• Korean Journal of Microbiology
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• v.46 no.1
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• pp.46-51
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• 2010

Coenzyme Q10 (CoQ10) is an essential lipid-soluble component of membrane-bound electron transport chains. CoQ10 is involved in several aspects of cellular metabolism and is increasingly being used in therapeutic applications for several diseases. Despite the recent accomplishments in metabolic engineering of Escherichia coli for CoQ10 production, the production levels are not yet competitive with those by fermentation or isolation. So we tested several microorganisms obtained from the KCTC of Biological Resource Center to find novel sources of strain-development for CoQ10-production. Then we selected two strains, Paracoccus denitrificans (KCTC 2530) and Asaia siamensis (KCTC 12914), and tested to optimize the CoQ10 production conditions. Among the carbon sources tested, CoQ10 production was the highest when fructose was supplied about 4% concentration. Yeast extract produced the highest CoQ10 production about 2% concentration. The highest CoQ10 production was obtained at pH 6.0 for P. denitrificans and pH 8.0 for A. siamensis. And two strains showed the highest CoQ10 production at $30^{\circ}C$, but the highest DCW was obtained at $37^{\circ}C$. In the fed-batch culture, P. denitrificans yielded $14.34{\pm}0.473$ mg and A. siamensis yielded $12.53{\pm}0.231$ mg of final CoQ10 production.

• Journal of Ecology and Environment
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• v.32 no.4
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• pp.257-265
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• 2009

We estimated monthly and annual secondary productivity of pelagic zooplankton in Lake Paldang and Lake Cheongpyong. Secondary productivity was calculated by combining estimated zooplankton biomass and biomass-specific productivity for each site and depth from March to November 2008. In addition to somatic production, we measured production of eggs and exuviae for three dominant species: Daphnia galeata, Bosmina longirostris, Cyclops sp. In terms of biomass, B. longirostris was dominant in Lake Paldang in April and May, B. longirostris showed explosive biomass growth, especially in May. In June and July, B. longirostris and D. galeata were both dominant. Lake Cheongpyeong showed much lower zooplankton biomass than Lake Paldang. In August, there was little or no biomass in both lakes probably due to heavy rain. The Gyeongan River contributed most of the secondary productivity and B. longirostris contributed the most secondary productivity in Lake Paldang. D. galeata also contributed in the Gyeongan River, the South Han River and at the Paldang Dam in spring and fall. Overall, Lake Cheongpyeong showed lower secondary productivity than Lake Paldang. B. longirostris made the largest contribution to secondary productivity in the Cheongpyeong Dam area while D. galeata contributed the most near Nami Island. Somatic production constituted ~80% of the total secondary productivity (the sum of somatic, egg and exuvia production) for D. galeata and B. longirostris. Although production-to-biomass (P/B) ratios were usually <<1 B. longirostris sometimes showed very high P/B ratios, probably due to fish predation. D. galeata showed much lower P/B ratios than B. longirostris after the summer at most sites.

• Journal of Microbiology and Biotechnology
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• v.26 no.2
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• pp.241-247
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• 2016

Natamycin is a widely used antifungal antibiotic. For natamycin biosynthesis, the gene pimE encodes cholesterol oxidase, which acts as a signalling protein. To confirm the positive effect of the gene pimE on natamycin biosynthesis, an additional copy of the gene pimE was inserted into the genome of Streptomyces gilvosporeus 712 under the control of the ermE* promoter (p_ermE*) using intergeneric conjugation. Overexpression of the target protein engendered 72% and 81% increases in the natamycin production and cell productivity, respectively, compared with the control strain. Further improvement in the antibiotic production was achieved in a 1 L fermenter to 7.0 g/l, which was a 153% improvement after 120 h cultivation. Exconjugants highly expressing pimE and pimM were constructed to investigate the effects of both genes on the increase of natamycin production. However, the co-effect of pimE and pimM did not enhance the antibiotic production obviously, compared with the exconjugants highly expressing pimE only. These results suggest not only a new application of cholesterol oxidase but also a useful strategy to genetically engineer natamycin production.

• Journal of Microbiology and Biotechnology
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• v.34 no.3
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• pp.700-709
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• 2024

Polyhydroxybutyrate (PHB) production from lignocellulosic biomass is economically beneficial. Because lignocellulosic biomass is a mixture rich in glucose and xylose, Escherichia coli, which prefers glucose, needs to overcome glucose repression for efficient biosugar use. To avoid glucose repression, here, we overexpressed a xylose regulator (xylR) in an E. coli strain expressing bktB, phaB, and phaC from Cupriavidus necator and evaluated the effect of xylR on PHB production. XylR overexpression increased xylose consumption from 0% to 46.53% and produced 4.45-fold more PHB than the control strain without xylR in a 1% sugar mixture of glucose and xylose (1:1). When the xylR-overexpressed strain was applied to sugars from lignocellulosic biomass, cell growth and PHB production of the strain showed a 4.7-fold increase from the control strain, yielding 2.58 ± 0.02 g/l PHB and 4.43 ± 0.28 g/l dry cell weight in a 1% hydrolysate mixture. XylR overexpression increased the expression of xylose operon genes by up to 1.7-fold. Moreover, the effect of xylR was substantially different in various E. coli strains. Overall, the results showed the effect of xylR overexpression on PHB production in a non-native PHB producer and the possible application of xylR for xylose utilization in E. coli.

• Journal of Plant Biotechnology
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• v.37 no.3
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• pp.262-268
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• 2010

Antibodies are powerful and versatile tools to play a critical role in the diagnosis and treatment of many diseases. Their application has been enhanced significantly with the advanced recombinant DNA and heterologonous expression technologies, allowing to produce immunotherapeutic proteins with improved biofunctional properties. However, with currently available technologies, mammalian cell-based therapeutic antibody production, as an alternative for production in humans and animals, is often not plentiful for passive immunotherapeutics in treatment of many diseases. Recently, plant expression systems for therapeutic antibodies have become well-established. Thus, plants have been considered to provide an attractive alternative production system for therapeutic antibodies, as plants have several advantages such as the lack of human pathogens, and low cost of upstream production and flexible scale-up of highly valuable recombinant glycoproteins. Recent advances in modification of posttranslational processing for human-like glycosylation in transgenic plants will make it possible that plant can become a suitable protein expression system over the animal cellbased current production system. This review will discuss recent advances in plant expression technology and issues for their application to therapeutic antibody production.

• KSBB Journal
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• v.28 no.4
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• pp.260-268
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• 2013

Transgenic plant cell cultures are an attractive expression system for the production of industrial and pharmaceutical proteins because of their advantages in safety and low production cost. Human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) was produced and secreted when sugar was depleted in culture medium by transgenic rice cell lines (Oryza sativa L.) using RAmy3D promoter. Due to the production of the target protein by sugar depletion, concomitant occurrence of cell death is inevitable. For that reason, inhibition of cell death for enhancing productivity was necessary for the production period without energy sources. Supplementation of 0.1 mM sodium nitroprusside improved cell viability by 1.4-fold and maximum hCTLA4Ig production by 1.3-fold compared to those of control. Addition of 1 and 10 mM glutathione, N-acetylcysteine (NAC), and nicotinamide inhibited apoptotic-like programmed cell death by decreasing the activity of reactive oxygen species. Production hCTLA4Ig was enhanced 1.4-, 1.25-, and 1.15-fold with 10 mM NAC, 1 mM NAC, and 1 mM glutathione, respectively. In addition, it was found that the supplementation of NAC enhanced the cell viability.

• Journal of Ecology and Environment
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• v.44 no.4
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• pp.229-240
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• 2020

Background: Genus Quercus is a successful group that has occupied the largest area of forest around the world including South Korea. The acorns are an important food source for both wild animals and humans. Although the reproductive characteristics of this genus are highly variable, it had been rarely studied in South Korea. Therefore, in Seoraksan and Odaesan National Parks (i) we measured the acorn production of Quercus mongolica, an overwhelmingly dominant species in South Korea, for 3 years (2017-2019), (ii) evaluated the spatial-temporal variation of acorn production, and (iii) analyzed the effects of oak- and site-related variables on the acorn production. Results: The annual acorn production of Q. mongolica increased 36 times from 1.2 g m-2 in 2017 to 43.2 g m-2 in 2018, and decreased to 16.7 g m-2 in 2019, resulting in an annual coefficient of variation of 104%. The coefficient of spatial variation was high and reached a maximum of 142%, and the tree size was the greatest influencing factor. That is, with an increase in tree size, acorn production increased significantly (2018 F = 16.3, p < 0.001; 2019 F = 8.2, p < 0.01). Elevation and slope also significantly affected the production in 2019. However, since elevation and tree size showed a positive correlation (r = 0.517, p < 0.001), the increase in acorn production with increasing elevation was possibly due to the effect of tree size. The acorn production of Odaesan for 3 years was 2.2 times greater than that of Seoraksan. This was presumed that there are more distribution of thick oak trees and more favorable site conditions such as deep soil A-layer depth, high organic matter, and slower slopes. Conclusion: As reported for other species of the genus Quercus, the acorn production of Q. mongolica showed large spatial and annual variations. The temporal variability was presumed to be a weather-influenced masting, while the spatial variability was mainly caused by oak tree size.

• Journal of Microbiology and Biotechnology
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• v.25 no.10
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• pp.1653-1659
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• 2015

Epothilone, which is produced by the myxobacterium Sorangium cellulosum, contributes significant value in medicinal development. However, under submerged culture conditions, S. cellulosum will accumulate to form bacterial clumps, which hinder nutrient and metabolite transportation. Therefore, the production of epothilone by liquid fermentation is limited. In this study, diatomite-based porous ceramics were made from diatomite, paraffin, and poremaking agent (saw dust). Appropriate methods to modify the porous ceramics were also identified. After optimizing the preparation and modification conditions, we determined the optimal prescription to prepare high-performance porous ceramics. The structure of porous ceramics can provide a solid surface area where S. cellulosum can grow and metabolize to prevent the formation of bacterial clumps. S. cellulosum cells that do not form clumps will change their erratic metabolic behavior under submerged culture conditions. As a result, the unstable production of epothilone by this strain can be changed in the fermentation process, and the purpose of increasing epothilone production can be achieved. After 8 days of fermentation under optimized conditions, the epothilone yield reached 90.2 mg/l, which was increased four times compared with the fermentation without porous ceramics.

• Journal of Microbiology and Biotechnology
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• v.19 no.2
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• pp.136-139
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• 2009

Avermectin and its analogs are major commercial antiparasitic agents in the fields of animal health, agriculture, and human infections. Previously, comparative transcriptome analysis between the low-producer S. avermitilis ATCC31267 and the high-producer S. avermitilis ATCC31780 using a S. avermitilis whole genome chip revealed that 50 genes were overexpressed at least two-fold higher in S. avermitilis ATCC31780. To verify the biological significance of some of the transcriptomics-guided targets, five putative regulatory genes were individually cloned under the strong-and-constitutive promoter of the Streptomyces expression vector pSE34, followed by the transformation into the low-producer S. avermitilis ATCC31267. Among the putative genes tested, three regulatory genes including SAV213, SAV3818, and SAV4023 exhibited stimulatory effects on avermectin production in S. avermitilis ATCC31267. Moreover, overexpression of SAV3818 also stimulated actinorhodin production in both S. coelicolor M145 and S. lividans TK21, implying that the SAV3818, a putative TetR-family transcriptional regulator, could be a global upregulator acting in antibiotic production in Streptomyces species.

• Journal of Microbiology and Biotechnology
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• v.18 no.12
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• pp.1927-1931
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• 2008

In this study, the enzymatic process for biodiesel production was optimized using a mixture of immobilized Rhizopus oryzae and Candida rugosa lipases. The optimal temperature and agitation speed for biodiesel production were $45^{\circ}C$ and 300 rpm, respectively. The optimal ratio of R. oryzae and C. rugosa lipases in the mixture was 3:1 (w:w). When 3 mmol of methanol was the initial reaction medium and 3 mmol of methanol was added every 1.5 h during biodiesel production, biodiesel conversion was over 98% at 4 h. In addition, when the immobilized lipase mixture was reused, biodiesel conversion exceeded 80% after 5 reuses.