• Molecules and Cells
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• v.24 no.2
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• pp.276-282
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• 2007

Protein phosphorylation is one of the major mechanisms by which eukaryotic cells transduce extracellular signals into intracellular responses. Calcium/calmodulin ($Ca^{2+}/CaM$)-dependent protein phosphorylation has been implicated in various cellular processes, yet little is known about $Ca^{2+}/CaM$-dependent protein kinases (CaMKs) in plants. From an Arabidopsis expression library screen using a horseradish peroxidase-conjugated soybean calmodulin isoform (SCaM-1) as a probe, we isolated a full-length cDNA clone that encodes AtCK (Arabidopsis thaliana calcium/calmodulin-dependent protein kinase). The predicted structure of AtCK contains a serine/threonine protein kinase catalytic domain followed by a putative calmodulin-binding domain and a putative $Ca^{2+}$-binding domain. Recombinant AtCK was expressed in E. coli and bound to calmodulin in a $Ca^{2+}$-dependent manner. The ability of CaM to bind to AtCK was confirmed by gel mobility shift and competition assays. AtCK exhibited its highest levels of autophosphorylation in the presence of 3 mM $Mn^{2+}$. The phosphorylation of myelin basic protein (MBP) by AtCK was enhanced when AtCK was under the control of calcium-bound CaM, as previously observed for other $Ca^{2+}/CaM$-dependent protein kinases. In contrast to maize and tobacco CCaMKs (calcium and $Ca^{2+}/CaM$-dependent protein kinase), increasing the concentration of calmodulin to more than $3{\mu}M$ suppressed the phosphorylation activity of AtCK. Taken together our results indicate that AtCK is a novel Arabidopsis $Ca^{2+}/CaM$-dependent protein kinase which is presumably involved in CaM-mediated signaling.

• Journal of Microbiology and Biotechnology
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• v.30 no.11
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• pp.1720-1728
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• 2020

We have previously characterized AprESJ4, the major fibrinolytic enzyme from Bacillus subtilis SJ4 (Yao et al., 2019). During that study, we observed a 68 kDa protein with fibrinolytic activity. In this study, we cloned the gene (vprSJ4) encoding the 68 kDa protein, a mature Vpr and minor protease secreted by Bacillus species. vprSJ4 encodes a preproenzyme consisting of 810 amino acids (aa) including signal sequence (28 aa) and prosequence (132 aa). The mature enzyme (650 aa) has a predicted molecular weight of 68,467.35. Unlike Vprs from other B. subtilis strains, VprSJ4 has 4 additional amino acids (DEFA) at the C-terminus. vprSJ4 was overexpressed in Escherichia coli. PreproVprSJ4 was localized in inclusion bodies, and subjected to in vitro renaturation and purification by an affinity column. SDS-PAGE and western blot showed that autoprocessing of preproVprSJ4 occurred and 68 kDa and smaller proteins were produced. The optimum pH and temperature of the recombinant VprSJ4 were pH 7.0 and 40℃, respectively. Kinetic parameters of recombinant VprSJ4 were measured by using an artificial substrate, N-succinyl-ala-ala-pro-phe-p-nitroanilide. Coexpression of vprSJ4 and aprESJ4 using pHY300PLK increased the fibrinolytic activity a further 117% when compared with aprESJ4 single expression using the same vector in B. subtilis WB600.

• Journal of Microbiology and Biotechnology
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• v.16 no.1
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• pp.149-152
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• 2006

Leuconostoc mesenteroides SY1, a strain isolated from cabbage Kimchi, was transformed with pCW4, a shuttle vector based on a cryptic plasmid from Lactobacillus paraplantarum C7. $\alpha-Amylase$ gene, amyL, from Bacillus licheniformis was cloned into pCW4, resulting in $pCW4T{\alpha},\;and\;pCW4T{\alpha}$ was introduced into SY1 by electroporation. Transformation efficiency was $10^2cells/{\mu}g$ plasmid DNA. L. mesenteroides cells harboring $pCW4T{\alpha}$ did not show amylase activity, although amyL transcript was synthesized as determined by slot blot experiment. $pCW4T{\alpha}$ was stably maintained in SY1 in the presence of erythromycin (Em, $5\;{\mu}g/ml$) but rapidly lost when Em was omitted. Less than $1\%$ of the cells maintained $pCW4T{\alpha}$ after 5 days at $30^{\circ}C$.

• Molecules and Cells
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• v.43 no.1
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• pp.76-85
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• 2020

MARCH5 is a RING finger E3 ligase involved in mitochondrial integrity, cellular protein homeostasis, and the regulation of mitochondrial fusion and fission. To determine the function of MARCH5 during development, we assessed transcript expression in zebrafish embryos. We found that march5 transcripts were of maternal origin and evenly distributed at the 1-cell stage, except for the mid-blastula transition, with expression predominantly in the developing central nervous system at later stages of embryogenesis. Overexpression of march5 impaired convergent extension movement during gastrulation, resulting in reduced patterning along the dorsoventral axis and alterations in the ventral cell types. Overexpression and knockdown of march5 disrupted the organization of the developing telencephalon and diencephalon. Lastly, we found that the transcription of march5 was tightly regulated by the transcriptional regulators CHOP, C/EBPα, Staf, Znf143a, and Znf76. These results demonstrate the essential role of March5 in the development of zebrafish embryos.

• Food Science and Biotechnology
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• v.18 no.5
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• pp.1273-1278
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• 2009

This study aimed to evaluate the potential allergenicity of Cry proteins in insect-resistant genetically modified (GM) maizes (Bt11, MON810, and MON863) using serum screening tests. Serum samples were obtained from Korean children (0-15 years old) with allergic symptoms who had positive maize-specific IgE. The levels of serum specific IgE was measured by the Phadia ImmunoCAP system and considered as positive when they are 0.35 kU/L or higher. Cry proteins (Cry1Ab in Bt11, mCry1Ab in MON810, and Cry3Bb1 in MON863) were expressed in Escherichia coli and purified for serum screening. The reactivity of purified Cry proteins was confirmed by IgE immunoblots in 50 patients (maize-sensitized patients). There was no reaction between Cry proteins and sera from maize-sensitized patients. Our results suggest that these Cry proteins are not likely to cause allergic reactions. Further studies using more sera from patients with true clinical allergies are needed to evaluate the potential allergenicity of novel proteins in GM maize.

• East Asian Journal of Business Economics (EAJBE)
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• v.10 no.4
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• pp.75-90
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• 2022

Purpose - This study aims to study the activation pattern of declining industries by applying the Gompertz growth model using available resources based on the theory of industrial life cycle, classifying declining industries among Korean manufacturing industries, and identifying resource input characteristics. Research design and methodology - This study was conducted by combining the Gompertz growth model that predicts the limit of output based on available resources under the industrial life cycle theory. Using Gompertz model, this study analyzed the life cycle of 39 Korean manufacturing industries from the perspective of domestic production, number of employees, and fixed assets Results - According to a life cycle analysis of 39 manufacturing industries in Korea, the computer, textile, and shoe industries were classified as declining industries. Among them, research on resource input characteristics on the shoe industry showed that domestic production and the number of employees decreased, while the proportion of domestic R&D personnel and the number of research departments gradually increased. Conclusion - Among the declining industries in Korea, the shoe industry is considered to revitalize the industry, that is, to extend the life of the declining industry by offshoring its production site and improving constitution with a "R&D center for global" support.

• Preventive Nutrition and Food Science
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• v.10 no.2
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• pp.118-121
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• 2005

Methanol extracts were prepared from loquat leaves (2 kg) and successively fractionated with hexane, chloroform, ethyl acetate, butanol, and water solvents. The ethyl acetate fraction exhibited the highest antioxidative and, antimicrobial activities. Therefore, the ethyl acetate fraction was purified and a chemical structure was identified by $^1H$ and $^{13}C-NMR$ spectra, FT-IR and EI/MS spectroscopies. The isolated antioxidative and antimicrobial substance was identified as epicatechin.

• Journal of Microbiology and Biotechnology
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• v.12 no.6
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• pp.921-928
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• 2002

Microorganism producing extracellular CFTase was isolated from soil and designated as Bacillus polymyxa MGL21. The gene encoding the CFTase (cft) from B. polymyxa MGL21 was cloned and sequenced. The ORF of the cf gene was composed of 3,999 nucleotides, encoding a protein (1,333 amino acids) with a predicted molecular mass of 149,375 Da. Sequence analysis indicated that CFTase was divided into five distinct regions. CFTase contained three regions of repeat sequences at the N-terminus and C-terminus. The endo-inulinase region of homology (ERH) of CFTase was similar to that of Pseudomonas mucidolens endo-inulinase ($50\%$ identity, 259 amino acids). Furthermore, CFTase possessed a highly conserved core region, which is considered to be functional for the hydrolysis reaction of inulin. The cft gene was expressed in a His-tagged form in Escherichia coli cells, and the His-tagged CFTase was purified to homogeneity. The optimal temperature and pH for CFTase activity were found to be $50^{\circ}C$ and 9.0, respectively. The enzyme activity was completely inhibited by 10 mM $Ag^+\;and\;Cu^2+$. Thin-layer chromatography analyses indicated that CFTase catalyzed not only the cyclization reaction ut also disproportionation and hydrolysis reactions as well.

• Korean journal of food and cookery science
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• v.28 no.2
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• pp.183-193
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• 2012

In this study, data was collected on the dietary life education at elementary schools according to the Food Education Support Act using elementary school teachers as subjects. A survey was conducted on 258 elementary school teachers at seven elementary schools located in Seoul, Gyeonggi, and Daegu, Korea from April 14-17, 2010. Most teachers reported that dietary life education was very much needed. The appropriate individuals to teach dietary life education were nutrition teachers, charge teachers, and dietary life education tutors, in that order. The most appropriate time to conduct dietary life education was during dietary life-related subject hours, dietary life education hours, food service hours, and discretionary and extracurricular activities, in that order. The most effective materials and methods used for dietary life education were projection materials and dietary life-related special lectures. Efficient dietary life education methods can be used to develop dietary life education programs. Based on these results, elementary school teachers recognize that dietary life education is very much needed and believe that it would be desirable to conduct dietary life education using nutrition teachers in cooperation with other related teachers.

• Journal of Microbiology and Biotechnology
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• v.20 no.8
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• pp.1226-1229
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• 2010

Glyceollins, one of the inducible phytoalexins produced by plants, were induced in a number of varieties of Korean soybean through fungal infection. Of the tested soybean varieties, Tae-Kwang, though not the most productive, was found to be currently the most suitable for the induction of glyceollins. Amongst the fungal species, Rhizopus microsporus var. oligosporus was seen to be the most effective elicitor. Halved soybean seeds produced glyceollins upon fungal infection; however, chopped soybeans and homogenized soybeans did not produce significant quantities of glyceollins.