• Journal of Microbiology and Biotechnology
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• v.31 no.1
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• pp.163-170
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• 2021

Enzyme replacement therapy for lysosomal storage diseases usually requires recombinant enzymes containing mannose-6-phosphate (M6P) glycans for cellular uptake and lysosomal targeting. For the first time, a strategy is established here for the in vitro mannosyl-phosphorylation of high-mannose type N-glycans that utilizes a recombinant Mnn14 protein derived from Saccharomyces cerevisiae. Among a series of N-terminal- or C-terminal-deleted recombinant Mnn14 proteins expressed in Pichia pastoris, rMnn1477-935 with deletion of N-terminal 76 amino acids spanning the transmembrane domain (46 amino acids) and part of the stem region (30 amino acids), showed the highest level of mannosyl-phosphorylation activity. The optimum reaction conditions for rMnn1477-935 were determined through enzyme assays with a high-mannose type N-glycan (Man8GlcNAc2) as a substrate. In addition, rMnn1477-935 was shown to mannosyl-phosphorylate high-mannose type N-glycans (Man7-9GlcNAc2) on recombinant human lysosomal alpha-glucosidase (rhGAA) with remarkably high efficiency. Moreover, the majority of the resulting mannosyl-phosphorylated glycans were bis-form which can be converted to bis-phosphorylated M6P glycans having a superior lysosomal targeting capability. An in vitro N-glycan mannosyl-phosphorylation reaction using rMnn1477-935 will provide a flexible and straightforward method to increase the M6P glycan content for the generation of "Biobetter" therapeutic enzymes.

• Pediatric Infection and Vaccine
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• v.7 no.1
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• pp.83-93
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• 2000

Purpose : Urabe AM-9 strain was known to be associated with increased aseptic meningitis. The reason for high incidence of vaccine-associated meningitis was known that nucleotide(nt) substituted form G to A at position 1081 of the hemagglutinin-neuraminidase(HN) gene and therefore, glutamic acid changed to lysine at amino acid 335. We assessed by comparing nt sequence of the HN gene form Urabe AM-9 strain with wild strain and documented the correlation between nt substitution and vaccine-associated meningitis. Methods : Two lots of Urabe AM-9 vaccine distributed in Korea and mumps wild strains isolated from 1998 through 1999 were analysed. Analysis was made by nt sequencing following amplification of HN gene by RT-PCR. Results : Nucleotide substitution at position 343, 1476, 1570 was not found in both Urabe AM-9 vaccines and wild strains. But analysis of vaccine strains and wild strains isolated from patients revealed substitution from G to A at nt 1081 of the HN gene. Therefore, it encodes lysine instead of glutamic acid at amino acid 335. There was no mixture from of G and A at nt 1081. Nt at 1470 of one lot of Urabe AM-9 vaccines changed from C to A after Vero cell passage. Nt at 1727 of vaccines and wild strains was substituted A to G, so it encodes glycine instead of aspartic acid. Conclusion : Nucleotide analysis of HN gene revealed that nt 1081 of Urabe AM-9 vaccines and wild strains had wild type AAA($Lys^{335}$) instead of variant type GAA($Glu^{335}$). The results of this study suggest that there was a probability of vaccine-associated meningitis due to Urabe AM-9 in Korea before. But incidence of actual side effect was not evaluated because there was no reporting system in Korea.

• Journal of Environmental Science International
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• v.23 no.5
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• pp.839-852
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• 2014

The Ministry of Environment (MOE) has made more effort in managing point source pollution rather than in nonpoint source pollution in order to improve water quality of the four major rivers. However, it would be difficult to meet water quality targets solely by managing the point source pollution. As a result of the comprehensive measures established in 2004 under the leadership of the Prime Minister's Office, a variety of policies such as the designation of control areas to manage nonpoint source pollution are now in place. Various action plans to manage nonpoint source pollution have been implemented in the Soyang-dam watershed as one of the control areas designed in 2007. However, there are no tools to comprehensively assess the effectiveness of the action plans. Therefore, this study would assess the action plans (especially, BMPs) designed to manage Soyang-dam watershed with the WinHSPF and the CE-QUAL-W2. To this end, we simulated the rainfall-runoff and the water quality (SS) of the watershed and the reservoir after conducting model calibration and the model validation. As the results of the calibration for the WinHSPF, the determination coefficient ($R^2$) for the flow (Q, $m^3/s$) was 0.87 and the $R^2$ for the SS was 0.78. As the results of the validation, the former was 0.78 and the latter was 0.67. The results seem to be acceptable. Similarly, the calibration results of the CE-QUAL-W2 showed that the RMSE for the water level was 1.08 and the RMSE for the SS was 1.11. The validation results(RMSE) of the water level was 1.86 and the SS was 1.86. Based on the daily simulation results, the water quality target (turbidity 50 NTU) was not exceeded for 2009~2011, as results of maximum turbidity in '09, '10, and '11 were 3.1, 2.5, 5.6 NTU, respectively. The maximum turbidity in the years with the maximum, the minimum, and the average of yearly precipitation (1982~2011) were 15.5, 7.8, and 9.0, respectively, and therefore the water quality target was satisfied. It was discharged high turbidity at Inbuk, Gaa, Naerin, Gwidun, Woogak, Jeongja watershed resulting of the maximum turbidity by sub-basins in 3years(2009~2011). The results indicated that the water quality target for the nonpoint source pollution management should be changed and management area should be adjusted and reduced.

• Journal of Wetlands Research
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• v.21 no.spc
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• pp.98-106
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• 2019

The coefficient and exponent of the MUSLE(Modified Universal Soil Loss Equation) runoff factor in the SWAT(Soil and Water Assessment Tool) model are 11.8 and 0.56 respectively, which are equally applied to the estimation of soil erosion regardless of land use. they could derive overestimation or underestimation of soil erosion, which can cause problems in the selection of soil erosion-vulnerable area and evaluation of reduction management. However, there are no studies about the estimation of coefficients and exponent for the MUSLE runoff factor by land use and their applicability to the SWAT model. Thus, in order to predict soil erosion and sediment behavior accurately through SWAT model, it is necessary to estimate the coefficient and exponent of the MUSLE runoff factor by land use and evaluate its applicability. In this study, the coefficient and exponent of MUSLE runoff factor by land use were estimated for Gaa-cheon Watershed, and the differences in soil erosion and sediment from SWAT model were analyzed. The coefficient and exponent of runoff factor estimated by this study well reflected the characteristics of soil erosion in domestic highland watershed. Therefore, in order to apply the MUSLE which developed based on observed data of US agricultural basin to the domestic watershed, it is considered that a sufficient modification and supplementation process for the coefficient and exponent of the MUSLE runoff factor depending on land use is necessary. The results of this study can be used as a basic data for selecting soil erosion vulnerable area in the non-point source management areas and establishing and evaluating soil erosion reduction management.