• Title/Summary/Keyword: novel enzyme

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Antioxidant and Cytoprotective Effects of Socheongja and Socheong 2, Korean Black Seed Coat Soybean Varieties, against Hydrogen Peroxide-induced Oxidative Damage in HaCaT Human Skin Keratinocytes (HaCaT 인간 피부 각질세포에서 과산화수소 유도 산화 손상에 대한 소청자 및 소총2호의 항산화 및 세포보호 효능)

  • Choi, Eun Ok;Kwon, Da Hye;Hwang, Hye-Jin;Kim, Kook Jin;Lee, Dong Hee;Choi, Yung Hyun
    • Journal of Life Science
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    • v.28 no.4
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    • pp.454-464
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    • 2018
  • Black soybeans are used as food sources as well as for traditional medicines because they contain an abundance of natural phenolic compounds. In this study, total phenolic contents (TPCs) of Korean black seed coat soybean varieties Socheongja (SCJ), Socheong 2 (SC2) and Cheongja 2 (CJ2) as well as their antioxidant capacities were investigated. Among them, TPCs were abundantly present in the order of CJ2$H_2O_2$-stimulated HaCaT human keratinocytes. Our results revealed that treatment with SCJ and SC2 prior to $H_2O_2$ exposure significantly increases the viability of HaCaT cells, indicating that the exposure of HaCaT cells to SCJ and SC2 conferred a protective effect against oxidative stress. SCJ and SC2 also effectively inhibited $H_2O_2$-induced apoptotic cell death through the blocking of mitochondrial dysfunction. SCJ and SC2 also attenuated the phosphorylation of Histone H2AX. Furthermore, they effectively induced the levels of thioredoxin reductase (TrxR) 1, a potent antioxidant enzyme, which is associated with the induction of nuclear transcription factor erythroid-2-like factor 2 (Nrf2); however, the protective effects of SCJ and SC2 were significantly reversed by Auranofin, a TrxR inhibitor. These results indicate that they have protective activity through the blocking of cellular damage related to oxidative stress via the Nrf2 signaling pathway. In conclusion, our study indicated that SCJ and SC2 might potentially serve as novel agents for the treatment and prevention of skin disorders caused by oxidative stress.

Identification of a new marine bacterium Ruegeria sp. 50C-3 isolated from seawater of Uljin in Korea and production of thermostable enzymes (대한민국 울진 연안 해양에서 분리한 해양 미생물 Ruegeria sp. 50C-3의 동정 및 내열성 효소 생산)

  • Chi, Won-Jae;Kim, Jong-Hee;Park, Jae-Seon;Hong, Soon-Kwang
    • Korean Journal of Microbiology
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    • v.52 no.3
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    • pp.344-351
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    • 2016
  • A marine bacterium, designated as strain 50C-3, was isolated from a seawater sample collected from the East Sea of South Korea. The strain is a Gram-negative, aerobic, yellow colored polar-flagellated bacterium that grows at $20-50^{\circ}C$ and pH 5.5-8.5. Optimal growth occurred at $40-50^{\circ}C$, at pH 6.5-7.5, and in the presence of 2% (w/v) NaCl. Based on 16S rRNA gene sequence similarity, the isolate was considered to represent a member of the genus Ruegeria. The result of this analysis showed that strain 50C-3 shared 99.4% and 96.98% sequence similarity with Ruegeria intermedia CC-GIMAT-$2^T$ and Ruegeria lacuscaerulensis ITI-$1157^T$, respectively. Furthermore, strain 50C-3 showed clear differences from related strains in terms of several characteristics such as motility, carbon utilization, enzyme production, etc. The DNA G+C content was 66.7 mol%. Chemotaxonomic analysis indicated ubiquinone-10 (Q-10) as the predominant respiratory quinone. Based on phenotypic, chemotaxonomic, and phylogenetic characteristics, the isolate represents a novel variant of the Ruegeria intermedia CC-GIMAT-$2^T$, for which we named Ruegeria sp. 50C-3 (KCTC23890=DSM25519). Strain 50C-3 did not produce cellulase and agarase, but produced alkaline phosphatase, ${\alpha}$-galactosidase, and ${\beta}$-galactosidase. The three enzymes showed stable activities even at $50^{\circ}C$ and thus regarded as thermostable enzymes. Especially, the ${\beta}$-galactosidase activity enhanced by 1.9 times at $50^{\circ}C$ than that at $37^{\circ}C$, which may be very useful for industrial application.

Association Between the Polymorphism on Intron 5 of the Lipoprotein Lipase Gene and Carcass Traits in Hanwoo (Korean cattle) (한우 Lipoprotein Lipase 유전자 Intron 5번의 Polymorphism과 경제 형질과의 관련성 분석)

  • Lee, H.J.;Lee, S.H.;Cho, Y.M.;Yoon, H.B.;Jeon, B. K.;Oh, S.J.;Kwon, M.S.;Yoon, D.H.
    • Journal of Animal Science and Technology
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    • v.46 no.6
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    • pp.947-956
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    • 2004
  • The primary role of lipoprotein lipase(LPL) is the hydrolysis of triglycerides(TG) from the core of triglyceride-rich lipoproteins such as chylomicrons and very low density lipoproteins in plasma. Fatty acids liberated by LPL on capillary endothelial surfaces are available for tissues as energy sources especially in muscles or for storage in the form of TG in adipose tissues. Therefore, as the candidate gene related to the carcass traits of the beef cattle, we have directly sequenced the exon 5${\sim}$exon 6 region in the bovine LPL gene for discovery of single nucleotide polymorphism(SNP) with 24 unrelated Hanwoo(Korean cattle). Novel eight sequence variants were detected: three loci on exon 5, three on intron 5 and two on exon 6. All SNPs identified were strongly linked each other, and one hundred twenty eight Hanwoo samples were genotyped one SNP on intron 5 using PCR-restriction fragment length polymorphism method by digestion with Hae III restriction enzyme. The allele frequency of the polymorphism was 0.76 and 0.24. The effects of this polymorphism on the breeding values of the carcass weight, loin muscle area, back fat thickness and marbling score were analyzed using least square methods of SAS GLM. The marbling score of BB genotype was significantly higher than those of AA and AB genotypes(P<0.05). This result indicates that this polymorphism may be associated with the variation of marbling score. Further study is warranted to investigate the phenotypic association in Hanwoo.

Novel Method for Urinary 1-Hydroxypyrene Measurement Using Molecular Imprinting (분자주형을 이용한 요중 1-hydroxypyrene의 측정 방법 개발)

  • Yim, Dong-Hyuk;Moon, Sun-In;Choi, Young-Sook;Park, Hee-Jin;Kim, Dae-Seon;Yu, Seung-Do;Lee, Chul-Ho;Kim, Yong-Dae;Kim, Heon
    • Journal of Life Science
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    • v.21 no.4
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    • pp.549-553
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    • 2011
  • This study was performed to determine whether or not urinary 1-hydroxypyrene (1-OHP) levels can be accurately detected by our 1-OHP-detecting $TiO_2$-Bead-HPLC assay that we developed based on the molecular imprinting method. Our method showed a variation coefficient of 4.97% and a between-day variation coefficient of 4.43%, suggesting that this may be a very stable method. In addition, the recovery rate of 1-OHP from a mixture of 1-OHP and similar substances using our $TiO_2$-Bead-HPLC method was estimated to be 105.6%. The correlation coefficient between the conventional enzyme-HPLC method and this new method was 0.74 (p<0.01) when the urine samples were tested. Based on this result, it is conceivable that our method could be a useful technique for measuring urinary 1-OHP levels. Moreover, our method has some advantages of being easier and less expensive than the conventional method. The results of this study suggest that our method can facilitate the development of a urine 1-OHP sensor using $TiO_2$-coating beads and that development of beads by molecular imprinting can be applied to analysis of chemicals other than 1-OHP.

Biochemical Characterization of a Novel Thermostable Esterase from the Metagenome of Dokdo Islets Marine Sediment (독도 심해토 메타게놈 유래 신규 내열성 에스테라아제의 생화학적 특성규명)

  • Lee, Chang-Muk;Seo, Sohyeon;Kim, Su-Yeon;Song, Jaeeun;Sim, Joon-Soo;Hahn, Bum-Soo;Kim, Dong-Hern;Yoon, Sang-Hong
    • Microbiology and Biotechnology Letters
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    • v.45 no.1
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    • pp.63-70
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    • 2017
  • A functional screen of 60,672 fosmid metagenomic clones amplified from marine sediment obtained from the Dokdo islets in Korea identified the gene EstES1, whose product, EstES1, displayed lipolytic properties on tributyrin-supplemented media. EstES1 is a 576 amino acid protein with a predicted molecular weight of 59.4 kDa including 37 N-terminal leader amino acids. EstES1 exhibited the highest sequence similarity (44%) to a carboxylesterase found in Haliangium ochraceum DSM14365. Phylogenetic analysis indicated that EstES1 belongs to a currently uncharacterized family of lipases. Within the conserved domain, EstES1 retains the catalytic triad that consists of the consensus penta-peptide motif, GESAG. EstES1 demonstrated a broad substrate specificity toward the long acyl group of ethyl esters (C2-C12), and its optimal activity was recorded toward p-Nitrophenyl butyrate (C4) at pH 9.0 and $40^{\circ}C$ (specific activity of 255.4 U/mg). The enzyme remained stable in the ranges of $60-65^{\circ}C$ and pH 9.0-10.5 and in the presence of methanol, ethanol, isopropanol, and dimethyl sulfoxide. Therefore, EstES1 has potential for use in industrial applications involving high temperature, organic solvents, and/or alkaline conditions.

Promoter -202 A/C Polymorphism of Insulin-like Growth Factor Binding Protein-3 Gene and Non-small Cell Lung Cancer Risk (인슐린양 성장 인자 결합 단백-3 유전자 -202 좌위의 다형성에 따른 비소세포폐암의 위험도)

  • Moon, Jin Wook;Chang, Yoon Soo;Han, Chang Hoon;Kang, Shin Myung;Park, Moo Suk;Byun, Min Kwang;Chung, Wou Young;Park, Jae Jun;Yoo, Kyeong Nam;Shin, Ju Hye;Kim, Young Sam;Chang, Joon;Kim, Sung Kyu;Kim, Hee Jung;Kim, Se Kyu
    • Tuberculosis and Respiratory Diseases
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    • v.58 no.4
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    • pp.359-366
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    • 2005
  • Background : IGFBP-3 inhibits the mitogenic and anti-apoptotic activity of IGF by blocking the binding of IGF to its receptor. However, under certain circumstances, IGFBP-3 can enhance the activity of IGF by protecting IGF from its degradation. More than half of the interindividual variations in IGFBP-3 levels are known to be genetically determined by the polymorphism at -202 locus of IGFBP-3 gene. Method : We attempted to ascertain whether A-202C polymorphic variation of IGFBP-3 gene constitutes a risk factor for non-small cell lung cancer (NSCLC), using PCR-restriction fragment length polymorphism (RFLP). Our study included 104 NSCLC patients and 104 age-, gender-, and smoking status-matched control subjects. Result : In the 104 NSCLC subjects, the genotypic frequencies at the -202 site were as follows: AA = 67 (64.4%), AC = 35 (33.7%), and CC = 2 (1.9%). We did detect significant differences in the genotypic distribution between the NSCLC and the control subjects (p<0.05), and the NSCLC risk correlated significantly with AA genotype at the -202 locus (AA>AC>CC). Using CC genotype as a reference, the odds ratio (OR) for the subjects with AC genotype was 2.60 (95% CI: 0.89 - 8.60), and the OR associated with AA genotype was 5.89 (95% CI: 1.92 - 21.16). Conclusion : These results indicate that the dysregulation of IGF axis should now be considered as another important risk factor for NSCLC, and a potential target for novel antineoplastic therapies and/or preventative strategies in high-risk groups.

Somatic Cell Analysis and Cobalamin Responsiveness Study in Ten Korean Patients with Methylmalonic Aciduria (한국 메틸말로닌산혈증 환아 10례에서 Somatic Cell 분석과 cobalamin 반응성 연구)

  • Lim, Han Hyuk;Song, Wung Joo;Kim, Gu-Hwan;Watkins, David;Rosenblatt, David S.;Kim, Yoo-Mi;Chang, Mea Young;Kil, Hong Ryang;Kim, Sook Za
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.19 no.1
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    • pp.12-19
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    • 2019
  • Purpose: Isolated methylmalonic acidemia (MMA) is an autosomal recessive inherited disorder of propionate metabolism. There are two subtypes of MMUT gene defects. $Mut^0$ represents complete loss of methylmalonyl-CoA mutase (MCM) activity while mut- is associated with residual MCM activity, which can be stimulated by hydroxocobalamin (OHCbl) supplementation. The objective of this study is to investigate cobalamin responsiveness and mutations present in Korean MMA population. Methods: We evaluated 10 MMA patients using somatic cell complementation analysis on their fibroblasts to measure MCM activity and vitamin B12 responsiveness for the optimal treatment. MMUT gene was sequenced to identify the MMA mutations. Results: For all patients, the incorporation of $[^{14}C]-propionate$ was low, and there was no response to OHCbl. The incorporation of $[^{14}C]-methyltetrahydrofolate$ and $[^{57}Co]-CNCbl$ fell within the normal range. There was adequate synthesis of methylcobalamin while the synthesis of adenosylcobalamin was low. The complementation analysis showed all patients were $mut^0$. The sequence analysis identified 12 different MMUT mutations, including 2 novel mutations, p.Gln267Ter and p.Ile697Phe, were identified. All the patients in this study had neonatal onset of symptoms, belonged to $mut^0$ complementation class, and as a result, showed no cobalamin responsiveness. Conclusion: No Korean MMA patient showed cobalamin responsiveness.

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Protective Effect of the Ethyl Acetate-fraction of Methanol Extract of Ophiophogon japonicus on Amyloid beta Peptide-induced Cytotoxicity in PC12 Cells (소엽맥문동-에틸아세테이트 분획물의 아밀로이드 베타단백질-유발 세포독성에 대한 억제 효능)

  • Moon, Ja-Young;Kim, Eun-Sook;Choi, Soo-Jin;Kim, Jin-Ik;Choi, Nack-Shik;Lee, Kyoung;Park, Woo-Jin;Choi, Young-Whan
    • Journal of Life Science
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    • v.29 no.2
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    • pp.173-180
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    • 2019
  • Amyloid ${\beta}$-protein ($A{\beta}$) is the principal component of senile plaques characteristic of Alzheimer's disease (AD) and elicits a toxic effect on neurons in vitro and in vivo. Many environmental factors, including antioxidants and proteoglycans, modify $A{\beta}$ toxicity. It is worthwhile to isolate novel natural compounds that could prove therapeutic for patients with AD without causing detrimental side effects. In this study, we investigated the in vitro neuroprotective effects of the ethyl acetate fraction of methanol extract of Ophiophogon japonicas (OJEA fraction). We used an MTT reduction assay to detect protective effects of the OJEA fraction on $A{\beta}_{25-35}$-induced cytotoxicity to PC12 cells. We also used a cell-based ${\beta}$-secretase assay system to investigate the inhibitory effect of the OJEA fraction on ${\beta}$-secretase activity. In addition, we performed an in vitro lipid peroxidation assay to evaluate the protective effect of the OJEA fraction against oxidative stress induced by $A{\beta}_{25-35}$ in PC12 cells. The OJEA fraction had strong protective effects against $A{\beta}_{25-35}$-induced cytotoxicity to PC12 cells and was strongly inhibitory to ${\beta}$-secretase activity, which resulted in the attenuation of $A{\beta}$ generation. In addition, the OJEA fraction significantly decreased malondialdehyde (MDA) content, which is induced by the exposure of PC12 cells to $A{\beta}_{25-35}$. Our results suggested that the OJEA fraction contained active compounds exhibiting a neuroprotective effect on $A{\beta}$ toxicity.

A Novel Synthesized Tyrosinase Inhibitor, (E)-3-(4-hydroxybenzylidene) chroman-4-one (MHY1294) Inhibits α-MSH-induced Melanogenesis in B16F10 Melanoma Cells (신규 합성물질 (E)-3-(4-하이드록시벤질리딘)크로마논 유도체의 티로시나아제 효소활성 저해 및 멜라닌 생성 억제 효과)

  • Jeon, Hyeyoung;Lee, Seulah;Yang, Seonguk;Bang, EunJin;Ryu, Il Young;Park, Yujin;Jung, Hee Jin;Chung, Hae Young;Moon, Hyung Ryong;Lee, Jaewon
    • Journal of Life Science
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    • v.31 no.8
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    • pp.719-728
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    • 2021
  • Melanin pigments are abundantly distributed in mammalian skin, hair, eyes, and nervous system. Under normal physiological conditions, melanin protects the skin against various environmental stresses and acts as a physiological redox buffer to maintain homeostasis. However, abnormal melanin accumulation results in various hyperpigmentation conditions, such as chloasma, freckles, senile lentigo, and inflammatory pigmentation. Tyrosinase, a copper-containing enzyme, plays an important role in the regulation of the melanin pigment biosynthetic pathway. Although several whitening agents based on tyrosinase inhibition have been developed, their side effects, such as allergies, DNA damage, mutagenesis, and cytotoxicity of melanocytes, limit their applications. In this study, we synthesized 4-chromanone derivatives (MHY compounds) and investigated their ability to inhibit tyrosinase activity. Of these compounds, (E)-3-(4-hydroxybenzylidene)chroman-4-one (MHY1294) more potently inhibited the enzymatic activity of tyrosinase (IC50 = 5.1±0.86 μM) than kojic acid (14.3±1.43 μM), a representative tyrosinase inhibitor. In addition, MHY1294 showed competitive inhibitory action at the catalytic site of tyrosinase and had greater binding affinity at this site than kojic acid. Furthermore, MHY1294 effectively inhibited α-melanocyte stimulating hormone (α-MSH)-induced melanin synthesis and intracellular tyrosinase activity in B16F10 melanoma cells. The results of the present study indicate that MHY1294 may be considered as a candidate pharmacological agent and cosmetic whitening ingredient.

Evaluation of the Effects of Hangover-Releasing Agent Containing Vinegar Extract in Common Buckwheat and Tartary Buckwheat on Alcohol Metabolism and Hangover Improvement (일반메밀과 쓴메밀의 식초 추출물의 알코올 대사 및 숙취개선 효능 평가)

  • Su Jeong Kim;Hwang Bae Sohn;A Hyun Park;Jong Nam Lee;Su Hyoung Park;Jung Hwan Nam;Do Yeon Kim;Dong Chil Chang;Yul Ho Kim
    • Korean Journal of Plant Resources
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    • v.36 no.5
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    • pp.435-445
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    • 2023
  • The aim of this study was to explore the effects of vinegar extract from seed of common buckwheat (Fagopyrum esculentum Moench) and seed of tartary buckwheat (F. tataricum Gaertner) on acute ethanol-induced hangover in Sprague-Dawley rats. Vinegar extract from buckwheat is rich choline, quercetin and its glycoside, rutin known as flavonoid antioxidants. The test extract containing buckwheat was proven to alleviate hangovers through a significant reduction in the concentration of alcohol and acetaldehyde in the context of an alcohol-induced hangover model. Hepatic alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) activities were significantly higher in buckwheat vinegar-treated rats than in ethanol-treated rats. Moreover, tartary buckwheat vinegar upregulated antioxidant enzyme such as superoxide dismutase and Catalase activities in liver tissues. These results suggest that buckwheat vinegar extract could alleviate ethanol-induced hangover symptoms by elevating activities related to hepatic ethanol-metabolizing enzymes against ethanol induced metabolites, and in particular, tartary buckwheat should be further developed to be a novel anti-hangover material.