| A Procedure for Precise Determination of Glutathione Produced by Saccharomyces cerevisiae
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Author:
Date:
2018-06-20
[Abstract] In bioproduction, yields of products must be calculated precisely for accurate evaluation of various fermentation conditions. To evaluate productivity of microorganisms, product amounts per unit of medium volume (e.g., mg-product/L-broth), and/or product amounts per unit of a microorganism amount (e.g., mg-product/mg-dry cell weight) are often used. Nonetheless, detailed procedures for calculation of these production yields are often omitted in research articles, whereas methods for product quantification are described well. Here, we describe a detailed calculation procedure from our previous studies on glutathione production by Saccharomyces cerevisiae. This procedure can be applied to various other products and microorganisms, and therefore, may prove to be ...
[摘要] 在生物生产中，必须精确计算产品的产量，以准确评估各种发酵条件。 为了评估微生物的生产力，每单位培养基体积的产物量（例如，mg-产物/ L-肉汤）和/或每单位微生物量的产物量（例如， ，毫克产品/毫克干细胞重量）经常使用。 尽管如此，在研究文章中常常忽略用于计算这些产量的详细程序，而产品量化的方法则被很好地描述。 在这里，我们描述了我们以前关于酿酒酵母产生谷胱甘肽的研究的详细计算过程。 该程序可以应用于各种其他产品和微生物，因此可能证明可用于各种其他生物生产研究。
【背景】谷胱甘肽是所有生物体中含量最高的含巯基三肽，并且作为在细胞中具有不同作用的生物活性物质起作用，例如作为氧化还原和解毒剂。因此，谷胱甘肽如今被广泛用于医疗，食品和化妆品行业，并且近年来需求增加。谷胱甘肽在工业上主要通过使用原始含有高浓度谷胱甘肽的酿酒酵母进行发酵生产，并且已经作为安全的食品生产微生物。对各种微生物中的微生物谷胱甘肽产生的研究在未来将变得更加重要。为了评估各种微生物发酵产生谷胱甘肽的效率，我们在这里描述了我们详细的样品制备程序，高效色谱（HPLC）定量还原和氧化谷胱甘肽的方法，以及两种产量的计算方法（Hara ） 2012; Hara等人，2015; Kiriyama等人，2013; Kobayashi等人 2017年）。
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| In vitro Treatment of Mouse and Human Cells with Endogenous Ligands for Activation of the Aryl Hydrocarbon Receptor
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Author:
Date:
2017-01-05
[Abstract] Activation of the aryl hydrocarbon receptor (AHR) by endogenous ligands has been implicated in a variety of physiological processes such as cell cycle regulation, cell differentiation and immune responses. It is reported that tryptophan metabolites, such as kynurenine (Kyn) and 6-formylindolo(3,2-b)carbazole (FICZ), are endogenous ligands for AHR (Stockinger et al., 2014). This protocol is designed for treatment with Kyn or FICZ in mouse embryonic fibroblasts (MEFs) or primary peripheral monocytes.
[摘要] 通过内源性配体活化芳基烃受体（AHR）已涉及多种生理过程，如细胞周期调控，细胞分化和免疫应答。据报道，色氨酸代谢物，如犬尿胆碱（Kyn）和6-甲基吲哚（3,2-b）咔唑（FICZ）是AHR的内源性配体（Stockinger等人，2014）。该方案设计用于在小鼠胚胎成纤维细胞（MEF）或初级周边单核细胞中用Kyn或FICZ进行治疗。
背景 色氨酸代谢物如Kyn和FICZ是生理条件下AHR的内源性配体。 Kyn由色氨酸-2,3-双加氧酶（TDO）和/或吲哚胺-2,3-双加氧酶1和2（IDO1 / 2）产生，并有助于抑制抗肿瘤反应和恶性进展（Stockinger等人，2014）。 ...
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| HBV Infection in Human Hepatocytes and Quantification of Encapsidated HBV DNA
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Author:
Date:
2016-01-20
[Abstract] Human hepatic cancer cell lines such as HepG2, Huh7, and HLE cannot get infected with Hepatitis B virus (HBV) due to lack of an HBV receptor(s). Transfection with HBV genome has so far been referred as a tool to mimic HBV infection. However, since sodium taurocholate cotransporting polypeptide (NTCP) was identified as a functional receptor for HBV (Yan et al., 2012), hepatocyte cell lines that were stably transfected with a plasmid for NTCP expression have been used for HBV infection. This protocol is designed for infection with HBV in human hepatocyte cell line HepG2 expressing NTCP (HepG2-hNTCP-C4 cells; Iwamoto et al., 2014) or primary human hepatocytes (PHHs). In this section, we also describe one of the methods for the assessment of HBV infection: Quantification of ...
[摘要] 人肝癌细胞系如HepG2，Huh7和HLE由于缺乏HBV受体而不能感染乙型肝炎病毒（HBV）。 HBV基因组的转染迄今为止被称为模拟HBV感染的工具。 然而，由于牛磺胆酸钠共转运多肽（NTCP）被鉴定为HBV的功能性受体（Yan等人，2012），已经使用用用于NTCP表达的质粒稳定转染的肝细胞细胞系 为HBV感染。 该方案设计用于在表达人类肝细胞细胞系HepG2的表达NTCP（HepG2-hNTCP-C4细胞; Iwamoto等人，2014）或原代人肝细胞（PHH）的HBV感染。 在本节中，我们还描述了用于评估HBV感染的方法之一：细胞内衣壳化的HBV DNA的定量。
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