laser desorption ionization time of flight (MALDI-TOF) mass spectrometric analysis. Five intracellular fibrinolytic enzymes of Bacillus sp. DJ-4 were detected through 2-D FZ. The gel spots on the SYPRO Ruby dye stained 2-D gel corresponding to 2-D FZ were then analyzed by MALID TOF MS. Three of the five gel spots proved to be quite similar to the ATP-dependent protease, extracellular neutral metalloprotease, and protease of Bacillus subtilis. Also, the extracellular proteases of Bacillus sp. DJ-4 employing this combined system were identified on three gels (e.g., casein, fibrin, and gelatin) and the proteolytic maps were established. This combined system of 2-D zymography and SYPRO Ruby dye should be useful for searching the specific protease from complex protein mixtures of many other sources (e.g., yeast and cancer cell lines).

• Journal of the Korean Society of Food Science and Nutrition
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• v.36 no.8
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• pp.1070-1076
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• 2007

Soymilk cake (SMC) fortified with defatted soy flour was fermented by a novel Bacillus subtilis GT-D in the solid-state fermentation with chlorella, mulberry leaves or green tea as food ingredient. The fermentation of SMC resulted in higher content of tyrosine and higher activities of protease, amylase and fibrinolytic enzymes. Fermented SMC including mucilage indicated six-fold higher content of dietary fiber compared to that of non-fermented SMC. The SMC fermented without additional ingredient allowed the highest spread ratio and width of cookies. Cookies prepared without SMC showed the lowest value of spread ratio and width compared to those of fermented SMC. In addition, these cookies showed the highest hardness value of 8,975 g, and thickness value of 5.97 and slightly higher bright color. Cookies fortified with SMC fermented with chlorella or mulberry leaves showed higher sensory evaluation value. Fortification of SMC fermented without additional ingredient resulted in the highest preference in the sensory evaluation of cookies.