• Microbiology and Biotechnology Letters
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• v.44 no.4
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• pp.504-511
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• 2016

Lactulose, a synthetic disaccharide, has received increasing interest because of its role as a prebiotic that can increase the proliferation of Bifidobacterium and Lactobacillus spp. and enhance the absorption of calcium and magnesium. While the industrial production of lactulose is still mainly achieved by the chemical isomerization of lactose in alkaline media, this process has drawbacks including the need to remove catalysts and by-products, as well as high energy requirements. Recently, the use of cellobiose 2-epimerase (CE) has been considered an interesting alternative for industrial lactulose production. In this study, to develop a process for enzymatic lactulose production using CE, we screened improved mutant enzymes ($CS-H^RC^E$) from a library generated by an error-prone PCR technique. The thermostability of one mutant was enhanced, conferring stability up to $75^{\circ}C$, and its lactulose conversion yield was increased by 1.3-fold compared with that of wild-type CE. Using a recombinant Escherichia coli strain harboring a CS35 $H^RC^E$-expressing plasmid, we prepared cell beads immobilized on a Ca-alginate substrate and optimized their reaction conditions. In a batch reaction with 200 g/l lactose solution and the immobilized cell beads, lactose was converted into lactulose with a conversion yield of 43% in 2 h. In a repeated 38-plex batch reaction, the immobilized cell beads were relatively stable, and 80% of the original enzyme activity was retained after 4 cycles. In conclusion, we developed a reasonable method for lactulose production by immobilizing cells expressing thermostable CE. Further development is required to apply this approach at an industrial scale.

• Korean Journal of Soil Science and Fertilizer
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• v.39 no.3
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• pp.157-162
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• 2006

Objective of this research was to evaluate optimal conditions of arsenic adsorption in water by zero-valent iron (ZVI). Batch experiment showed that adsorption of arsenic by ZVI followed a Langmuir isotherm model. The masses of As(V) adsorbed onto ZVI were increased as decreasing pH of the reacting solution (pH 3: 2.05, pH 5: 1.82, pH 7: 1.24, pH 9: 1.03 mg As/g $Fe^0$) and as increasing the temperature ($15^{\circ}C$ : 1.59, $25^{\circ}C$ : 1.81, 35 : $1.93^{\circ}C$ mg As/g $Fe^0$). The SEM and EDS (energy dispersive X-ray spectrometer) analysis of morphology and structure of ZVI before and after reacting with arsenic in water revealed that a relatively smooth and large surface of ZVI was transformed into a coarse and small surface particle after the reaction. The EDS spectra on the chemical composition of ZVI demonstrated that arsenic was incorporated into ZVI by adsorption mechanism. The XRD analysis also identified that the only peak for $Fe^0$ in the ZVI before the reaction and confirmed that $Fe^0$ was transformed into $Fe_2O_3$ and FeOOH, and As into $FeAsO_4{\cdot}2H_2O$.

• Journal of Life Science
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• v.27 no.11
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• pp.1381-1392
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• 2017

This review described solvent-tolerant lipases and their potential industrial, biotechnological and environmental impacts. Although organic solvent-tolerant lipase was first reported in organic solvent-tolerant bacterium, many organic solvent-tolerant lipases are in not only solvent-tolerant bacteria but also solvent-intolerant bacterial and fungal strains, such as the well-known Bacillus, Pseudomonas, Streptomyces and Aspergillus strains. As these lipases are not easily inactivated in organic solvents, there is no need to immobilize them in order to prevent an enzyme inactivation by solvents. Therefore, the solvent-tolerant lipases have the potential to be used in many biotechnological and biotransformation processes. With the solvent-tolerant lipases, a large number insoluble substrates become soluble, various chemical reactions that are initially impossible in water systems become practical, synthesis reactions (instead of hydrolysis) are possible, side reactions caused by water are suppressed, and the possibility of chemoselective, regioselective and enantioselective transformations in solvent and non-aqueous systems is increased. Furthermore, the recovery and reuse of enzymes is possible without immobilization, and the stabilities of the lipases improve in solvent and non-aqueous systems. Therefore, lipases with organic-solvent tolerances have attracted much attention in regards to applying them as biocatalysts to biotransformation processes using solvent and non-aqueous systems.

• Korean Journal of Food Science and Technology
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• v.30 no.1
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• pp.22-34
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• 1998

The purpose of this study is to develop biosensor for determination of glucose, lactate, and ethanol in foods and food-stuffs simultaneously. The multiple cathode system was prepared with an oxygen electrode having one anode and hexagonal cathode. Glucose oxidase, mutarotase, lactate oxidase, alcohol oxidase and catalase were used for immobilization to determine glucose, lactate, and ethanol. These components including ethanol were simultaneously determined by the immobilized enzymes in the multiple cathode system. The determination of the components by enzyme sensor was based on the maximum slope of oxygen consumption from enzyme reaction of each sensor part. The response time for analysis was 1 min. The optimum condition for glucose, lactate and ethanol sensor was found to be 0.1 M potassium phosphate buffer, pH 7.0 at $40^{\circ}C$. Interferences of various sugars and organic acids were investigated. Less than 10% of error was found in determination of the components except organic acids. This difference was compensated by the modified equation. This system was confirmed by conventional methods. It was concluded that the multiple cathode system of this study is for an effective method to determine sugar, organic acid, ethanol simultaneously in foods.

• Journal of Life Science
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• v.25 no.3
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• pp.345-348
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• 2015

An organic solvent stable lipase from solvent-tolerant Pseudomonas sp. BCNU 171 had an optimal pH of 8 and an optimal temperature of 37℃. This crude extracellular lipase from BCNU 171 exhibited increased stability in the presence of various types of solvents at high concentrations (25%, v/v). The lipase stability was found to be highest in the presence of xylene (137%), followed by toluene (131%), octane (130%), and butanol (104%). Overall, BCNU 171 lipase tended to be more stable than immobilized commercial lipase (Novozyme435) in the presence of organic solvents. Furthermore, BCNU 171 lipase maintained about 90% of its enzyme original activity in the presence of NH₄⁺, Na⁺, Ba²⁺, Hg²⁺, Ni²⁺, Cu²⁺, and Ca²⁺ion and significantly increased its enzyme activity in the presence of various emulsifying agents. Thus, the organic solvent stable lipase from Pseudomonas sp. BCNU 171 could be usable as a potential whole cell biocatalyst and for synthetic applications of enzymes for industrial chemical processes in organic solvents without using immobilization.

• Journal of Life Science
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• v.23 no.12
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• pp.1471-1476
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• 2013

The present study deals with the immobilization of Kluyveromyces lactis ${\beta}$-galactosidase on a weak ionic exchange resin (Duolite A568) as polymer support. ${\beta}$-Galactosidase was immobilized using the adsorption method. A kinetic study of the immobilized enzyme was performed in a packed-bed reactor. The adsorption of the enzyme followed a typical Freundlich adsorption isotherm. The adsorption parameters of k and n were 14.6 and 1.74, respectively. The initial rates method was used to characterize the kinetic parameters of the free and immobilized enzymes. The Michaelis-Menten constant ($K_m$) for the immobilized enzyme (120 mM) was higher than it was for the free enzyme (79 mM). The effect of competitive inhibition kinetics was studied by changing the concentration of galactose in a recycling packed-bed reactor. The kinetic model with competitive inhibition by galactose was best fitted to the experimental results with $V_m$, $K_m$, and $K_I$ values of 46.3 $mmolmin^{-1}mg^{-1}$, 120 mM, and 24.4 mM, respectively. In a continuous packed-bed reactor, increasing the flow rate of the lactose solution decreased the conversion efficiency of lactose at different input lactose concentrations. Continuous operation of 11 days was conducted to investigate the stability of a long-term operation. The retained activity of the immobilized enzymes was 63% and the half-life of the immobilized enzyme was found to be 15 days.

• Journal of Korean Society of Environmental Engineers
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• v.32 no.12
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• pp.1126-1133
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• 2010

A two-stage biofilter was constructed and utilized to determine the removal efficiency when treating dynamic loading of a mixture of odorous compounds including benzene, toluene, p-xylene, ammonia and hydrogen sulfide. A yeast strain, Candida tropicalis, and a sulfur oxidizing bacterial (SOB) strain, Acidithiobacillus caldus sp., were immobilized in polyurethane media and packed in the two-stage biofilter. The experiment of dynamic loading variation was composed of (1) stepwise loading variation of all the odorous compounds (total EC test), (2) stepwise loading variation of each odorous compound, and (3) intermittent loading variation with 2-day-off and 3-day-on. The total EC test showed that the maximum elimination capacity was $61\;g/m^3/hr$ for total VOCs, and 5.2 and $9.1\;g/m^3/hr$ for ammonia and hydrogen, respectively. In addition, the inhibition between VOCs was observed when the loading of each individual VOC was varied. Especially the stepwise increase in toluene loading resulted in decreases of benzene and p-xylene removal efficiencies about 30% and 25%, respectively. However, the inhibition between organic and inorganic compounds was not observed. The intermittent loading variation with 2-day-off and 3-day-on showed that greater than 95% of the overall removal efficiency was restored in two days after the loading resumed. Consequently, the two-stage biofilter packed with immobilized microorganisms showed advantages over conventional biofilters for the simultaneous treatment of the mixture of organic and inorganic odorous compounds.