• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.3
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• pp.225-229
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• 2005

To investigate impediments to plasmid transformation in Brevibacterium flavum BF4 and B. lactofermentum BL1, cell surface barriers were determined by measuring growth inhibition whilst enzymatic barriers were determined by comparing DNA methylation properties. B. lactofermentum was more sensitive to growth inhibition by glycine than B. flavum. Release of cellular proteins during sonication was more rapid for B. lactofermentum than for B. flavum. Plasmid DNA (pCSL 17) isolated from B. flavum transformed recipient $McrBC^+$ strains of Escherichia coli with lower efficiency than $McrBC^-$. McrBC digestion of this DNA confirmed that B. flavum contain methylated cytidines in the target sequence of McrBc sequences but B. lactofermentum contained a different methylation pattern. DNA derived from the B. lactofermentum transformed recipient $EcoKR^+$ strains of E. coli with lower efficiency than $EcoKR^-$, indicating the presence of methylated adenosines in the target sequence of EcoK sequences. The present data describe the differences in the physical and enzymatic barriers between two species of corynebacteria and also provide some insight into the successful foreign gene expression in corynebacteria.

• BMB Reports
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• v.30 no.6
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• pp.438-442
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• 1997

${\gamma}-Glutamylamine$ cyclotransferase was purified to homogeneity from soybean (Glycine max) seeds. To our knowledge, it is the first purification of the enzyme from plant origins. The molecular weight of the enzyme estimated by Sephacryl S-300 gel filtration and SDS-PAGE was 27,000, indicating that the enzyme is a monomer. The optimal pH for activity was 8.6. The Km value for ${\gamma}-glutamyldansylcadaverine$ was 11 ${\mu}M$. The enzymatic activity was substantially inhibited by the addition of p-chloromercuribenzoate and partially inhibited by the $Cu^{2+}$ ion. However, neither other modification reagents nor other divalent metal ions affected the enzymatic activity. The comparison between the enzymatic activities of seed extracts treated with cycloheximide and control extracts, and the detection of the same single protein band by western blot analysis at the dormant stage without inhibition with distilled water indicate that ${\gamma}-Glutamylamine$ cyclotransferase is already present at the dormant stage and gradually activated during germination in soybean seeds.

• Korean Chemical Engineering Research
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• v.62 no.2
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• pp.147-152
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• 2024

Most medical sensors are disposable products. In order to reduce inspection and diagnosis costs, it is more important to develop the inexpensive electrode materials. We fabricated the CuO NPs/PANI/E-PGE as an electrode material for disposable electrochemical sensors and applied it to a non-enzymatic glucose sensor. For surface activation of PGE, pretreatment was performed using chemical and electrochemical methods, respectively. Electrochemical properties according to the pretreatment method were analyzed through chronoamperometry (CA), cyclic voltammetry (CV) and electrochemical impedance (EIS). From these analytical results, the electrochemically pretreated PGE (E-PGE) was finally adopted. The non-enzymatic glucose sensor based on CuO NPs/PANI/E-PGE shows sensitivity of 239.18 mA/mM×cm² (in a linear range of 0.282~2.112 mM) and 36.99 mA/mM×cm² (3.75423~50 mM), detection limit of 17.6 μM and good selectivity. Based on the results of this study, it was confirmed that the modified PGE is a high-performance electrode material. Therefore, these electrodes can be applied to a variety of disposable sensors.

• Journal of Microbiology and Biotechnology
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• v.26 no.5
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• pp.837-845
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• 2016

A novel endodextranase isolated from Paenibacillus sp. was found to produce isomaltotetraose and small amounts of cycloisomaltooligosaccharides with a degree of polymerization of 7-14 from dextran. To determine the active site, the enzyme was modified with 1-ethyl-3-[3-(dimethylamino)-propyl]-carbodiimide (EDC) and α-epoxyalkyl α-glucosides (EAGs), an affinity labeling reagent. The inactivation followed pseudo first-order kinetics. Kinetic analysis and chemical modification using EDC and EAGs indicated that carboxyl groups are essential for the enzymatic activity. Three Asp and one Glu residues were identified as candidate catalytic amino acids, since these residues are completely conserved across the GH family of 66 enzymes. Replacement of Asp189, Asp340, or Glu412 completely abolished the enzyme activity, indicating that these residues are essential for catalytic activity.

• BMB Reports
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• v.48 no.8
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• pp.431-437
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• 2015

Notch signaling plays a pivotal role in cell fate determination, cellular development, cellular self-renewal, tumor progression, and has been linked to developmental disorders and carcinogenesis. Notch1 is activated through interactions with the ligands of neighboring cells, and acts as a transcriptional activator in the nucleus. The Notch1 intracellular domain (Notch1-IC) regulates the expression of target genes related to tumor development and progression. The Notch1 protein undergoes modification after translation by posttranslational modification enzymes. Phosphorylation modification is critical for enzymatic activation, complex formation, degradation, and subcellular localization. According to the nuclear cycle, Notch1-IC is degraded by E3 ligase, FBW7 in the nucleus via phosphorylation-dependent degradation. Here, we summarize the Notch signaling pathway, and resolve to understand the role of phosphorylation in the regulation of Notch signaling as well as to understand its relation to cancer. [BMB Reports 2015; 48(8): 431-437]

• Journal of Microbiology and Biotechnology
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• v.11 no.1
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• pp.118-123
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• 2001

The alginate lyase A1-III gene of Sphingomonas species A1 is composed of 1,077 nucleotides, encoding a protein (359 amino acids) with a molecular mass of 40,322 Da. Recombinant A1-III expressed in Escherichia coli exhibited the same full enzymatic activity as native A1-III. In order to identify the critical residue for activity, a site-directed mutation was introduced into the A1-III gene (H192A, His192->Ala). Recombinant A1-III (H192A) exhibited a significant decrease in enzyme activity (one-thirty thousandth of that of A1-III), without any conformational change, as detected by the CD spectra in the far UV region. Also, the chemical modification of wild-type A1-III with methyl 4-nitro benzene sulfonate resulted in a 40% decrease from the initial activity, whereas the same modification of A1-III (H192A) produced no change in the activity. The role of His192 on the catalytic process was also explored based on a model of A1-III docked with mannuronic acid into the active site.

• Journal of Powder Materials
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• v.14 no.4
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• pp.256-260
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• 2007

The surface of magnetite ($Fe_{3}O_{4}$) nanoparticles prepared by coprecipitation method was modified by carboxylic acid group of poly(3-thiophenacetic acid (3TA)) and meso-2,3-dimercaptosuccinic acid (DMSA). Then the lysozyme protein was immobilized on the carboxylic acid group of the modification of the magnetite nanoparticles. The magnetite nanoparticles are spherical and the particle size is approximately 10 nm. We measured quantitative dispersion state by dispersion stability analyzer for each $Fe_{3}O_{4}$ nanoparticles with and without surface modification. The concentration of lysozyme on the modified magnetite nanoparticles was also investigated by a UV-Vis spectrometer and compared to that of magnetite nanoparticles without surface modification. The functionalized magnetite particles had higher enzymatic capacity and dispersion stability than non-functionalized magnetite nanoparticles.

• Korean Journal of Food Science and Technology
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• v.19 no.3
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• pp.212-219
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• 1987

The rec-assay on Bacillus subtilis strains H17$({Rec}^+)$ and M45$({Rec}^-)$, the Ames test with modification of preincubation on Salmonella typhimurium TA98 and TA100 and DNA-breaking test on double strand calfthymus DNA were carried out using enzymatically browned substances obtained from the reaction of Prunus salicina (Red) enzyme and polyphenols. The spore rec-assay of enzymatic browning reaction products of pyrogallol, hydroxyhydroquinone. 3,4-dihydrohyoluene and chlorogenic acid showed non-mutagenic activity The spore rec-assay showed a little influence of ${Zn}^{2+}$ and ${Ni}^{2+}$ on the action of four kinds of enzymatic browning reaction products. The enzymatic browning reaction products of polyphenols did not show DNAbreaking activity. ${Cu}^{2+}$ of various metal ions influenced on DNA-breaking of enzymatic browning reaction products of pyrogallol. However, enzymatic browning reaction products of chlorogenic acid inhibited on DNA-breaking activity. Four kinds of enzymatic browning reaction products showed non-mutagenic activity on Salmonella typhimurium TA98 and TA100 with S-9 mix. In the mutagenicity on Salmonella typhimurium TA98 and TA100 with S-9 mix in the presence of benzo$({\alpha})$pyrene which is the carcinogenic substances, four kinds of enzymatic browning reaction products showed desmutagenic activity.

• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.2
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• pp.262-267
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• 2013

The purpose of this research is to minimize the loss of nutrients in carrots (Daucus carota var. sativa). A protopectinase was used to enzymatically macerated and separate cells without damage. The enzyme modification group's collection rate was 81% (residue rate 19%), while the grinding process group's collection rate was 56% (residue rate 44%)-an over 20% of collection rate difference. Thus we predicted a big difference in transference number after the process and wastage. In comparing ingredient changes in the enzyme modification group versus the grinding process group, the content of ${\beta}$-carotene (the carrot's main ingredient) showed a change in protection factor (PF) ($2.2{\pm}0.2$ PF, $1.4{\pm}0.4$ PF, respectively), total polyphenol content ($89{\pm}3.42{\mu}g/g$, $64{\pm}4.16{\mu}g/g$, respectively), and total flavonoid content ($68{\pm}2.73{\mu}g/g$, $41{\pm}3.26{\mu}g/g$, respectively). Thus we confirmed that nutrient destruction, due to cell membrane preservation, occurred less often in the enzyme modification process than the mechanical grinding process group. We also measured DPPH radical scavenging activity, hydroxyl radical scavenging activity, and nitrite scavenging activity. DPPH radical scavenging activity was $87{\pm}0.29%$ and $74{\pm}1.56%$ in the enzymatic modification group compared to the mechanical grinding process group, respectively. Hydroxyl radical scavenging activity was $44{\pm}0.49%$ and $32{\pm}0.48%$ in the enzymatic modification group compared to the mechanical grinding process group, respectively. Nitrite scavenging activity was $59{\pm}0.53%$ and $46{\pm}0.62%$ in the enzymatic modification group compared to the mechanical grinding process group, respectively. Our results show that cell membrane preservation, via the protopectinase enzyme process, decreases the loss of nutrients and still preserves inherent antioxidants.

• Korean journal of food and cookery science
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• v.9 no.4
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• pp.278-283
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• 1993

This study was attempted to investigate the effects of enzymatic modification of milk protein with protease on functional properties. The selected functional properties were solubility, emulsifying activity (EA), emulsion stability(ES), foam expansion(FE), and foam stability(FS). These properties were measu-red from pH 3.0 to pH 8.0. The proteases used in this study were iaolated from Meju(fermemted soybean) and had specific activity of 250 units/㎎ protein at pH 7.0, 1600 units of pretense was used for 1gr. of skim milk protein. Skim milk showed 30.5％ degree of hydrolysis for 1 hr. and 36.4% degree of hydrolysis for 3.5 hrs. of protease treatment at pH 7.0. Solubility of native skim milk, control, 1 hr. and 3.5 hrs. groups were 3.37, 3.64, 10.21, 14.34％o at pH 4.0 respcetively. The emulsifying activity of native skim milk, control, 1 hr. and 3.5 hrs. groups were 38.8,42.0,43.0,46.7ft at pH 4.0, respectively. Enzymatic modification resulted in the increase of solubility and emulsifying activity at pH 4.0. However at pH 5.0 emulsifying activity of 1 hr. and 3.5 hr. group were lower than native skim milk and control groups. 1 hr. protease treatment was found to be most effective way of increasing foam expansion at pH 4.0 to 6.0. It was supported that, protease treated skim milk can be used to improve solubility, emulsifying activity, foam expansion at acid pH. meju protease. skim milk, solubility, emulsion, foam.