• Biotechnology and Bioprocess Engineering:BBE
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• 제2권2호
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• pp.141-146
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• 1997

A new sensing system based on the immobilization of luminescent batcteria, Photobacterium phosphoreum, was proposed for continuous real-time monitoring of polluants. The response curves demonstrate that Photobacterium phosphoreum immobilized on the strontium alginate was very sensitive to seven reference chemicals used. The significant inhibitory concentrations for bioluminescence emission were 5 ppm for Pb(NO3)2, NiCl2, CdCl2, 50 ppm for NaAsO2, 0.1ppm for HgCl2, 0.5ppm for pentachlorophenol and less than 5ppm for SDS, respectively. The alginate mixed-cells (AMC) retained their luminescence during experimental period (29 days) under storage condition of -8$0^{\circ}C$. The variables affecting performance of continuous flow through monitoring (CFTM) were optimized in order to ensure stability and efficiency. The flow through cell with strontium-alginate immobilized luminescent bacteria was tested with salicylate and 4-nitrophenol and a rapid response of luminescence was recorded by time drive mode in bioluminescence spectrometer after exposure to both toxicants.

• Journal of Microbiology and Biotechnology
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• 제17권9호
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• pp.1533-1537
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• 2007

In this study, in order to develop a continuous production process of lactosucrose in a packed-bed reactor, Sterigmatomyces elviae ATCC 18894 was selected and mutated. The mutant strain of S. elviae showed 54.3% higher lactosucrose production than the wild type. Reaction conditions such as temperature, pH, substrate concentration and flow rate were also optimized. Under optimized reaction conditions ($50^{\circ}C$, pH 6.0, 25% sucrose and 25% lactose as substrate, flow rate 1.2 ml/min), the maximum concentration of lactosucrose (192 g/l) was obtained. In a packed-bed reactor, continuous production of lactosucrose was performed using S. elviae mutant immobilized in calcium alginate, and about 180 g/l of lactosucrose production was achieved for 48 days.

• Journal of fish pathology
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• 제27권2호
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• pp.99-105
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• 2014

Biosensors consist of biochemical recognition agents like antibodies immobilized on the surfaces of transducers that change the recognition into a measurable electronic signal. Here we report a piezoelectric immunosensor made to detect Vibrio vulnificus. A 9MHz AT-cut piezoelectric wafer attached with two gold electrodes of 5mm diameter was used as the transducer of the QCM biosensor with a reproducibility of ${\pm}0.1Hz$ in frequency response. We have tried different approaches to immobilize antibody on the sensor chip. Concerning the orientation of antibody for the best antigen binding capacity, the antibody was immobilized by specific binding to protein G or by cross-linking through hydrazine. In addition, protein G was cross-linked on glutaraldehyde activated immine layer (PEI) or EDC/NHS activated sulfide monolayer (MPA). PEI was found to be more effective to immobilize protein G following glutaraldehyde activation than MPA. However, hydrazine chip showed a better capability to immobilize more IgG than protein G chip and a higher sensitivity. The sensor system was able to detect V. vulnificus in dose dependent manner and was able to detect bacterial cells within 5 minutes by monitoring frequency shifts in real time. The detection limit can be improved by preincubation to enrich the bacterial cell number.

• KSBB Journal
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• 제11권6호
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• pp.649-653
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• 1996

Imnmobilization characteristics of Thiobacillus sp. IW which oxidizes sulfur compound was studied to use the bacterium in odor controlling equipment for the future. The optimum growth conditions for Thiobacillus sp. IW were pH7, $30^{\circ}C$ and the generation time was 38min, which was extremely fast compared with other sulfur oxidizing bacteria. Optimum growth conditions in activated carbon as a carrier was pH5, $35^{\circ}C$ and those in bioceramics was pH 7∼8, $35^{\circ}C$. Cell growth immobilized in bioceramics was more stable in pH, temperature change than that immobilized in activated carbon and total number of cells in bioceramics were also higher. Based on these results, the bioceramics is thought to be better carrier in immobilization of Thiobacillus sp. IW.

• Journal of Microbiology and Biotechnology
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• 제26권8호
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• pp.1348-1357
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• 2016

An enzymatic reaction system was developed and optimized for bioconversion of resveratrol from glucose. Liquid enzyme extracts were prepared from Alternaria sp. MG1, an endophytic fungus from grape, and used directly or after immobilization with sodium alginate. When the enzyme solution was used, efficient production of resveratrol was found within 120 min in a manner that was pH-, reaction time-, enzyme amount-, substrate type-, and substrate concentration-dependent. After the optimization experiments using the response surface methodology, the highest value of resveratrol production (224.40 μg/l) was found under the conditions of pH 6.84, 0.35 g/l glucose, 0.02 mg/l coenzyme A, and 0.02 mg/l ATP. Immobilized enzyme extracts could keep high production of resveratrol during recycling use for two to five times. The developed system indicated a potential approach to resveratrol biosynthesis independent of plants and fungal cell growth, and provided a possible way to produce resveratrol within 2 h, the shortest period needed for biosynthesis of resveratrol so far.

• Natural Product Sciences
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• 제13권2호
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• pp.128-131
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• 2007

Tumor angiogenesis is a critical step f3r the growth and metastasis of solid tumors. Vascular endothelial growth factor (VEGF) is the most important angiogenic molecule associated with tumor-induced neovascularization. VEGF exerts its activity through binding to its receptor tyrosine kinase, KDR/Flk-1, expressed on the surface of endothelial cells. This study was carried out to investigate inhibitory effect of extracts from root cortex of Paeonia suffruticosa Andrews on VEGF binding to VEGF receptor. The MeOH extract from P. suffrutiocosa Andr. inhibited the binding of KDR/Flk-1-Fc to immobilized VEGF$_{165}$ more than 45% at the concentration of 100 ${\mu}$g/mL. The MeOH extract was further fractionated into n-hexane, ethyl acetate, n-BuOH, and aqueous fractions. Among the four fractions, the ethyl acetate fraction from the root cortex of P. suffruticosa Andr. exhibited highly effective inhibition (${\approx}$ 79% inhibition) and then n-BuOH fraction (${\approx}$ 45% inhibition) on the binding of KDR/Flk-1-Fc to immobilized VEGF$_{165}$ at the concentration of 100 ${\mu}$g/mL. The ethyl acetate fraction from the root cortex of P. suffruticosa Andr. more efficiently blocked VEGF-induced human umbilical vein endothelial cell proliferation, than the growth of HT1080 human fibrosarcoma. Our results suggest that P. suffruticosa Andr. may be used as a candidate fur developing anti-angiogenic agent.

• Journal of the Korean Society of Food Science and Nutrition
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• 제24권5호
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• pp.803-810
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• 1995

Co-immobilized mixed culture system(A-Z system) composed of two different oxygen-demanding strains, aerobic(Aspergillus awamori) and anaerobic(Zymomonas mobilis) strains, in a Ca-alginate gel beads was developed to increase ethanol production from raw starch as a carbon source. Optimal mixture ratio of A. awamori and Z. mobilis was $1.25{\times}10^{9}\;spores/L-gel$ and 0.5g cells/L-gel, respectively. After 120 hours of cultivation, gel beads distinguished oxygen-rich surface for A. awamori from oxygen-deficient central part for Z. mobilis. At A-Z culture system, yield of ethanol on glucose, $Y_{p/s}=0.18$, was very low and there was high leakage of cells from surface of gel beads. At A-Z 36 cultrue system with changing silicon check valve for cotton plug at 36 hours in A-Z culture system, there was no cell leakage from gel beads, pH was maintained at around 4.3 during cultivation, and yield of ethanol on glucose, $Y_{p/s}=0.36$, showed 2 times higher than that of control culture system(cotton plug culture).

• Journal of Microbiology and Biotechnology
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• 제27권3호
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• pp.507-513
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• 2017

Bacillus subtilis spores can be used for protein display to engineer protein properties. This method overcomes viability and protein-folding concerns associated with traditional protein display methods. Spores remain viable under extreme conditions and the genotype/phenotype connection remains intact. In addition, the natural sporulation process eliminates protein-folding concerns that are coupled to the target protein traveling through cell membranes. Furthermore, ATP-dependent chaperones are present to assist in protein folding. CotA was optimized as a whole-cell biocatalyst immobilized in an inert matrix of the spore. In general, proteins that are immobilized have advantages in biocatalysis. For example, the protein can be easily removed from the reaction and it is more stable. The aim is to improve the pH stability using spore display. The maximum activity of CotA is between pH 4 and 5 for the substrate ABTS (ABTS = diammonium 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate). However, the activity dramatically decreases at pH 4. The activity is not significantly altered at pH 5. A library of approximately 3,000 clones was screened. A E498G variant was identified to have a half-life of inactivation ($t_{1/2}$) at pH 4 that was 24.8 times greater compared with wt-CotA. In a previous investigation, a CotA library was screened for organic solvent resistance and a T480A mutant was found. Consequently, T480A/E498G-CotA was constructed and the $t_{1/2}$ was 62.1 times greater than wt-CotA. Finally, E498G-CotA and T480A/E498G-CotA yielded 3.7- and 5.3-fold more product than did wt-CotA after recycling the biocatalyst seven times over 42 h.

• KSBB Journal
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• 제11권5호
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• pp.613-621
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• 1996

The effects of important medium components such as carbon, nitrogen sources and amino acids on the production of cyclosporin A(CyA) were investigated in an immobilized fungal cell fermentation using Tolypocladium inflatum. As carbon sources in the synthetic medium, fructose and maltose stimulated CyA production remarkably compared to glucose when ammonium sulfate was supplemented as a nitrogen source. In the absence of ammonium sulfate in the medium, however, CyA biosynthesis was reduced considerably without regard to C-sources tested. Ammonium sulfate was found to be the best N-source, and also ammonium phosphate and ammonium citrate showed some positive effects on CyA production. Optimum concentration of ammonium sulfate was 10g/L, and supplementation of ammonium sulfate at the start of fermentation was found to be the most efficacious for maximal production of CyA. Among the constituent amino acids of cyclic peptide, CyA, L-valine had the most significant effect on the biosynthesis of CyA, and maximum CyA production was observed when 10 g/L of L-valine was initially added.

• Journal of Microbiology and Biotechnology
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• 제15권1호
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• pp.80-85
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• 2005

Electrochemical control of the metabolic flux of W. kimchii sk10 on glucose and pyruvate was studied. The growing cell of W. kimchii sk10 produced 87.4 mM lactate, 69.3 mM ethanol, and 4.9mM lactate from 83.1mM glucose under oxidation condition of the anode compartment, but 98.9 mM lactate, 84.3mM ethanol, and 0.2 mM acetate were produced from 90.8 mM glucose under reduction condition of the cathode compartment for 24 h, respectively. The resting cell of W. kimchii sk10 produced 15.9 mM lactate and 15.2 mM acetate from 32.1 mM pyruvate under oxidation condition of the anode compartment, and 71.3 mM lactate and 3.8 mM acetate from 79.8mM pyruvate under reduction condition of the cathode compartment. The redox balance (NADH/$NAD^+$) of metabolites electrochemically produced from pyruvate was 1.05 and 18.76 under oxidation and reduction conditions, respectively. On the basis of these results, we suggest that the neutral red (NR) immobilized in bacterial membrane can function as an electron channel for the electron transfer between electrode and cytoplasm without dissipation of membrane potential, and that the bacterial fermentation of W. kimchii sk10 can be shifted to oxidized or reduced pathways by the electrochemical oxidation or reduction, respectively.