• Title/Summary/Keyword: Enzyme properties

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Whitening and inhibiting NF-κB-mediated inflammation properties of the biotransformed green ginseng berry of new cultivar K1, ginsenoside Rg2 enriched, on B16 and LPS-stimulated RAW 264.7 cells

  • Xu, Xing Yue;Yi, Eun Seob;Kang, Chang Ho;Liu, Ying;Lee, Yeong-Geun;Choi, Han Sol;Jang, Hyun Bin;Huo, Yue;Baek, Nam-In;Yang, Deok Chun;Kim, Yeon-Ju
    • Journal of Ginseng Research
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    • v.45 no.6
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    • pp.631-641
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    • 2021
  • Background: Main bioactive constituents and pharmacological functions of ripened red ginseng berry (Panax ginseng Meyer) have been frequently reported. Yet, the research gap targeting the beneficial activities of transformed green ginseng berries has not reported elsewhere. Methods: Ginsenosides of new green berry cultivar K-1 (GK-1) were identified by HPLC-QTOF/MS. Ginsenosides bioconversion in GK-1 by bgp1 enzyme was confirmed with HPLC and TLC. Then, mechanisms of GK-1 and β-glucosidase (bgp1) biotransformed GK-1 (BGK-1) were determined by Quantitative Reverse Transcription-Polymerase Chain Reaction and Western blot. Results: GK-1 possesses highest ginsenosides especially ginsenoside-Re amongst seven ginseng cultivars including (Chunpoong, Huangsuk, Kumpoong, K-1, Honkaejong, Gopoong, and Yunpoong). Ginseng root's biomass is not affected with the harvest of GK-1 at 3 weeks after flowering period. Then, Re is bioconverted into a promising pharmaceutical effect of Rg2 via bgp1. According to the results of cell assays, BGK-1 shows decrease of tyrosinase and melanin content in α-melanocyte-stimulating hormone challenged-murine melanoma B16 cells. BGK-1 which is comparatively more effective than GK-1 extract shows significant suppression of the nuclear factor (NF)-κB activation and inflammatory target genes, in LPS-stimulated RAW 264.7 cells. Conclusion: These results reported effective whitening and anti-inflammatory of BGK-1 as compared to GK-1.

Optimization of Submerged Culture Conditions for Protease Production and Its Enzymatic Properties (Protease 생산을 위한 최적 배양조건 및 생산된 Protease의 특성)

  • Cho, Hee-Yeon;Cho, Nam-Seok
    • Journal of the Korean Wood Science and Technology
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    • v.32 no.5
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    • pp.12-19
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    • 2004
  • This study was performed to investigate the optimum condition of protease production from submerged culture of oak mushroom (Lentinula edodes, Sanlim No. 5) and its enzymatic features. Among several combinations of media, the combination of wheat bran, corn flour, water and corn oil (WB+CF+W+ CO) yielded 84.8 U/g of maximum protease activity. This combination of ingredients, in spite of not being particularly protein-rich in comparison to the other media, allowed for good growth of the fungus and maximal protease production. Comparison of different growth medium liquids indicated that demineralized water afforded the best growth of the fungus and the highest protease activity. Acetate buffer and acidified water negatively affected The protease production peaked around 72 hr of incubation, and decreased thereafter. The molecular weights of produced protease were about 45,000 by Sephadex G-75 chromatography. The pH optimum for protease activity was 4, while maximal activity incubated at 37℃ for 1 hr was observed between pH 4~6. The optimum temperature of this protease was 55℃, and the enzyme was active over a broad temperature range (30~60℃), indicating that this protease would be suitable for a wide range of applications where. different pH and temperature are necessary, such as digestive aids, food industry, beer and tannery industries.

Evaluation of the Relationship between the Exposure Level to Mixed Hazardous Heavy Metals and Health Effects Using Factor Analysis (요인분석을 이용한 유해 중금속 복합 노출수준과 건강영향과의 관련성 평가)

  • Kim, Eunseop;Moon, Sun-In;Yim, Dong-Hyuk;Choi, Byung-Sun;Park, Jung-Duck;Eom, Sang-Yong;Kim, Yong-Dae;Kim, Heon
    • Journal of Environmental Health Sciences
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    • v.48 no.4
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    • pp.236-243
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    • 2022
  • Background: In the case of multiple exposures to different types of heavy metals, such as the conditions faced by residents living near a smelter, it would be preferable to group hazardous substances with similar characteristics rather than individually related substances and evaluate the effects of each group on the human body. Objectives: The purpose of this study is to evaluate the utility of factor analysis in the assessment of health effects caused by exposure to two or more hazardous substances with similar characteristics, such as in the case of residents living near a smelter. Methods: Heavy metal concentration data for 572 people living in the vicinity of the Janghang smelter area were grouped based on several subfactors according to their characteristics using factor analysis. Using these factor scores as an independent variable, multiple regression analysis was performed on health effect markers. Results: Through factor analysis, three subfactors were extracted. Factor 1 contained copper and zinc in serum and revealed a common characteristic of the enzyme co-factor in the human body. Factor 2 involved urinary cadmium and arsenic, which are harmful metals related to kidney damage. Factor 3 encompassed blood mercury and lead, which are classified as related to cardiovascular disease. As a result of multiple linear regression analysis, it was found that using the factor index derived through factor analysis as an independent variable is more advantageous in assessing the relevance to health effects than when analyzing the two heavy metals by including them in a single regression model. Conclusions: The results of this study suggest that regression analysis linked with factor analysis is a good alternative in that it can simultaneously identify the effects of heavy metals with similar properties while overcoming multicollinearity that may occur in environmental epidemiologic studies on exposure to various types of heavy metals.

Neuroprotective effect of Deodeok (Codonopsis lanceolata) bud extracts in H2O2-stimulated SH-SY5Y cells (더덕순 에탄올 추출물의 신경세포 보호 효과)

  • Hee Sun Yang;In Guk Hwang;Ae-jin Choi;Jeong-sook Choe
    • Journal of Nutrition and Health
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    • v.56 no.2
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    • pp.140-154
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    • 2023
  • Purpose: Deodeok (Codonopsis lanceolata) is generally used in conventional medicines and is considered to have remedial properties to cure several diseases. However, application of the C. lanceolata bud as a novel food ingredient has not been fully explored. Hydrogen peroxide (H2O2) is associated with the production of oxidative damage that results in mutagenesis, carcinogenesis, and cell death. This study examines the neuroprotective effect of C. lanceolate bud extracts (CLBE) on H2O2-stimulated apoptosis in SH-SY5Y cells. Methods: C. lanceolata bud of length 10 to 15 cm was collected and extracted using 70% ethanol. Cytotoxicity was evaluated by the EZ-cytox reagent, measurement of lactic dehydrogenase (LDH) release and reactive oxygen species (ROS). The morphological changes of the nuclei were determined using the Hoechst 33258 dye. Enzyme activities were analyzed using the caspase activity assay kit. Related protein expressions were quantified by the Western blot immunoassay in H2O2-stimulated SH-SY5Y cells. Results: Cell viability, LDH release and ROS generation, demonstrated neuroprotective effects of CLBE in H2O2-stimulated SH-SY5Y cells. The occurrence of apoptosis in H2O2-stimulated cells was confirmed by caspase activity, which was increased in H2O2-stimulated SH-SY5Y cells compared to the unexposed group. Pretreatment of CLBE was observed to inhibit the H2O2-stimulated apoptosis. In addition, exposure to CLBE resulted in increased expression of the Bcl-2 (B cell lymphoma 2) protein and decreased expression of the Bax (Bcl2 associated X) protein. Conclusion: This study shows that exposure to CLBE alleviates the H2O2-stimulated neuronal damage in SH-SY5Y cells. Our results indicate the potential application of CLBE in neurodegenerative disease therapy or prevention.

Anti-inflammatory Effect of Eleutherococcus senticosus Extracts from Gangwon-do by Plant parts or Solvents (강원도 가시오갈피의 식물 부위 또는 추출 용매 조건에 따른 항염증 효과)

  • Junkyu Park;Mina Boo;Soojin An;Sujin Shin;Jinbong Park;Ho-Young Choi;Kyungjin Lee
    • The Korea Journal of Herbology
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    • v.38 no.4
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    • pp.21-29
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    • 2023
  • Background : The purpose of this study was to investigate the anti-inflammatory properties of stems and leaves of Eleutherococcus senticosus (Rupr. & Maxim.) Maxim. (ES) from Gangwon-do. Methods and Results : Stems and leaves of ES were collected from two areas in Gangwon-do: Cheorwon-gun and Samcheok-si. Samples were extracted with water by using the pressurized liquid extraction method and with 70% prethanol A by using the heat reflux extraction method. The anti-inflammatory effects of ES were evaluated through the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide(MTT), lactate dehydrogenase(LDH) assay, nitric oxide(NO) assay, enzyme-linked immunosorbent assay(ELISA), and Western blot analysis in RAW 264.7 macrophages stimulated with lipopolysaccharide (LPS). 1) Results showed that ES leaf extractions were not cytotoxic at a concentration of up to 30 ㎍/㎖. The leaves of 70% prethanol A extractions of ES(30 ㎍/㎖) inhibited NO, interleukin-6(IL-6), and tumor necrosis factor-α(TNF-α) production and decreased the protein level of cyclooxygenase 2(COX-2). There was no significant change in the protein level of inducible nitric oxide synthase(iNOS). The stem extractions of ES did not exhibit anti-inflammatory effects. Conclusions : In this study, the leaves of 70% prethanol A extractions of ES demonstrated anti-inflammatory effect on RAW 264.7 macrophages. The 70% prethanol A extractions have a relatively higher anti-inflammatory effect on RAW 264.7 macrophages than water extractions.

Culture Conditions of E. coli Harboring Human O-Linked N-Acetyl-${\beta}$-Glucosaminidase Gene and Enzymatic Properties (사람의 O-linked-N-acetyl-${\beta}$-D-glucosaminidase 유전자를 함유한 대장균의 배양조건과 효소학적 특성)

  • 강대욱;조용권;서현효
    • Korean Journal of Microbiology
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    • v.40 no.2
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    • pp.147-153
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    • 2004
  • Protein modification by N-acetyl-${\beta}$-D-glucosamine (O-G1cNAc) on the hydroxyl groups of Ser or Thr ubiq-uitously occurs in eukaryotic cells and is involved in many cellular phenomena. The level of O-G1cNAc-mod-ified protein is regulated by OGT and O-GlcNAcase enzymes. We have tried to produce recombinant O-GlcNAcase in E. coli as an effort to establish in vitro screening system for modulators of O-GlcNAcase. The culture conditions for improvement of O-GlcNAcase productivity, were as follows: induction temperature, $30^{\circ}C$; the concentration of L-arabinose, 0.02% and induction time, 5 hr. Under these culture conditions, E. coli cells containing O-GlcNAcase gene had no enzyme activity until up to 3 hr culture. However, O-GlcNAcase activity dramatically increased from 3 to 5 hr culture. It almost maintained the same level after 5 hr culture. Western blot analysis verified the amount of expressed O-GlcNAcase increased with culture time, being con-sistent with activity data. The optimal reaction condition determined in this study was as follows: protein quan-tity, $5{\mu}g$; reaction time, 30 min; reaction temperature, $45^{\circ}C$; substrate concentration, 2 mM; reaction pH, 6.5. Methanol had little effect on O-GlcNAcase activity and 90% of activity were retained at 10%. Only 15% resid-ual activity were detected at 5% of chloroform.

Enhanced Antioxidative Potential by Silymarin Treatment through the Inductionof Nrf2/MAPK Mediated HO-1 Signaling Pathway in RAW 264.7 Cells (RAW 264.7 세포에서 Nrf2/MAPK 의 활성을 통한 HO-1 과발현에 의한 silymarin의 항산화 효과)

  • Hyun-Seo Yoon;Hyun An;Chung Mu Park
    • Journal of Life Science
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    • v.33 no.10
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    • pp.776-782
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    • 2023
  • Silymarin, which is derived from dried Silybum marianum (milk thistle) seeds and fruits, possesses various beneficial properties, such as hepatoprotective, antioxidative, anti-inflammatory, and anticancer activity. This research aimed to explore the antioxidative activity of silymarin against oxidative stress and understand its molecular mechanism in RAW 264.7 cells. The study employed cell viability and reactive oxygen species (ROS) formation assays and western blot analysis. The results demonstrated that silymarin effectively reduced intracellular ROS levels induced by lipopolysaccharide (LPS) in a dose-dependent manner without causing any cytotoxic effects. Moreover, silymarin treatment significantly upregulated the expression of heme oxygenase (HO)-1, a phase II enzyme known for its potent antioxidative activity. Additionally, silymarin treatment significantly induced the expression of nuclear factor-erythroid 2 p45-related factor (Nrf) 2, a transcription factor responsible for regulating antioxidative enzymes, which was consistent with the upregulated HO-1 expression. To investigate the involvement of key signaling pathways in maintaining cellular redox homeostasis against oxidative stress, the phosphorylation status of mitogen-activated protein kinase (MAPK) and phosphoinositide 3-kinase (PI3K) was estimated by western blot analysis. The results showed that silymarin potently induced HO-1 expression, which was mediated by the phosphorylation of p38 MAPK. To further validate the antioxidative potential of silymarin-induced HO-1 expression, tert-butyl hydroperoxide (t-BHP)-induced oxidative damage was employed and attenuated by silymarin treatment, as identified by a selective inhibitor for each signaling molecule. In conclusion, silymarin robustly enhanced antioxidative activity by inducing HO-1 via the Nrf2/p38 MAPK signaling pathway in RAW 264.7 cells.

Recent Progress in Colorimetric Assays Using the Absorption of Plasmonic Gold Nanoparticles (플라즈모닉 금 나노입자의 흡광 특성을 활용한 생화학적 비색 분석법 연구 동향)

  • Bong-Geun Kim;Sang Bin Yoon;Sukyeong Hwang;Hyon Bin Na
    • Applied Chemistry for Engineering
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    • v.35 no.2
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    • pp.67-78
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    • 2024
  • Light absorption has potential as a signal in biochemical analyses due to its simplicity in measurement and interpretational clarity. Among substances that generate absorption signals, gold nanoparticles possess advantages such as chemical stability, biological compatibility, and unique optical properties from the localized surface plasmon resonance (LSPR) in the visible light range. They also exhibit versatility compared to other colorimetric substances effective only for specific target molecules, as they easily conjugate with various detection active substances like antibodies and aptamers. Particularly due to advantages such as low cost, ease of particle synthesis, and high environmental stability compared to enzyme-based colorimetric methods, gold nanoparticles are extensively researched as signal substances in colorimetric assays. This review summarizes various strategies utilizing gold nanoparticles as absorption signal substances, focusing on recent research. Based on the characteristics of gold nanoparticles, where the optical property is influenced by particle morphology, literature is classified and reviewed based on strategies controlling the shape of gold nanoparticles during signal generation. Through this, it is observed that gold nanoparticles, which have been used as absorption signal substances, continue to be actively researched, affirming their potential for broad and continuous improvement in the future.

Effects of Provitamin D on Skin Pigmentation and Immunity (프로 비타민 D 의 피부색 조절 및 면역 효능)

  • Hyunjung Choi;Daejin Min;Eun-Jeong Choi;Seung-Han Park;Hyoung-June Kim;Won-Seok Park
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.50 no.2
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    • pp.153-161
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    • 2024
  • Vitamin D is a fat-soluble vitamin that is mainly produced in the skin by UV rays. Along with melatonin, it is a representative chronobiotic substance, and the skin plays an important role in distinguishing between day and night. However, vitamin D cannot be used directly in cosmetics because it is a vitamin that acts as a coenzyme and plays a hormonal role in regulating the expression of various types of genes. Therefore, it was to investigate the skin efficacy of provitamin D (7-dehydrocholesterol), a vitamin D precursor that can be used in cosmetics. Our findings reveal that pro vitamin D can effectively inhibit the expression of tyrosinase, the melanin-producing enzyme, thereby attenuating melanin synthesis. This skin tone regulatory effect has been corroborated in vitro using artificial skin models. Additionally, pro vitamin D demonstrated anti-inflammatory properties by suppressing the expression of TNFa and, upon conversion to vitamin D through UV exposure, it was observed to induce the production of the antimicrobial peptide CAMP (LL-37). The inhibitory effect of pro vitamin D on melanin production appears to be a result of it reducing the UV absorption capacity of melanin, thereby inducing the conversion of pro D to vitamin D. Utilizing pro vitamin D in cosmetic formulations could not only modulate skin tone and enhance skin immunity but also expect to contribute to other cutaneous benefits as anti-aging and barrier function improvement with everyday UV exposure. This multifaceted efficacy positions pro vitamin D as a promising ingredient in advancing the formulation of skin care products.

Discovery of a Novel Cellobiose Dehydrogenase from Cellulomonas palmilytica EW123 and Its Sugar Acids Production

  • Ake-kavitch Siriatcharanon;Sawannee Sutheeworapong;Sirilak Baramee;Rattiya Waeonukul;Patthra Pason;Akihiko Kosugi;Ayaka Uke;Khanok Ratanakhanokchai;Chakrit Tachaapaikoon
    • Journal of Microbiology and Biotechnology
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    • v.34 no.2
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    • pp.457-466
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    • 2024
  • Cellobiose dehydrogenases (CDHs) are a group of enzymes belonging to the hemoflavoenzyme group, which are mostly found in fungi. They play an important role in the production of acid sugar. In this research, CDH annotated from the actinobacterium Cellulomonas palmilytica EW123 (CpCDH) was cloned and characterized. The CpCDH exhibited a domain architecture resembling class-I CDH found in Basidiomycota. The cytochrome c and flavin-containing dehydrogenase domains in CpCDH showed an extra-long evolutionary distance compared to fungal CDH. The amino acid sequence of CpCDH revealed conservative catalytic amino acids and a distinct flavin adenine dinucleotide region specific to CDH, setting it apart from closely related sequences. The physicochemical properties of CpCDH displayed optimal pH conditions similar to those of CDHs but differed in terms of optimal temperature. The CpCDH displayed excellent enzymatic activity at low temperatures (below 30℃), unlike other CDHs. Moreover, CpCDH showed the highest substrate specificity for disaccharides such as cellobiose and lactose, which contain a glucose molecule at the non-reducing end. The catalytic efficiency of CpCDH for cellobiose and lactose were 2.05 × 105 and 9.06 × 104 (M-1 s-1), respectively. The result from the Fourier-transform infrared spectroscopy (FT-IR) spectra confirmed the presence of cellobionic and lactobionic acids as the oxidative products of CpCDH. This study establishes CpCDH as a novel and attractive bacterial CDH, representing the first report of its kind in the Cellulomonas genus.