• Journal of Microbiology and Biotechnology
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• v.22 no.10
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• pp.1395-1400
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• 2012

In this study, we investigated various cultural and operational factors to enhance electricity generation in a microbial fuel cell (MFC) using Geobacter sulfurreducens. The pure culture of G. sulfurreducens was cultivated using various substrates including acetate, malate, succinate, and butyrate, with fumarate as an electron acceptor. Cell growth was observed only in acetate-fed medium, when the cell concentrations increased 4-fold for 3 days. A high acetate concentration suppressed electricity generation. As the acetate concentration was increased from 5 to 20 mM, the power density dropped from 16 to $13mW/m^2$, whereas the coulombic efficiency (CE) declined by about half. The immobilization of G. sulfurreducens on the anode considerably reduced the enrichment period from 15 to 7 days. Using argon gas to create an anaerobic condition in the anode chamber led to increased pH, and electricity generation subsequently dropped. When the plain carbon paper cathode was replaced by Pt-coated carbon paper (0.5 mg $Pt/cm^2$), the CE increased greatly from 39% to 83%.

• Microbiology and Biotechnology Letters
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• v.48 no.4
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• pp.491-505
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• 2020

A newly isolated strain, Proteus vulgaris OR34, from olive mill waste was found to secrete an alkaline extracellular lipase at 11 U·ml-1 when cultivated on an optimized liquid medium. This lipase was purified 94.64-fold with a total yield of 9.11% and its maximal specific activity was shown to be 3232.58 and 1777.92 U·mg-1 when evaluated using the pH-stat technique at 55℃ and pH 9 and Tributyrin TC4 or olive oil as the substrate. The molecular mass of the pure OR34 lipase was estimated to be around 31 kDa, as revealed by SDS-PAGE and its substrate specificity was investigated using a variety of triglycerides. This assay revealed that OR34 lipase preferred short and medium chain fatty acids. In addition, this lipase was stable in the presence of high concentrations of bile salt (NaDC) and calcium ions appear not to be necessary for its activity. This lipase was inhibited by THL (Orlistat) which confirmed its identity as a serine enzyme. In addition, the immobilization of OR34 lipase by adsorption onto calcium carbonate increased its stability at higher temperatures and within a larger pH range. The immobilized lipase exhibited a high tolerance to organic solvents and retained 60% of its activity after 10 months of storage at 4℃. Finally, the OR34 lipase was applied in biodiesel synthesis via oleic acid mediated esterification of methanol when using hexane as solvent. The best conversion yield (67%) was obtained at 12 h and 40℃ using the immobilized enzyme and this enzyme could be reused for six cycles with the same efficiency.

• Applied Chemistry for Engineering
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• v.10 no.6
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• pp.852-856
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• 1999

The objective of this study was to prepare hydrogel beads which were useful microbial immobilization to remove nitrogen and phosphorous in the industrial wastewater. Two different polyols(PEG, PTMG) terminated with photo-crosslinkable methacrylate groups were synthesized. Structures of the prepolymers and the UV cured hydrogels were characterized by using $^1H$-NMR and FT-IR spectroscopy. Water content, mechanical strength and pore sizes of the hydrogels having different MW of polyols and different ratios of PEG/PTMG were investigated. Hydrogels prepared from PEG(MW1000) only or the mixture of PEG(MW1000) and PTMG(MW2900) with 7:3 by weight were considered as potential candidates for the matrix for the immobilization of microorganism.

• KSBB Journal
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• v.17 no.3
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• pp.213-227
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• 2002

To date, the majority of biosensor technologies use binding components such as enzymes antibodies, nucleic acids and protein ligands. In contrast, the goal underlying the use of cells and tissues of animals and plants for a sensor system is to obtain systems capable of extracting information based on the biological activity, mechanisms of action and consequences of exposure to a chemical or biological agent of interest. These systems enable the interrogation of more complex biological response and offer the potential to gather higher information content from measuring physiologic and metabolic response. In these articles, same of the recent trends and applications of microbial biosensors in environmental monitoring and for use in food and fermentations have been reviewed. This endeavor presents many technological challenges to fabricate new microbial biosensors for other scientific field.

• Journal of Microbiology
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• v.44 no.3
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• pp.354-359
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• 2006

This study examined the capacity of immobilized bacteria to degrade petroleum hydrocarbons. A mixture of hydrocarbon-degrading bacterial strains was immobilized in alginate and incubated in crude oil-contaminated artificial seawater (ASW). Analysis of hydrocarbon residues following a 30-day incubation period demonstrated that the biodegradation capacity of the microorganisms was not compromised by the immobilization. Removal of n-alkanes was similar in immobilized cells and control cells. To test reusability, the immobilized bacteria were incubated for sequential increments of 30 days. No decline in biodegradation capacity of the immobilized consortium of bacterial cells was noted over its repeated use. We conclude that immobilized hydrocarbon-degrading bacteria represent a promising application in the bioremediation of hydrocarbon-contaminated areas.

• Journal of Microbiology and Biotechnology
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• v.24 no.5
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• pp.639-647
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• 2014

Laccases have a broad range of industrial applications. In this study, we immobilized laccase on $SiO_2$ nanoparticles to overcome problems associated with stability and reusability of the free enzyme. Among different reagents used to functionally activate the nanoparticles, glutaraldehyde was found to be the most effective for immobilization. Optimization of the immobilization pH, temperature, enzyme loading, and incubation period led to a maximum immobilization yield of 75.8% and an immobilization efficiency of 92.9%. The optimum pH and temperature for immobilized laccase were 3.5 and $45^{\circ}C$, respectively, which differed from the values of pH 3.0 and $40^{\circ}C$ obtained for the free enzyme. Immobilized laccase retained high residual activities over a broad range of pH and temperature. The kinetic parameter $V_{max}$ was slightly reduced from 1,890 to 1,630 ${\mu}mol/min/mg$ protein, and $K_m$ was increased from 29.3 to 45.6. The thermal stability of immobilized laccase was significantly higher than that of the free enzyme, with a half-life 11- and 18-fold higher at temperatures of $50^{\circ}C$ and $60^{\circ}C$, respectively. In addition, residual activity was 82.6% after 10 cycles of use. Thus, laccase immobilized on $SiO_2$ nanoparticles functionally activated with glutaraldehyde has broad pH and temperature ranges, thermostability, and high reusability compared with the free enzyme. It constitutes a notably efficient system for biotechnological applications.

• Journal of Sensor Science and Technology
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• v.23 no.1
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• pp.35-41
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• 2014

Microbes have been used extensively in various fields of researches and industries but has not been used widely for microfluidic biosensor applications because it is difficult to immobilize properly to a small space. Therefore, we developed a microbial immobilization method for microfluidic devices using single-walled nanotubes and dielectrophoretic force. Single-walled nanotubes and Escherichia coli were aligned between two cantilever electrodes by a positive dielectrophoretic force resulting in a film of single-walled nanotubes with attached Escherichia coli. The optimal condition of film formation without a cell lysis was investigated. Diameter of single-walled nanotubes and electric field (intensity and duration of application) had an effect on the cell viability. On the other hand, the cell concentration of the suspension did not affect the cell viability. Paraoxon was detected using single-walled nanotubes film with attached Escherichia coli that expressed organophosphorus hydrolase. This film which is suspended from the substrate showed faster response time than sensors that are not suspended from the substrate.

• Korean Journal of Food Science and Technology
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• v.30 no.5
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• pp.1222-1228
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• 1998

Stress will induce various changes in human metabolism. The remarkable phenomenon of these changes is increased energy metabolism that can induce many reactive oxygen species (ROS) production. ROS can peroxidize cellular macromolecules including lipid and protein. The object of this study was to investigate that stress may induce cellular damage by producing ROS and that Rooibos tea can protect cells against reactive oxygen species by immobilization stress in SD rat. The stress group significantly increased in 5-hydroxyindole acetic acid (5-HIAA), one of the stress hormone. Rooibos tea treatment had no effects on 5-HIAA contents, but body weight of Rooibos tea treated rat more increased than that of only the stress group. It was suggested that Rooibos tea colud not affect stress response itself, but protect against the another mechanism. We thought that the oxidative damage was caused by increased energy metabolism. Protein degradation level and lipid peroxide formation on index of oxidative damage significantly increased in the stress group. But the stress-induced activity change could not be observed in antioxidative enzymes such as superoxide dismutase, glutathione peroxidase and glutathione reductase. But the catalase activity of the brain significantly was inhibited by the stress. From these results, it was suggested that the immobilization stress induce the brain oxidative damage. However the oxidative damage was inhibited by feeding Rooibos tea containing various antioxidants, such as polyphenol, flavonoid and so on. Therefore, Rooibos tea have the protective effects against the stress caused by the ROS mediated cellular damage.

• Journal of Microbiology and Biotechnology
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• v.22 no.6
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• pp.818-825
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• 2012

Keratinases are exciting keratin-degrading enzymes; however, there have been relatively few studies on their immobilization. A keratinolytic protease from Chryseobacterium sp. kr6 was purified and its partial sequence determined using mass spectrometry. No significant homology to other microbial peptides in the NCBI database was observed. Certain parameters for immobilization of the purified keratinase on chitosan beads were investigated. The production of the chitosan beads was optimized using factorial design and surface response techniques. The optimum chitosan bead production for protease immobilization was a 20 g/l chitosan solution in acetic acid [1.5% (v/v)], glutaraldehyde ranging from 34 g to 56 g/l, and an activation time between 6 and 10 h. Under these conditions, above 80% of the enzyme was immobilized on the support. The behavior of the keratinase loading on the chitosan beads surface was well described using the Langmuir model. The maximum capacity of the support ($q_m$) and dissociation constant ($K_d$) were estimated as 58.8 U/g and 0.245 U/ml, respectively. The thermal stability of the immobilized enzyme was also improved around 2-fold, when compared with that of the free enzyme, after 30 min at $65^{\circ}C$. In addition, the activity of the immobilized enzyme remained at 63.4% after it was reused five times. Thus, the immobilized enzyme exhibited an improved thermal stability and remained active after several uses.

• Korean Journal of Food Science and Technology
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• v.16 no.3
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• pp.267-272
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• 1984

${\beta}-Fructofuranosidase$ was immobilized covalently on the oxidized microbial wall of a Penicillium spp. 'PS-8', which is totally different from the conventional whole cell immobilization in concept. The immobilization of ${\beta}-fructofuranosidase$ by a series of treatments; oxidation of microbial cells with sodium metaperiodate, enzyme loading on the oxidized cells, extrusion, and crosslinking induced by glutaradehyde, were carried out. The final product had a good mechanical strength and showed 26% of the applied enzyme activity. The specific activity was 750 units per g of the dry cell product. The immobilized enzyme showed the kinetic parameters as follows; optimum pH at 5, optimum temperature at $55^{\circ}C$, activation energy of 19 kJ $mol^{-1}$, and apparent Km of 55 mM.