• International Journal of Industrial Entomology and Biomaterials
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• v.8 no.2
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• pp.195-200
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• 2004

Trypsin can be immobilized on silk fibroin fiber (SFF) by introducing several spacer arms, such as ethylene diamine (ED), bovine serum albumin (BSA) and silk sericin (SS). Direct immobilization on silk fiber (SFFGA) has low activity because of the steric hindrance between the trypsin and substrate. The introduction of spacer arms onto SFF-GA can enhance the activity of trypsin by reducing the steric hindrance. When ED is used as a spacer arm, the activity of trypsin has increased but its stability decreased due to the increased hydrophobicity of SFF. BSA and SS, as a spacer arm, have better results in both activity and stability. SFF-BSA shows some decrease in the specific activity due to improper immobilizatin. SFF-SS maintained 90% of its initial activity even after 12 hrs incubation at $50^{\circ}C$. In the case of repeated hydrolysis of silk sericin with immobilized trypsin, SFF-GA and SFF-ED lost 50% of their initial activity right after first run, whereas SFF-BSA and SFF-SS maintained 80% of their initial activities even after 5 runs. Higher operational stability is due to increased hydrophilicity of SFF by introducing hydrophilic spacer arms such as BSA and SS. The high content of serine in SS increases the hydrophilicity of SFF resulting the best results among other spacer arms.

• Korean Journal of Food Science and Technology
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• v.39 no.5
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• pp.564-568
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• 2007

In this study, the ethanolic extracts of 40 species of traditional herbal medicines were examined for their antimicrobial activities against Clostridium difficile. Among the 43 screened traditional herbal medicines, Achyranthes Japonica Nakai (AJN), Siegesbeckia glabrescens Makino, and Phelloedendron amurense Ruprecht showed antimicrobial activities greater than 90% at a concentration of 500 ppm. According to the minimum inhibitory concentration (MIC) test the ethyl acetate soluble fraction of the AJN ethanolic extracts (AJNEA) showed the highest growth inhibitory activity against C. difficile, with a MIC of $625{\mu}g/mL$. In addition, the effect of AJNEA on the growth of lactic acid bacteria was investigated. AJNEA did not inhibit the growth of the tested Bifidobacterium spp. or Lactobacillus spp., with the exception of B. longum, Streptococcus thermophilus, and L. helveticus. These results indicate that AJNEA can be utilized as a potential antimicrobial agent against C. difficile related disease.

• Toxicological Research
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• v.3 no.1
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• pp.1-8
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• 1987

In vitro induction of cytochrome 450 associated monooxygenase activities by phenobarbital (PB) and 3-methylcholanthrene (MC) was investigated in primary cultures of adult rat hepatocytes. PB and MC were added to the culture 24 hr after the initial plating of hepatocytes. A signiftcant increase of the activities of 7-ethoxycoumarin 0-deethylase and aryl hydrocarbon hydroxylase were observed in MC and PB treated culture. MC caused about 500% induction of the initial oxidation rates of both enzymes in 48 hr. However the PB maintained both enzyme activities close to the level of freshly isolated hepatocytes. Biphenyl 4-hydroxylase and aminopyrine N-demethylase activities were also induced by MC and PB. But the level of induction was less than that occuring with 7-ethoxycoumarin 0-deethylase and aryl hydrocarbon hydroxylase. When aflatoxin $B_1$ was added to the hepatocyte cultures which have been treated with MC or PB, it caused a significant increase of the unscheduled DNA synthesis at higher dose of aflatoxin $B_1$ as compared to those of untreated control hepatocyte cultures. The results suggest that microsomal enzyme activities can be selectively controlled preferably in hepatocyte cultures by the in vitro induction method. This principle may be useful for studying the metabolism and other toxicological studies.

• International Journal of Industrial Entomology and Biomaterials
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• v.9 no.1
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• pp.29-33
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• 2004

Low molecular weight silk fibroin (LMSF), which was prepared by hydrolysis of silk fibroin using high-temperature and high-pressure method, was found to inhibit the oxidation of L-3,4,-dihydroxyphenylalanine (L-DOPA) catalyzed by mushroom tyrosinase (EC 1.14.18.1). LMSF contained mostly free amino acids such as L-glycine, L-alanine, and L-serine and oligopeptides, mainly glycine-alanine dimer. As a result of analyzing the inhibition kinetics from Lineweaver-Burk plots, L-glycine and glycine-alanine dimer showed noncompetitive behavior while uncompetitive behavior was observed in L-alanine, and L-serine. When weight percent concentration of ${ID_50}$ was compared, L-glycine was most effective on the inhibition and LMSF was also good enough for the inhibition effect of tyrosinase activity. LMSF showed a mixed-type inhibition and the inhibitory mechanism of LMSF might be caused by free amino acids and oligopeptides. As a result of spectroscopic observation with time, initial rate of increase of DOPAchrome decreased remarkably and the time to reach maximum absorbance increased as an increase of the concentration of L-glycine, meaning that L-glycine made itself mainly responsible for the formation of chelate with ${Cu^2+}$ in tyrosinase. However, in case of L-alanine, L-serine, and especially glycine-alanine dimmer, the production of DOPAchrome after an arrival at maximum absorbance decreased, indicating the production of adducts through the reaction with DOPAquinone.

• Clean Technology
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• v.17 no.1
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• pp.37-40
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• 2011

Some ionic liquids are suitable for catalysts and solvents which are applicable to $CO_2$ fixation reaction converting $CO_2$ to carbonate. Using the ionic liquids, the synthesis process will become greener and simpler because of easy catalyst recycling and unnecessary use of volatile and harmful organic solvents. In this work, the synthesis of propylene carbonate from propylene oxide using carbon dioxide and ionic liquids were measured at high pressures up to ~140 bar and at temperatures between $60^{\circ}C$ and $80^{\circ}C$. As a results, we found the optimum condition and obtained the maximum yield under that condition.

• Microbiology and Biotechnology Letters
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• v.35 no.1
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• pp.52-57
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• 2007

A gas-circulating bioreactor was used for enrichment of autotrophic methanogens. Mixture of hydrogen and carbon dioxide (5:1) was used as a sole energy and carbon source. Anaerobic digestive sludge isolated from wastewater treatment system was inoculated into the gas-circulating bioreactor. The enrichment of two chemolithotrophic methanogens, Methanobacterium curvum and Methanobacterium oryzae was accomplished in the gas-circulating bioreactor. The enriched bacteria were cultivated in a bioreactor equipped with hollow-fiber hydrogen-supplying system (hollow-fiber bioreactor), and a hybrid-type bioreactor equipped with hollow-fiber hydrogen-supplying system and electrochemical redox control system. The methane productivity was maximally 30% (V/V) in the hollow-fiber bioreactors and 50% (V/V) in the hybrid-type bioreactor.

• Journal of Microbiology and Biotechnology
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• v.28 no.2
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• pp.275-283
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• 2018

Ischemic stroke can result from blockage of blood vessels, forming fibrin clots in the body and causing irreparable brain damage. Remedial thrombolytic agents or anticoagulants have been studied; however, because the FDA-approved tissue plasminogen activator has low efficacy and side effects, it is necessary to develop safer and more effective treatment candidates. This study aimed at assessing the fibrinolytic and anticoagulation features of a novel serine protease extracted and purified from Diopatra sugokai, a polychaeta that inhabits tidal flats. The purified serine protease was obtained through ammonium sulfate precipitation, affinity chromatography, and ion-exchange chromatography. Its molecular size was identified via SDS-PAGE. To characterize its enzymatic activities, the protease activity at various pH and temperatures, and in the presence of various inhibitors, was measured via azocasein assay. Its fibrinolytic activity and anticoagulant effect were assessed by fibrin zymography, fibrin plate assay, and fibrinogenolytic activity assays. The novel 38 kDa serine protease had strong indirect thrombolytic activity rather than direct activity over broad pH (4-10) and temperature ($37^{\circ}C-70^{\circ}C$) ranges. In addition, the novel serine protease exhibited anticoagulant activity by degrading the ${\alpha}$-, ${\beta}$-, and ${\gamma}$-chains of fibrinogen. In addition, it did not produce cytotoxicity in endothelial cells. Therefore, this newly isolated serine protease is worthy of further investigation as a novel alkaline serine protease for thrombolytic therapy against brain ischemia.

• Journal of Microbiology and Biotechnology
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• v.19 no.9
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• pp.1019-1027
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• 2009

A noncompartmented microbial fuel cell (NCMFC) composed of a Mn(IV)-carbon plate and a Fe(III)-carbon plate was used for electricity generation from organic wastewater without consumption of external energy. The Fe(III)-carbon plate, coated with a porous ceramic membrane and a semipermeable cellulose acetate film, was used as a cathode, which substituted for the catholyte and cathode. The Mn(IV)-carbon plate was used as an anode without a membrane or film coating. A solar cell connected to the NCMFC activated electricity generation and bacterial consumption of organic matter contained in the wastewater. More than 99% of the organic matter was biochemically oxidized during wastewater flow through the four NCMFC units. A predominant bacterium isolated from the anode surface in both the conventional and the solar cell-linked NCMFC was found to be more than 99% similar to a Mn(II)-oxidizing bacterium and Burkeholderia sp., based on 16S rDNA sequence analysis. The isolate reacted electrochemically with the Mn(IV)-modified anode and produced electricity in the NCMFC. After 90 days of incubation, a bacterial species that was enriched on the Mn(IV)-modified anode surface in all of the NCMFC units was found to be very similar to the initially isolated predominant species by comparing 16S rDNA sequences.

• Journal of Microbiology and Biotechnology
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• v.12 no.1
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• pp.176-181
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• 2002

In order to isolate and identify Bifidobacterium spp. that originated in Korea, feces were sampled from healthy Korean adults and children living in three villages, the first having a history of longevity and the other two where the diet did not include fermented milk or any pharmaceutical preparations. Through the use of Gram staining and microscopic examination for cell morphology, 23 bacterial strains presumed to be the Bifidobacterium genus were isolated from the feces of 13 out of a total of 59 Korean people. To identify the Bifidobacterium strains at the genus level, these bacteria were then analyzed by TLC and the fructose-6-phosphate phosphoketolase (F6PPK) test. The result showed that 22 of the isolated strains were confirmed to be members of the genus Bifidobacterium. All of these bifidobacteria were also identified as Bifidobacterium spp. by the fermentation test. Using a RFLP analysis, an attempt was made to identify the Bifidobacterium spp. that had been isolated from both Korean adults and children. In a genomic Southern blot analysis after digestion with two restriction enzymes (EcoRI, HindIII), all of the 14 randomly selected Korean isolates showed patterns identical to those of three different B. longum species. Another restriction enzyme, CfoI (4-bp recognition enzyme), was then used to identify the strain. Interestingly, all the Korean isolates were identified as B. longum ATCC 15708, indicating that a RFLP analysis was effective for identifying Bifidobacterium spp. at both the strain and species levels.

• Journal of Microbiology and Biotechnology
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• v.20 no.2
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• pp.356-362
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• 2010

Ceramide is important not only for the maintenance of the barrier function of the skin but also for the water-binding capacity of the stratum corneum. Although the exact role of ceramide in the human skin is not fully understood, ceramide has become a widely used ingredient in cosmetic and pharmaceutical industries. Compared with other microorganisms, yeast is more suitable for the production of ceramide because yeast grows fast and is non-toxic. However, production of ceramide from yeast has not been widely studied and most work in this area has been carried out using Saccharomyces cerevisiae. Regulating the genes that are involved in sphingolipid synthesis is necessary to increase ceramide production. In this study, we investigated the effect of the genes involved in the synthesis of ceramide, lcb1, lcb2, tsc10, lac1, lag1, and sur2, on ceramide production levels. The genes were cloned into pYES2 high copy number vectors. S. cerevisiae was cultivated on YPDG medium at $30^{\circ}C$. Ceramide was purified from the cell extracts by solvent extraction and the ceramide content was analyzed by HPLC using ELSD. The maximum production of ceramide (9.8 mg ceramide/g cell) was obtained when the tsc10 gene was amplified by the pYES2 vector. Real-time RT-PCR analysis showed that the increase in ceramide content was proportional to the increase in the tsc10 gene expression level, which was 4.56 times higher than that of the control strain.