• Applied Biological Chemistry
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• v.46 no.3
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• pp.171-175
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• 2003

The putative endochitinase gene, yheB of Escherichia coli K-12 is not expressed under lab culture conditions. The endochitinase gene was amplified by PCR and subcloned into pET28c vector and pQE9 vector, respectively. The endochitinase produced in E. coli harboring pET28c containing yheB or pQE9 vector containing yheE was partly released into the growth medium. The overproduced endochitinase was partially purified by His affinity column chromatography and DE-52 column chromatography. The apparent molecular weight of the endochitinase determined by SDS-polyacrylamide gel electrophoresis was about 97,000. The purified E. coli endochitinase showed maximal chitinolytic activity at pH 6 and $40^{\circ}C$.

• Journal of Korean Medicine for Obesity Research
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• v.3 no.1
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• pp.7-16
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• 2003

Objectives : The lipolytic effects of catecholamines in adipose tissue are mediated by members of adrenergic receptors. This study was conducted to examine the effects of ${\beta}2-AR$ Gln27Glu (Q27E) polymorphism on obesity indices and risk among obesity clinic patients. Methods : 532 subjects, 38 men and 494 women, who attended a weight loss program in a local obesity clinic were analyzed. Height, weight, BMI, WHR and obesity index were measured or calculated. Body composition was measured by bio-impedance analysis. Genotype of ${\beta}2-AR$ polymorphism in codon 27 was analyzed by PCR-RFLP method. Serum concentrations of fasting glucose, total and HDL cholesterol, and triglyceride were determined by autobiochemical analyzer. Results: The Genotype distributions of ${\beta}2-AR$ gene were QQ type 81.3%, QE type 17.9% and EE type 8%. Therefore, the frequency of E allele of ${\beta}2-AR$ gene was 0.170 in the total subjects. The frequency of the ${\beta}2-AR$ variant genotype(QE+EE) was significantly higher in obese group($BMI{\geqq}25$) compared with non-obese group(p=0.027). Weight was significantly higher in variant(QE+EE) type compared with normal(QQ) type in total subjects(p=0.001), male(p=0.022) and female(p=0.013). BMI, obesity index and WHR were also significantly higher in QE+EE type. Body fat man was significantly higher in QE+EE type in total subjects(p=0.005) and female(p=0.027). When forward stepwise regression analysis was used to create a model of risk predictors of obesity($BMI{\geqq}25$), QE+EE type of ${\beta}2-AR$ gene was found to be a significant risk factor for obesity (p=0.042, ORs 1.597). Conclusions: QE+EE genotype of ${\beta}2-AR$ was associated with increased obesity indices and might be a significant risk factor for obesity.

• Journal of the Korean Society of Food Science and Nutrition
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• v.44 no.4
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• pp.557-563
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• 2015

To observe the functionality of Fuji apples, this study compared and analyzed the general and anti-oxidative components of apples based on production region. This study found that DPPH radical scavenging activities among parts of apple from the Chungju region were 82.84% in peels, 26.98% in peel-flesh, and 18.89% in apple flesh, and these values were lower than those from other regions (P<0.01). Antioxidative was 48.64% in the apple core, which was higher than those in peel-flesh and apple flesh. ABTS radical scavenging activity was highest (79.80%) in peels of apples from the Andong region, whereas values in peel-flesh and apple flesh were highest in apples from the Yesan region (P<0.01). For antioxidative activities in the apple core, apples from the Chungju region showed more than twice the value (52.34%) of other regions. Phenol contents in peels were significantly high [12.03 mg gallic acid equivalent (GAE)/g] in apples from the Muju region, whereas phenol contents in peel-flesh were high (6.01 mg GAE/g) in those from the Andong region. Antioxidative activity in apple flesh was significantly high (5.57 mg GAE/g) in apples from the Yesan region. For antioxidative activities in the apple core, apples from Chungju region showed a relatively high value (6.53 mg GAE/g) (P<0.01), although values were low in apple peel, peel-flesh, and apple flesh. For the combined amount of flavonoids, values in apples from the Yesan region were high in apple peel, peel-flesh, and apple core [56.23 mg quercetin equivalent (QE)/g (P<0.01), 4.05 mg QE/g (P<0.05), and 4.00 mg QE/g (P<0.01), respectively], whereas flavonoid content in apples from the Andong region was high in apple flesh [4.35 mg QE/g (P<0.01)]. The results show that anti-oxidative activities were relatively higher in apple peel than flesh.

• Korean Journal of Veterinary Research
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• v.46 no.1
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• pp.13-19
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• 2006

Generally known psittacosis or ornithosis is a disease of birds caused by the bacterium Chlamydia psittaci. Humans are accidential hosts and are most commonly infected from avian sources. It raises hepatitis or neurosis. As major outer membrane protein (MOMP) of Chlamydia psittaci has been known to play a role in the avoidance of host immune defenses, research on developing a Chlamydia vaccine has focused on the MOMP. In this study, the gene encoding the major outer membrane protein (MOMP) of the Chlamydia psittaci strain 6BC was cloned and expressed in Escherichia coli strain M-15. The recombinant DNA was cloned by fusion prokaryotic expression vector pQE30-GFPII. Expression of the recombinant protein was performed in E. coli and was induced by IPTG. The size of expressed recombinant protein is 74.220 kDa (MOMP, 43.260 kDa; GFP expression region, 30 kDa; $6{\times}His$ tag, 960Da). This protein was purified by using his-tagging-inclusion body. Recombinant protein was reconfirmed through ELISA test and western blot with antibody against pQE30-GFPII. It will be useful antibody development.

• The Journal of Korean Society of Virology
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• v.28 no.3
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• pp.215-224
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• 1998

Cloning, sequencing and expressing in E. coli of the thymidine kinase (TK) gene of Herpes simplex virus type-1 (HSV-1) strain F was investigated. The TK gene, located in the BamHI 3.74 kb DNA fragment of the plasmid pHLA-12, was amplified by polymerase chain reaction (PCR). The 1,131 kb PCR product was cloned into the BamHI and EcoRI sites of pBacPAK9 plasmid and then named pBac-TK recombinant. The TK gene was subcloned into the BamHI and BglII sites of pQE-30, and named pQE-TK recombinant. The nucleotide sequence of the 1,131 kb TK gene was determined, and the GC content was 65.13%. There were deduced 367 amino acid residues with a total molecular weight of 43 kDa. The weight was confirmed by the protein produced by E. coli M15/pQE-TK on the SDS-PAGE and Western blot. The production of the TK protein in the IPTG induced cells was measured over 4 h. At the end of 1, 2 and 3 h the level increased by 146, 204 and 242%, respectively. The amount of the protein at the highest fraction purified with Ni-NTA resin chromatography was $0.68\;{\mu}g$ per ml. The soluble state TK protein was present in the cytoplasm. In these results the F strain was different in base sequence and amino acid sequence from that of the CL101 strain, which caused difference in their strains.

• Biotechnology and Bioprocess Engineering:BBE
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• v.2 no.2
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• pp.86-89
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• 1997

Epidermal growth factor(EGF) known as a urgastrone is a powerful mitogen with a wide variety of possibilities for medical usages. A mature EGF coding region was isolated from human prepro-EGF sequence by a conventional PCR and cloned into pQE vector in which the gene product was supposed to be expressed with 6$\times$His tag for the subsequent purification. The recombinant mature EGF was expressed in M15[Rep4], an Escherichia coli host strain, in amount of 30-40% of total proteins pressent in E. coli extract by the addition of isopropylthio-$\beta$-galactopyranoside (IPTG). The recombinant EGF purified using a Ni2+-NTA affinity colume chromatography was active in its ability to induce phosphorylation on tyrosine residues of several substrate proteins when murine NH3T3 and human MRC-5 fibroblast cells were stimulated with it. This work may provide the basic technology and information for the production of recombinant EGF.

• Korean Journal of Microbiology
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• v.38 no.4
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• pp.241-246
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• 2002

A gene coding for endo-$\beta$-1,3-1,4-glucanase of Bacillus circulans was subcloned into Escherichia coli Ml5 using pQE30 as an expression vector. Endo-$\beta$-1,3-1,4-glucanase produced by the recombinant expression plas-mid pLQ43 was intactly purified to a single protein through a nickel-nitrilotriacetic acid (Ni-NTA) metal-affinity chromatography method. The molecular mass of the purified enzyme was estimated to be 28 kDa by SDS-PAGE. The optimum pH and temperature of the enzyme activity were pH 6.8 and $60^{\circ}C$, respectively. This enzyme was fairly stable in the pH ranging 5.5~7.5 and at the temperatures lower than $55^{\circ}C$. The enzyme appeared to be sensitive to most of the metal ions, especially to $Hg^{2＋$, and also to methanol, ethanol, isopropanol or 1-butanol at a concentration of 10%(v/v).

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.20
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• pp.8685-8688
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• 2014

The objective of the present study was to investigate cloning, expression, and functions of the recombinant protein, Siva1. Siva1 gene was synthesized by RT-PCR from HCT116 cells. Plasmids were cleaved with the restriction endonuclease, BamH1/Sal1 and products were connected to pQE30, which underwent cleavage by BamH1/Sal1. The recombinant plasmid, pQE30-Siva1, was identified after digestion with restriction endonucleases followed by transformation into E. coli M15. Expression of Siva1 was induced by IPTG and identified by SDS-PAGE following purification with affinity chromatography. The results showed that size of Siva1 was 12 kDa, consistent with the molecular weight of the His-Siva1 fusion protein. Functional test demonstrated that Siva1 significantly inhibited the invasion and migration of HCT116 cells. It may thus find clinical application for control of cancers.

• Food Engineering Progress
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• v.21 no.3
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• pp.249-255
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• 2017

Caesalpinia sappan L. is an oriental medicinal plant distributed in the Asia Pacific region including India, Malaysia, and China. The dried heartwood of Caesalpinia sappan has been traditionally used as an analgesic and anti-inflammatory drug. In this study, the effects of extract methods of C. sappan on contents of total polyphenols and flavonoids, antioxidant activity, and cytotoxic activity were evaluated. As a result, hot water extract from C. sappan (CSWE) significantly exhibited contents of total polyphenols and flavonoids (22.6 mg GAE/g and 14.5 mg QE/g) higher than 70% ethanol extract (CSEE) (17.6 mg GAE/g and 13.2 mg QE/g). However, CSEE showed greater antioxidant activity than CSWE in both DPPH and ABTS. Also, the cytotoxicity of C. sappan against three kinds of cancer cell lines was higher in CSEE than in CSWE. These results show that ethanol extract is a better extract method than hot water method to maintain antioxidant and anti-cancer activities.

• Culinary science and hospitality research
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• v.21 no.6
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• pp.120-132
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• 2015

The purpose of this study was to provide preliminary information for the utilization extension of Chinese artichoke(Stachys sieboldii Miq) as a functional food material. The effects of the addition of Chinese artichoke powder(0, 3, 6, 9, and 12%) in white bread formulation on phenolics content and antioxidant properties, and sensory analysis(seven-point hedonic test) were examined. The contents of total polyphenols(TPC), flavonoids (TFC), and tannins(TTC) in Chinese artichoke powder were $139.09{\pm}1.97mg\;GAE/g\;dw$, $74.33{\pm}2.69mg\;QE/g\;dw$, and $40.41{\pm}2.54 mg\;TAE/g\;dw$, respectively. As the amount of Chinese artichoke powder increased, the phenolics contents also significantly increased(p<0.001, p<0.001, and p<0.001 on TPC, TFC, and TTC, respectively), the highest TPC($104.27{\pm}0.13mg\;GAE/g\;dw$), TFC($71.03{\pm}1.75mg\;QE/g\;dw$), and TTC($8.76{\pm}0.12mg\;TAE/g\;dw$) were achieved in the white bread having the highest percentage of Chinese artichoke powder(12%). The $IC_{50}$ values of Chinese artichoke extract for 1,1-diphenyl-2-picrylhydrazyl(DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid(ABTS) radical scavenging activities were 1.42 mg/mL and 1.57 mg/mL, respectively. Scavenging activities of DPPH and ABTS radicals of white bread were significantly increased according to the levels of added Chinese artichoke powder(p<0.001 and p<0.001, respectively). In the acceptance test, the white bread containing 9% Chinese artichoke powder was ranked significantly higher than the other groups according to all sensory parameters such as appearance, flavor, taste, texture, and the overall acceptability. Overall, Chinese artichoke enhanced white bread could be developed as an antioxidant-enriched bread with good sensorial properties.