• 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.

• KSBB Journal
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• v.11 no.1
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• pp.77-85
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• 1996

The optimal initial pH for the ethanol production by Saccharomyces K35 was found to be 5.0, and about 80% of yield was obtained when 200g/$\ell$ of glucose was used as a substrate, which showed sugar tolerant. As the additives and cross-linking agent, the addition of 1.67%(w/v) Celite R-634 together with 0.33%(v/v) of glutaraldehyde(ACG bead) resulted in better stability, ethanol productivity and cell viability than Ca-alginate bead. Also, ACG bead seemed to be more resistant to phosphate ion than Ca-alginate bead, considering outgrowing cell concentration in the media. Scanning electron microscopic observation depicted that the surface of ACG bead was almost similar to the original state but not for Ca-alginate bead. When repealpd-batch culture was performed with Ca-alginate bead for 60 days in a 500m1 Erlenmeyer flask, ethanol and cell concentration were maintained about 138g/$\ell$-gel and 29~30g/$\ell$-gel, respectively, up to 40 days(7th run number), and then both were rapidly decreased. In the case of ACG bead, ethanol and cell concentration were maintained about 130~150g/$\ell$-gel and 32~35g/$\ell$-gel, respectively, up to 60days(10th run number). Cell viability was maintained about 70%, and outgrowing cell concentration was below 5.8% of total cell concentration.

• Journal of Korean Society of Environmental Engineers
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• v.32 no.2
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• pp.185-192
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• 2010

The objective of this study was to investigate the effectiveness of stabilization for army firing range soil highly contaminated with Pb (total Pb: 29,000 mg/kg) using calcined waste oyster shells. The calcination was conducted to activate quicklime from calcite. In order to evaluate the effectiveness of calcination, both natural oyster shells (NOS) and calcined oyster shells (COS) were applied to the Pb contaminated soil. Stabilization was conducted by mixing the contaminated soil with oyster shell media at 5-20 wt% and cured for 28 days. Following 28 days of curing, Pb leachability was measured based on the Korean Standard Test method (0.1 N HCl extraction). The treatment results showed that the COS treatment outperformed the NOS treatment. All of the NOS treatments failed to meet the Korean warning standard of 100 mg/kg. However, the Pb concentrations were significantly reduced to 47 mg/kg and 3 mg/kg upon 15 wt% and 20 wt% COS treatments, respectively which passed the Korean warning standard. Moreover, -#20 mesh materials were more effective than the -#10 mesh materials in effectively reducing Pb leachability. The scanning electron microscopy (SEM)-energy dispersive X-ray spectroscopy (EDX) results indicated that Pb immobilization was strongly linked to Al and Si.

• Korean Journal of Environmental Biology
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• v.41 no.1
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• pp.41-53
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• 2023

E171, a mixture of titanium dioxide, has been widely used as a food additive due to its whitening effect and low toxicity. However, it has been proven that E171 is no longer safe for public health. So far, there are insufficient studies on the toxic effects of E171 on organisms especially using standardized test methods. In this study, toxicity assessments of E171 to two aquatic species, water flea (Daphnia magna) and zebrafish (Danio rerio), were performed using modified standardized test methods based on the physicochemical properties of E171. The hydrodynamic diameter, polydispersity index, and turbiscan stability index (TSI) were measured to ensure the dispersion stability of E171 in exposure media during the test period. The EC₅₀ for immobilization of water flea was 141.7 mg L^-1 while zebrafish was not affected until 100 mg L^-1 of E171. Measurements of reactive oxygen species (ROS) and antioxidant enzyme activities confirmed that E171 induced oxidative stress, leading to the activation of superoxide dismutase and catalase in both water flea and zebrafish, although the expression of antioxidant enzyme genes differed between species. These results suggested the potential risk of E171 to aquatic organisms and provided toxicological insights into the impacts of E171 on the environment.

• 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.

• Korean Journal of Food Science and Technology
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• v.34 no.2
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• pp.255-262
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• 2002

This experiment was performed to improve the stability of Lactobacillus fermentum YL-3 as a poultry probiotics. The culture conditions that improve acid tolerance of L. fermentum YL-3 were investigated by changing several factors such as medium composition, temperature, anaerobic incubation and culture time. Also, L. fermentum YL-3 was encapsulated with alginate, calcium chloride and chitosan. The stable culture conditions of L. fermentum YL-3 were obtained in anaerobic incubation using MRS media without tween 80 for 20 hour at $42^{\circ}C$. The capsule after treatment with 1% chitosan was formed close membrane by a bridge bond. Immobilization of L. fermentum YL-3 in capsule was observed by confocal laser scanning microscopy, and cell viability was $2.0{\times}10^9\;CFU/g$ above the average. L. fermentum YL-3 capsule after acid treated at pH 2.0 for 3 hour survived about 40%, but those encapsulated with 1% chitosan survived about 65%. Survival rate of capsule stored at room temperature decreased about $2{\sim}3$ log cycle during 3 weeks, but viability of capsule stored at $4^{\circ}C$ during 3 weeks maintained almost $10^8\;CFU/g$ levels.