• Microbiology and Biotechnology Letters
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• v.40 no.1
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• pp.57-65
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• 2012

Antifungal compounds from Bacillus polyfermenticus CJ6 were purified using SPE, preparative HPLC, and reverse phase-HPLC. Antifungal compounds from B. polyfermenticus CJ6 were separated into three fractions (8, B, C) using preparative HPLC. LC/MS analysis of antifungal peaks suggested that B. polyfermenticus CJ6 produces lipopeptides; two kinds of iturin A ($C_{14}$, $C_{15}$), three kinds of surfactins ($C_{13}$, $C_{14}$, $C_{15}$), four kinds of fengycin A ($C_{14}$, $C_{15}$, $C_{16}$, $C_{17}$) and two kinds fengycin B ($C_{16}$, $C_{17}$). The antifungal activity of fraction 8, which was presumed as inturin A, was found to be stable after the pH, heat or proteolytic enzyme treatment, but it was unstable at 50-$70^{\circ}C$ for 24 hr. The antifungal activity of fraction B, which presumed as surfactins and fengycin A, was found to be stable after the heat treatment, but it was unstable in the pH 3.0 and after the protease (type I) or ${\alpha}$-chymotrypsin treatment. The antifungal activity of fraction C, which was presumed as fengycin A and B, was found to be stable in the pH 3.0-9.0 range and the heat treatment, but it was unstable with the treatment of protease (type I). The amino acid composition of the purified peaks 8-1 and 8-2 were Asx, Tyr, Gln, Pro, and Ser in a molar ratio of 3:1:1:1:1, which showed the same amino acid composition as iturin. From these results, we confirmed that antifungal compounds from B. polyfermenticus CJ6 most likely belonged to iturin A as well as surfactins and fengycins. As lipopeptides are known to act in a synergistic manner, the antifungal compounds from B. polyfermenticus CJ6 might have potential uses in biotechnology and biopharmaceutical applications.

• Microbiology and Biotechnology Letters
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• v.38 no.2
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• pp.168-176
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• 2010

Acellular dermal matrix (ADM), produced by decellularization from human cadaveric skin, has been used for various biomedical applications. A manufacturing process for ADM ($SureDerm^{TM}$) using tri-n-butyl phospahate (TnBP) and deoxycholic acids as the decellularization solution has been developed. The manufacturing process for $SureDerm^{TM}$ has 70% ethanol treatment and ethylene oxide gas sterilization for inactivating infectious microorganisms. The purpose of this study was to examine the efficacy of the 70% ethanol treatment, decellularization process using 0.1% TnBP and 2% deoxycholic acids, and EO gas sterilization process in the inactivation of viruses. A variety of experimental model viruses for human pathogens, including the human immunodeficiency virus type 1 (HIV-1), bovine herpes virus (BHV), bovine viral diarrhoea virus (BVDV), hepatitis A virus (HAV), and porcine parvovirus (PPV) were all selected for this study. Enveloped viruses such as HIV-1, BHV, and BVDV were effectively inactivated to undetectable levels by 70% ethanol treatment. However HAV and PPV showed high resistance to 70% ethanol treatment with the log reduction factors of 1.85 and 1.15, respectively. HIV-1, BHV, and BVDV were effectively inactivated to undetectable levels by decellularization process. All the viruses tested were completely inactivated to undetectable levels by EO gas treatment. The cumulative log reduction factors of HIV-1, BHV, BVDV, HAV, and PPV were $\geq12.71$, $\geq18.08$, $\geq14.92$, $\geq6.57$, and $\geq7.18$, respectively. These results indicate that the production process for $SureDerm^{TM}$ has a sufficient virus-reducing capacity to achieve a high margin of the virus safety.

• Microbiology and Biotechnology Letters
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• v.38 no.2
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• pp.177-182
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• 2010

The goal of this study was to characterize the quality and physiological functionality of some traditional rice wines in Chungnam province, Korea. Non-sterilized and commercial sterilized traditional rice wines from five traditional rice wine factories of Chungnam province were collected and investigated for nutritional components, noxious compounds and physiological functionality. Ethanol content ranged from 16.1~18.3% and pH ranged from 3.27~4.76, and they also contained 0.15% to 0.55% of total acid. All traditional rice wines contained 10.15~139.9 mg% of amino nitrogen and 2.5~25.7% of total sugar. Among organic acids, lactic acid was contained 7.4~29.6 mg%, and succinic acid and propionic acid was also contained 0.2~2.7 mg% and 0.7~8.3 mg%, respectively. Antihypertensive angiotensin I-converting enzyme inhibitory activity were showed 37.0~86.0% in all rice wines, however, fibrinolytic activity, antioxidant activity, SOD-like activity and acetylcholinesterase inhibitory activity were low or not detected.

• Microbiology and Biotechnology Letters
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• v.38 no.2
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• pp.190-197
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• 2010

To develop the safe and natural antimicrobial agents, the 68 ethanol extracts from the 61 different kinds of oriental herbal medicine were prepared and their antimicrobial activities were evaluated. The herbal medicine used were from China (46 kinds), South Korea (14 kinds), North Korea (5 kinds) and Vietnam (3 kinds), respectively, and the root (27 species) was popular part in this study. The average water content and extraction ratio for ethanol were 7.10% and 6.75%, respectively. Determination of antimicrobial activity by disc-diffusion assay at 0.5 mg/disc concentration showed that the extract of Angelica tenuissima Nakai (china), Illicium verum, Junci medulla, Rhus javanica L., Salvia miltiorrhiza Bunge and Syzygium aromaticum has strong antimicrobial activities against different food spoilage and pathogenic bacteria and fungi. Determination of MIC and MBC/MFC further showed that the extract of Syzygium aromaticum has MIC of 1.25 mg/mL and MBC/MFC of 1.25~5.00 mg/mL against Listeria monocytogenes, Bacillus subtilis, Staphylococcus epidermidis, Staphylococcus aureus, Salmonella typhimurium, Proteus vulgaris, Escherichia coli, Pseudomonas aeruginosa, Candida albicans and Saccharomyces cerevisiae. And, the extract of Junci medulla, Rhus javanica L. and Salvia miltiorrhiza Bunge showed strong antibacterial activities with MIC of 0.08~0.63 mg/mL and MBC/MFC of 0.08~2.50 mg/mL against the tested bacteria except E. coli and P. aeruginosa. In a while, the results of hemolytic activity of 68 different herbal extracts against human red blood cells showed that the extract of Angelica tenuissima Nakai has hemolytic activity at 0.5 mg/mL concentration. Therefore, Illicium verum, Junci medulla, Rhus javanica L., Salvia miltiorrhiza Bunge and Syzygium aromaticum were finally selected for natural antimicrobial resources. Further research on active substances and the mode of action of the selected herbal medicine is necessary.

• Microbiology and Biotechnology Letters
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• v.42 no.1
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• pp.93-98
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• 2014

This study was conducted to investigate the inhibitory effect of Ecklonia cava (EC) and Eisenia bicyclis (EB) ethanol extract on histamine production in mackerel. Changes in viable cell counts, histamine contents, pH and VBN of mackerel fillet treated with ethanol extracts during 25 days at 4 were measured. Treatments of EC and EB ethanol extract had reduced growth of viable cells by 2 log cycles during storage. Production of histamine was decreased by EC and EB extracts (115 and 96 ppm) when compared to the control at 5 days (384 ppm). The pH of mackerels treated with EC and EB extracts were no different, while the pH of the control increased during storage. Furthermore, the VBN of mackerels treated with EC and EB extracts were significantly decreased when compared to the control. In conclusion, EC and EB extract may reduce scombroid fish poisoning by decreasing histamine production in mackerel during refrigerated storage.

• Microbiology and Biotechnology Letters
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• v.42 no.1
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• pp.73-81
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• 2014

Salsola collina, also known as Russian thistle, is widely distributed in and around waste facilities, roadsides, and drought and semi-drought areas, and is used as a traditional folk remedy in Chinese medicine for the treatment of hypertension. In this study, we have evaluated the anti-oxidative and anti-cancer activities of the ethanol extract of S. collina Pall. (EESC), and the molecular mechanisms of its anti-cancer effects on human colon carcinoma HT29 cells. EESC exhibited anti-oxidative activity through DPPH radical scavenging capacity and showed cytotoxic activity in a dose-dependent manner in HT29 cells. After EESC treatment, HT29 cells altered their morphology, becoming smaller and irregular in shape. EESC also induced cell accumulation in the G2/M phase in a dose-dependent manner, accompanied by a decrease of cell population in the G1 phase. The G2/M arrest by EESC was associated with the increased expression of cyclin-dependent kinase (CDK) inhibitor p21 and Wee1 kinase, which phosphorylates, or inactivates, Cdc2. EESC treatment induced the phosphorylation of Cdc2 and Cdc25C, and inhibited cyclin A and Cdc25C protein expression. In addition, S arrest was induced by the highest concentration of EESC treatment, associated with a decrease of cyclin A and Cdk2 expression. These findings suggest that EESC may possess remarkable anti-oxidative activity and exert an anti-cancer effect in HT29 cells by cell cycle regulation.

• Microbiology and Biotechnology Letters
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• v.42 no.1
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• pp.58-66
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• 2014

In the course of study for a use for non-edible parts of broccoli (Brassica oleracea L), and the development of processed food utilizing these parts, edible floret and non-edible stalk were extracted with ethanol and different organic solvent fractions were prepared. With 10 different extracts and fractions, their useful components and various biological activities, such as antioxidant, antimicrobial and anti-proliferation activity, were investigated. The stalk has more abundant water soluble carbohydrate when compared with the floret, and floret has higher hexane-soluble pigments. Analysis of total flavonoid and total polyphenol contents showed that the floret has 1.5~1.99 times higher concentrations than the stalk. Among the fractions, ethylacetate (EA) fractions have the highest amount of total flavonoid and total polyphenol. The stalk and floret possessed 9.45 and 42.01 mg-total flavonoid/g, respectively. In the antioxidation activity assay, the EA fraction of floret showed strong radical scavenging activity and reducing power, while the n-hexane fraction of the stalk exhibited nitrite scavenging activity. In the antimicrobial activity assay, the EA fraction of floret showed a strong and broad-range of antibacterial activity, irrespective of gram positive or gram negative bacteria. In a while, the hexane and EA fractions revealed anti-proliferative effects against the human colorectal cancer cell HCT-116. Strong anti-proliferative activities were found in the hexane fraction of stalk (18.4% of cell viability), and the n-butanol fraction of floret (6.9% of cell viability). Our results suggest that the further study of the characterization of active fractions and the identification of active components from different parts of broccoli are needed to develop functional foods or novel plant-derived medicines.

• Microbiology and Biotechnology Letters
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• v.42 no.1
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• pp.32-40
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• 2014

Methane oxidation and the production potentials of ground soil (soil A) and garden soil (soil B, C, & D) in an urban school were evaluated, and the methanotrophic and methanogen communities in the soil samples were quantified using quantitative realtime PCR. The methanotrophic community in the raw soil A sample possessed a $6.1{\times}10^3$ gene copy number/g dry weight soil, whereas those in the raw soils B~D samples were $1.6-1.9{\times}10^5$ gene copy numbers/g dry weight soil. Serum bottles added with the soil samples were enriched with methane gas, and then evaluated for their methane oxidation potential. The soil A sample had a longer induction phase for methane oxidation than the other soils. However, soil A showed a similar methane oxidation potential with soils B~D after the induction phase. The methanotrophic community in the enriched soil A sample was increased by up to $2.3{\times}10^7$ gene copy numbers/g dry weight soil, which had no significantly difference compared with those in soils B~D ($1.2-2.8{\times}10^8$ gene copy numbers/g dry weight soil). Methane production showed a similar tendency to methane oxidation. The methanogens community in raw soil A ($1.7{\times}10^5$ gene copy number/g dry weight soil) was much less than those in raw soils B~D ($1.3-3.4{\times}10^7$ gene copy numbers/g dry weight soil). However, after methane gas was produced by adding starch to the soils, soil samples A~D showed $10^7$ gene copy numbers/g dry weight soil in methanogens communities. The results indicate that methanotrophic and methanogenic bacteria have coexisted in this urban school's soils. Moreover, under appropriate conditions for methane oxidation and production, methanotrophic bacteria and methanogens are increased and they have the potential for methane oxidation and production.

• Microbiology and Biotechnology Letters
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• v.42 no.2
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• pp.170-176
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• 2014

In this study, the anti-oxidative and anti-inflammatory activities of Euptelea pleiosperma ethanol extract (EPEE) were evaluated using in vitro assays and cell culture model systems. EPEE possessed a more potent scavenging activity against 1,1-diphenyl-2-picryl hydrazyl than the ascorbic acid used as a positive control. EPEE effectively suppressed lipopolysaccharide (LPS), in addition to hydrogen peroxide induced reactive oxygen species on RAW 264.7 cells. Furthermore, EPEE induced the expression of the anti-oxidative enzyme heme oxygenase 1 (HO-1) and its upstream transcription factor, nuclear factor-E2-related factor 2 (Nrf2), dose and time dependently. The modulation of HO-1 and Nrf2 expression might be regulated by mitogen-activated protein kinases and phosphatidyl inositol 3 kinase/Akt as their upstream signaling pathways. On the other hand, EPEE inhibited LPS induced nitric oxide (NO) formation without cytotoxicity. Suppression of NO formation was the result of the down regulation of inducible NO synthase (iNOS) by EPEE. Suppression of NO and iNOS by EPEE may be modulated by their upstream transcription factor, nuclear factor ${\kappa}B$, and AP-1 pathways. Taken together, these results provide important new insights into E. pleiosperma, namely that it possesses anti-oxidative and anti-inflammatory activities, indicating that it could be utilized as a promising material in the field of nutraceuticals.

• Microbiology and Biotechnology Letters
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• v.42 no.2
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• pp.162-169
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• 2014

In the course of study for the development of functional food using red beans (azuki beans, Phaseolus radiatus L.), the ethanol extracts from raw-red bean (RRB) and boiled-red bean (BRB) were prepared, and the components and various biological activities of both were compared. It was observed that the extraction yield, and the total polyphenol content, of the BRB were 1.2 times higher than that of the RRB. However, the contents of total flavonoid, total sugar and reducing sugar in the BRB were 30, 27.9 and 30.8% respectively when compared with those of RRB. In relation to antioxidative activity, both RRB and BRB exhibited moderate DPPH anion, ABTS cation, and nitrite scavenging activities and reducing power, though in all cases RRB demonstrated stronger activities than BRB. The extracts of RRB and BRB did not reveal any antimicrobial activities. In a ${\alpha}$-amylase inhibitory activity assay, RRB was higher than BRB, while BRB showed higher ${\alpha}$-glucosidase inhibitory activity than RRB. A strong and particular activity was observed in an anti-thrombosis activity assay of RRB. The extract of RRB demonstrated strong inhibitions against prothrombin and blood coagulation factors, with moderate thrombin inhibition. However, the extract of BRB did not exhibit any significant anti-thrombosis activity. Our results indicate that RRB has different, but useful biological activities, and loss or elimination of the biologically active substances in RRB occurs during the production of BRB. Therefore, to develop more functional foods from red beans, a study of suitable boiling, heating and drying processes is essential, and the efficient re-use of boiled waste water from the boiling process is necessary. These results could be applied to the further development of functional red bean beverages and sweat red bean pastes.