• Title/Summary/Keyword: degradation enzyme

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The Analysis of Whitening Effects on Extracts from Ginkgo (Ginkgo biloba L.) Seeds (은행나무 종자 추출물의 미백효능 분석)

  • Choi, Eun-Young;Jang, Young-Ah
    • Journal of the Korean Applied Science and Technology
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    • v.38 no.5
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    • pp.1229-1240
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    • 2021
  • Ginkgo (Ginkgo biloba L.) seeds, called 'Baekqwa', were extracted from 70% ethanol to investigate the whitening effect and to confirm the application potential as a cosmetic material. Ginkgo seed ethanol extracts (GBE) were treated with B16F10 melanoma cells, and melanin contents and tyrosinase, which is the main enzyme concerning the synthesis process of melanin, inhibitory activity were confirmed. As a result, there were inhibited in a concentration-dependent manner, and GBE also significantly reduced the protein expression and mRNA levels of tyrosinase, tyrosinase-related protein-1, -2 (TRP-1, -2), and their upstream transcription factor, microphthalmia-associated transcription factor (MITF). These results suggest that GBE could be used as an effective whitening agent that has an inhibitory effect on melanin production by regulating the expression and degradation of MITF in melanocytes.

Bioconversion of Lignocellulosic Materials with the Contribution of a Multifunctional GH78 Glycoside Hydrolase from Xylaria polymorpha to Release Aromatic Fragments and Carbohydrates

  • Liers, Christiane;Ullrich, Rene;Kellner, Harald;Chi, Do Huu;Quynh, Dang Thu;Luyen, Nguyen Dinh;Huong, Le Mai;Hofrichter, Martin;Nghi, Do Huu
    • Journal of Microbiology and Biotechnology
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    • v.31 no.10
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    • pp.1438-1445
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    • 2021
  • A bifunctional glycoside hydrolase GH78 from the ascomycete Xylaria polymorpha (XpoGH78) possesses catalytic versatility towards both glycosides and esters, which may be advantageous for the efficient degradation of the plant cell-wall complex that contains both diverse sugar residues and esterified structures. The contribution of XpoGH78 to the conversion of lignocellulosic materials without any chemical pretreatment to release the water-soluble aromatic fragments, carbohydrates, and methanol was studied. The disintegrating effect of enzymatic lignocellulose treatment can be significantly improved by using different kinds of hydrolases and phenoloxidases. The considerable changes in low (3 kDa), medium (30 kDa), and high (> 200 kDa) aromatic fragments were observed after the treatment with XpoGH78 alone or with this potent cocktail. Synergistic conversion of rape straw also resulted in a release of 17.3 mg of total carbohydrates (e.g., arabinose, galactose, glucose, mannose, xylose) per gram of substrate after incubating for 72 h. Moreover, the treatment of rape straw with XpoGH78 led to a marginal methanol release of approximately 17 ㎍/g and improved to 270 ㎍/g by cooperation with the above accessory enzymes. In the case of beech wood conversion, the combined catalysis by XpoGH78 and laccase caused an effect comparable with that of fungal strain X. polymorpha in woody cultures concerning the liberation of aromatic lignocellulose fragments.

Comparison of nitrogen transformation dynamics in non-irradiated and irradiated alfalfa and red clover during ensiling

  • Dong, Zhihao;Li, Junfeng;Chen, Lei;Yuan, Xianjun;Shao, Tao
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.10
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    • pp.1521-1527
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    • 2019
  • Objective: To study the contribution of plant enzyme and microbial activities on protein degradation in silage, this study evaluated the nitrogen transformation dynamics during ensiling of non- and irradiated alfalfa (Medicago sativa L.) and red clover (Trifolium pratense L.). Methods: Alfalfa and red clover silages were prepared and equally divided into two groups. One group was exposed to ${\gamma}$-irradiation at a recommended dosage (25 Gky). Therefore, four types of silages were produced: i) non-irradiated alfalfa silage; ii) irradiated alfalfa silage; iii) non-irradiated red clover silage; and iv) irradiated red clover silage. These silages were opened for fermentation quality and nitrogen components analyses after 1, 4, 8, and 30 days, respectively. Results: The ${\gamma}$-irradiation successfully suppressed microbial activity, indicated by high pH and no apparent increases in fermentation end products in irradiated silages. All nitrogen components, except for peptide-N, increased throughout the ensiling process. Proteolysis less occurred in red clover silages compared with alfalfa silages, indicated by smaller (p<0.05) increment in peptide-N and free amino acid N (FAA-N) during early stage of ensiling. The ${\gamma}$-irradiation treatment increased (p<0.05) peptide-N and FAA-N in alfalfa silage at day 1, whereas not in red clover silage; these two nitrogen components were higher (p<0.05) between day 4 and day 30 in non-irradiated silages than the irradiated silages. The ammonia nitrogen and non-protein nitrogen were highest in non-irradiated alfalfa silage and lowest in irradiated red clover silage after ensiling. Conclusion: The result of this study indicate that red clover and alfalfa are two forages varying in their nitrogen transformation patterns, especially during early stages of ensiling. Microbial activity plays a certain role in the proteolysis and seems little affected by the presence of polyphenol oxidase in red clover compared with alfalfaa.

Bioconversion enhances anti-oxidant and anti-inflammation activities of different parts of the Mulberry Tree (Morus alba L.), especially the leaf (Mori Folium)

  • Chon, So-Hyun;Kim, Min-A;Lee, Han-Saem;Park, Jeong-Eun;Lim, Yu-Mi;Kim, Eun-Jeong;Son, Eun-Kyung;Kim, Sang-Jun;So, Jai-Hyun
    • Journal of Applied Biological Chemistry
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    • v.62 no.2
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    • pp.111-122
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    • 2019
  • The mulberry tree (Morus alba L.) has been traditionally used in Chinese medicine to treat inflammatory diseases. We investigated the effects of bioconversion on different components of the mulberry tree, and determined changes in the physiological activities. Ethyl acetate-soluble fractions of five different segments (fruit, Mori Fructus; leaf, Mori Folium; twig, Mori Ramulus; root, Mori Cortex; and mistletoe, Loranthi Ramulus) of the mulberry tree show enhanced anti-oxidant effects in the 2,2-diphenyl-1-picrylhydrazyl, and 2,2'-azinobis-(3-ethylvenzothiazoline-6-sulfonic acid) assays, and enhanced anti-inflammatory effects of lipopolysaccharide (LPS)-stimulated nitric oxide (NO) production in RAW 264.7 macrophages, after being treated with a crude enzyme extract from Aspergillus kawachii, in the following order of activity: Mori Folium>Mori Cortex>Mori Ramulus>Mori Fructus>Loranthi Ramulus. Ethyl acetate- soluble fraction of mulberry leaves (Mori Folium) that underwent bioconversion was most effective, and was devoid of any cytotoxicity. The fraction was also effective against mRNA expression of LPS-induced pro-inflammatory cytokines, such as inducible nitric oxide synthase, cyclooxygenase-2, tumor necrosis $factor-{\alpha}$, $interleukin-1{\beta}$, and interleukin-6. In addition, the fraction was effective in LPS-induced phosphorylation of mitogen-activated protein kinases and IKK, and $I{\kappa}B$ degradation, followed by translocation of the nuclear $factor-{\kappa}B$ from the cytoplasm to the nucleus. Thus, bioconversion increased the anti-oxidative and anti-inflammatory activities of the mulberry leaf.

Effect of Medicinal Plant Extracts on the Ethanol-Metabolizing Enzyme Activities (약용식물 추출물의 에탄올대사 효소활성에 미치는 영향)

  • Do, Jaeho;Gwak, Jungwon;Lee, Sunjeong;Rho, Jung Jin;Lee, Kwangseung;Kim, Dong Chung
    • Food Engineering Progress
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    • v.21 no.3
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    • pp.286-291
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    • 2017
  • This study was conducted to certify the effect of aqueous extracts from fifty medicinal plants on the activities of alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) in vitro. Each aqueous extract was prepared by combining one-part medicinal plants with twenty-parts distilled water at $80^{\circ}C$ for 8 h. Among the fifty medicinal plants, Allium sativum L. and Cinnamomum cassia Presl were regarded as an effective anti-hangover substance. Allium sativum L. extract increased ALDH activity more than 2 times compared with ADH activity, enhancing the acetaldehyde degradation. Cinnamomum cassia Presl extract dramatically inhibited ADH activity compared with ALDH activity, thus potently decreasing the acetaldehyde formation. ADH and ALDH activities were proportionally inhibited according to the increased concentration of Cinnamomum cassia Presl extract. The aqueous extract of Cinnamomum cassia Presl at a concentration of $45.33{\mu}g/mL$ inhibited ADH activity by 52.8% and ALDH activity by 11.0%.

ACOX1 destabilizes p73 to suppress intrinsic apoptosis pathway and regulates sensitivity to doxorubicin in lymphoma cells

  • Zheng, Fei-Meng;Chen, Wang-Bing;Qin, Tao;Lv, Li-Na;Feng, Bi;Lu, Yan-Ling;Li, Zuo-Quan;Wang, Xiao-Chao;Tao, Li-Ju;Li, Hong-Wen;Li, Shu-You
    • BMB Reports
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    • v.52 no.9
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    • pp.566-571
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    • 2019
  • Lymphoma is one of the most curable types of cancer. However, drug resistance is the main challenge faced in lymphoma treatment. Peroxisomal acyl-CoA oxidase 1 (ACOX1) is the rate-limiting enzyme in fatty acid ${\beta}$-oxidation. Deregulation of ACOX1 has been linked to peroxisomal disorders and carcinogenesis in the liver. Currently, there is no information about the function of ACOX1 in lymphoma. In this study, we found that upregulation of ACOX1 promoted proliferation in lymphoma cells, while downregulation of ACOX1 inhibited proliferation and induced apoptosis. Additionally, overexpression of ACOX1 increased resistance to doxorubicin, while suppression of ACOX1 expression markedly potentiated doxorubicin-induced apoptosis. Interestingly, downregulation of ACOX1 promoted mitochondrial location of Bad, reduced mitochondrial membrane potential and provoked apoptosis by activating caspase-9 and caspase-3 related apoptotic pathway. Overexpression of ACOX1 alleviated doxorubicin-induced activation of caspase-9 and caspase-3 and decrease of mitochondrial membrane potential. Importantly, downregulation of ACOX1 increased p73, but not p53, expression. p73 expression was critical for apoptosis induction induced by ACOX1 downregulation. Also, overexpression of ACOX1 significantly reduced stability of p73 protein thereby reducing p73 expression. Thus, our study indicated that suppression of ACOX1 could be a novel and effective approach for treatment of lymphoma.

Wheat phytase can alleviate the cellular toxic and inflammatory effects of lipopolysaccharide

  • An, Jeongmin;Cho, Jaiesoon
    • Journal of Animal Science and Technology
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    • v.63 no.1
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    • pp.114-124
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    • 2021
  • The objective of this study was to characterize the enzymatic hydrolysis of lipopolysaccharide (LPS) by wheat phytase and to investigate the effects of wheat phytase-treated LPS on in vitro toxicity, cell viability and release of a pro-inflammatory cytokine, interleukin (IL)-8 by target cells compared with the intact LPS. The phosphatase activity of wheat phytase towards LPS was investigated in the presence or absence of inhibitors such as L-phenylalanine and L-homoarginine. In vitro toxicity of LPS hydrolyzed with wheat phytase in comparison to intact LPS was assessed. Cell viability in human aortic endothelial (HAE) cells exposed to LPS treated with wheat phytase in comparison to intact LPS was measured. The release of IL-8 in human intestinal epithelial cell line, HT-29 cells applied to LPS treated with wheat phytase in comparison to intact LPS was assayed. Wheat phytase hydrolyzed LPS, resulting in a significant release of inorganic phosphate for 1 h (p < 0.05). Furthermore, the degradation of LPS by wheat phytase was nearly unaffected by the addition of L-phenylalanine, the inhibitor of tissue-specific alkaline phosphatase or L-homoarginine, the inhibitor of tissue-non-specific alkaline phosphatase. Wheat phytase effectively reduced the in vitro toxicity of LPS, resulting in a retention of 63% and 54% of its initial toxicity after 1-3 h of the enzyme reaction, respectively (p < 0.05). Intact LPS decreased the cell viability of HAE cells. However, LPS dephosphorylated by wheat phytase counteracted the inhibitory effect on cell viability. LPS treated with wheat phytase decreased IL-8 secretion from intestinal epithelial cell line, HT-29 cell to 14% (p < 0.05) when compared with intact LPS. In conclusion, wheat phytase is a potential therapeutic candidate and prophylactic agent for control of infections induced by pathogenic Gram-negative bacteria and associated LPS-mediated inflammatory diseases in animal husbandry.

Characterization of a Thermostable Lichenase from Bacillus subtilis B110 and Its Effects on β-Glucan Hydrolysis

  • Huang, Zhen;Ni, Guorong;Wang, Fei;Zhao, Xiaoyan;Chen, Yunda;Zhang, Lixia;Qu, Mingren
    • Journal of Microbiology and Biotechnology
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    • v.32 no.4
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    • pp.484-492
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    • 2022
  • Lichenase is an enzyme mainly implicated in the degradation of polysaccharides in the cell walls of grains. Emerging evidence shows that a highly efficient expression of a thermostable recombinant lichenase holds considerable promise for application in the beer-brewing and animal feed industries. Herein, we cloned a lichenase gene (CelA203) from Bacillus subtilis B110 and expressed it in E. coli. This gene contains an ORF of 729 bp, encoding a protein with 242 amino acids and a calculated molecular mass of 27.3 kDa. According to the zymogram results, purified CelA203 existed in two forms, a monomer, and a tetramer, but only the tetramer had potent enzymatic activity. CelA203 remained stable over a broad pH and temperature range and retained 40% activity at 70℃ for 1 h. The Km and Vmax of CelA203 towards barley β-glucan and lichenan were 3.98 mg/ml, 1017.17 U/mg, and 2.78 mg/ml, 198.24 U/mg, respectively. Furthermore, trisaccharide and tetrasaccharide were the main products obtained from CelA203-mediated hydrolysis of deactivated oat bran. These findings demonstrate a promising role for CelA203 in the production of oligosaccharides in animal feed and brewing industries.

Betulin, an Anti-Inflammatory Triterpenoid Compound, Regulates MUC5AC Mucin Gene Expression through NF-kB Signaling in Human Airway Epithelial Cells

  • Hossain, Rajib;Kim, Kyung-il;Jin, Fengri;Lee, Hyun Jae;Lee, Choong Jae
    • Biomolecules & Therapeutics
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    • v.30 no.6
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    • pp.540-545
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    • 2022
  • Betulin is a triterpenoid natural product contained in several medicinal plants including Betulae Cortex. These medicinal plants have been used for controlling diverse inflammatory diseases in folk medicine and betulin showed anti-inflammatory, antioxidative, and anticancer activities. In this study, we tried to examine whether betulin exerts a regulative effect on the gene expression of MUC5AC mucin under the status simulating a pulmonary inflammation, in human airway epithelial cells. Confluent NCI-H292 cells were pretreated with betulin for 30 min and then stimulated with phorbol 12-myristate 13-acetate (PMA) for 24 h or the indicated periods. The MUC5AC mucin mRNA expression and mucin glycoprotein production were measured by reverse transcription - polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA), respectively. To elucidate the action mechanism of betulin, effect of betulin on PMA-induced nuclear factor kappa B (NF-kB) signaling pathway was also investigated by western blot analysis. The results were as follows: 1) Betulin significantly suppressed the production of MUC5AC mucin glycoprotein and down-regulated MUC5AC mRNA expression induced by PMA in NCI-H292 cells. 2) Betulin inhibited NF-κB activation stimulated by PMA. Suppression of inhibitory kappa B kinase (IKK) by betulin led to the inhibition of the phosphorylation and degradation of inhibitory kappa B alpha (IκBα), and the nuclear translocation of NF-κB p65. This, in turn, led to the down-regulation of MUC5AC glycoprotein production in NCI-H292 cells. These results suggest betulin inhibits the gene expression of mucin through regulation of NF-kB signaling pathway, in human airway epithelial cells.

Chemical change of urushiol during heating process of Toxicodendron vernicifluum resin (Urushiol의 화학적 변화를 통한 건칠(乾漆)의 포제법(炮製法) 고찰)

  • Kim, Jung-Hoon;Doh, Eui jeong;Lee, Guemsan
    • The Korea Journal of Herbology
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    • v.35 no.2
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    • pp.1-6
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    • 2020
  • Objectives : Heating process is the traditional processing method that has been applied to reduce the toxicity of dried resin of Toxicodendron vernicifluum (Anacardiacea) used as Geon-chil (乾漆, Lacca Rhois Exsiccata or Toxicodendri Resina). Urushiol, which is found in the plants of Toxicodendron genus, is a toxic compound that is absorbed into the skin and induces allergic dermatitis by being contacted. Hence, the reduction of urushiol level by heating processing of Geon-chil is crucial method for its medicinal application. Methods : Due to lack of Geon-chil processing-related articles, the articles researching the processing of lacquer (漆), as a coating material, were collected and analyzed to investigate the chemical change of urushiol during heating process. Results : The results demonstrate that the resin which was collected from the sap of T. vernicifluum tree was dried under warm and humid conditions repeatedly. During primary drying process, the laccase, a copper-containing enzyme in the resin, participated in the formation of urushiol polymers and thereafter urushiol-related toxicity could be reduced by making a lacquer harder and more stable. Moreover, heating a lacquer over 200℃ could cause thermo-degradation of urushiol polymers, and vaporized thermally degraded urushiol monomers and their by-products, which were determined using pyrolysis/GC-MS. Conclusions : These results support that heating process being performed over 200 ℃, such as stir-frying (炒) or calcination (煅), reduces the urushiol content in Geon-chil and hence, its medicinal use can be more stable without urushiol-related allergic reactions.