Bioactive lipids in food and pharmaceutical products

食品和药品中的生物活性脂质

• 批准号: 8056822
• 负责人: ERIC J HAAS-STAPLETON
• 金额: $ 10.73万
• 依托单位: CALIFORNIA STATE UNIVERSITY LONG BEACH
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-06-01 至 2014-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8056822
• 关键词: Alcoholic Beverages; Anabolism; Arachidonic Acids; Award; Beer; Berlin; Biochemical; Biochemical Pathway; Biological; Biomedical Research; California; Candida albicans; Cell physiology; Cells; Collaborations; Cultured Cells; Development; Diagnostic; Dinoprostone; Disease; Economics; Engineering; Enzymes; Faculty; Fatty Acids; Food; Fostering; Funding; Gastrointestinal Diseases; Gastrointestinal tract structure; Gene Proteins; Generations; Genetic; Genetic Transcription; Goals; Grant; Health; High Pressure Liquid Chromatography; Hispanics; Homologous Gene; Human; Inflammation; Inflammatory; Institution; Intervention; Investigation; Life; Lipids; Liquid Chromatography; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Mentors; Methodology; Microbe; Modification; Monoclonal Antibodies; Pathway interactions; Pharmacologic Substance; Phase; Postdoctoral Fellow; Production; Property; Prostaglandin-Endoperoxide Synthase; Proteins; Publishing; Recombinant Proteins; Relative Risks; Research; Research Personnel; Resources; Reverse Transcription; Risk; Role; Saccharomyces cerevisiae; San Francisco; Signaling Molecule; Specificity; Spectrometry, Mass, Electrospray Ionization; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Staging; Therapeutic; Training; Universities; Vaccines; Work; Yeasts; career; enzyme substrate; expectation; experience; fungus; gel electrophoresis; genome sequencing; improved; innovation; interest; liquid chromatography mass spectrometry; manufacturing process; member; novel; oxidation; pathogen; protein expression; public health relevance; response; therapeutic vaccine

DESCRIPTION (provided by applicant): Prostaglandin E2 (PGE2) is a bioactive lipid signaling molecule implicated in a range of inflammatory diseases and cancers. In most human cells, PGE2 is biosynthesized via sequential oxidation of arachidonic acid (AA) by cyclooxygenases (COX) and prostaglandin E2 synthase (PTGES). At least two fungi that are pathogenic to humans also produce PGE2, which is thought to potentiate their invasive properties in humans. Our preliminary studies show that Saccharomyces cerevisiae (brewers yeast), a saprophytic fungus, also produces PGE2. PGE2 generated by S. cerevisiae is of concern because this fungus is used to manufacture a wide range of products including alcoholic beverages and recombinant proteins for therapeutics and vaccines. We investigated PGE2 levels in alcoholic beverages using monoclonal antibodies having high specificity for PGE2 and detected PGE2 in beer and sake. Moreover, in both products, we detected live yeast that biosynthesize PGE2 when provided AA as a substrate. However, the sequenced genomes of fungi, including S. cerevisiae, lack genes homologous to mammalian COX and PTGES, suggesting a novel biochemical pathway for PGE2 biosynthesis by fungi. Because PGE2 and/or PGE2 synthesizing yeast that are consumed by humans may stimulate or promote gastrointestinal (GI) disease, we propose to characterize PGE2 levels in products manufactured using S. cerevisiae with liquid chromatography-mass spectrometry. To determine the potential consequences of PGE2 on cells of the GI tract, we aim to examine the biological responses of cultured cells derived from the GI tract to the quantities of PGE2 found in products manufactured using S. cerevisiae with quantitative reverse transcription PCR. The fungal PGE2-synthesis pathway merits investigation because of its biochemical uniqueness and the potential of fungal PGE2 modulating disease in humans. Thus, we propose to define the fatty acid (FA) substrates and enzymes responsible for PGE2 biosynthesis in S. cerevisiae using genetic, biochemical and MALDI-TOF/TOF mass spectrometry methodologies. We anticipate this research has the potential to improve human health by providing a strategy to limit the introduction of FA substrates for PGE2 into manufacturing processes and point to a means for genetically engineering fungi of economic import which lack key PGE2-synthesis enzymes. PUBLIC HEALTH RELEVANCE: Prostaglandin E2 (PGE2) contained in products manufactured using S. cerevisiae (brewers yeast), such as alcoholic beverages, may elevate the risk for inflammation or cancer in the GI tract. Results obtained from the proposed studies will inform us of the relative risk to human health posed by PGE2 present in products. Understanding the substrates and biosynthetic pathways for PGE2 produced by S. cerevisiae will suggest modifications to manufacturing processes that may limit the quantities of bioactive lipids present in the final products.

描述（由申请人提供）：前列腺素E2（PGE2）是一种与一系列炎症性疾病和癌症有关的生物活性脂质信号分子。在大多数人类细胞中，PGE2是通过环氧酶（COX）和前列腺素蛋白E2合酶（PTGES）的顺序氧化通过蛛网膜酸（AA）进行生物合成的。至少有两种对人类致病的真菌也会产生PGE2，这被认为增强了它们在人类中的侵入性。我们的初步研究表明，酿酒酵母（Brewers酵母）是一种腐生真菌，也会产生PGE2。酿酒酵母产生的PGE2令人担忧，因为这种真菌用于生产各种产品，包括用于治疗和疫苗的含酒精饮料和重组蛋白。我们使用对PGE2高特异性的单克隆抗体研究了酒精饮料中的PGE2水平，并在啤酒和清酒中检测到了PGE2。此外，在这两种产品中，我们都检测到了活酵母，当提供AA作为底物时，生物合成PGE2。然而，包括酿酒酵母在内的真菌的测序基因组缺乏与哺乳动物Cox和Ptges同源的基因，这表明真菌是PGE2生物合成的新型生化途径。由于人类消耗的PGE2和/或PGE2合成的酵母可能会刺激或促进胃肠道（GI）疾病，因此我们建议在使用酿酒酵母和液相色谱质量谱法生产的产品中表征PGE2水平。为了确定PGE2对胃肠道细胞的潜在后果，我们的目的是检查使用胃肠道衍生出的培养细胞的生物学反应，该培养细胞对使用酿酒酵母制造的产品中的PGE2数量，并具有定量的逆转录转录PCR。真菌PGE2合成途径值得研究，因为它具有生化独特性和真菌PGE2调节人类疾病的潜力。因此，我们建议使用遗传，生化和MALDI-TOF/TOF/TOF质谱法方法来定义负责PGE2生物合成的脂肪酸（FA）底物和酶。我们预计，这项研究有可能通过提供一项策略来限制PGE2将FA底物引入制造过程中，并指向缺乏关键PGE2合成酶的基因工程真菌的一种手段。 公共卫生相关性：使用S. cerevisiae（Brewers酵母）制造的产品中包含的前列腺素E2（PGE2），例如酒精饮料，可能会增加胃肠道疾病或癌症的风险。从拟议的研究中获得的结果将告知我们产品中PGE2构成的人类健康的相对风险。了解酿酒酵母生产的PGE2的底物和生物合成途径将提出对制造过程的修改，以限制最终产品中存在的生物活性脂质的数量。