Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/98027
標題: 以基因體與蛋白質體學評估植生型乳酸菌株Lactobacillus brevis YW108分解酒精/乙醛能力及降乙醛產物開發
Alcohol/acetaldehyde metabolism assessment of probiotic Lactobacillus brevis YW108 by Genomic and Proteomics analysis and development of acetaldehyde reduction product
作者: 邱渝凱
Yu-Kai Chiu
關鍵字: 酒精
乙醛脫氫酶
酒精脫氫酶
alcohol
aldehyde dehydrogenase
alcohol dehydrogenase
Lactobacillus brevis YW108
Genomic
Proteomics
Phenotype MicroArrays
引用: 顏銘漢、林可寰、薛文傑:容易忽略的併發症--酒精戒斷症候群,台灣醫界2011;54:578-584。 葉娟美、廖祐毅,民105,Lactobacillus brevis YW108生化特性分析技術報告 葉娟美、蔡一瑩,民107,植生型乳酸菌株Lactobacillus brevis YW108代謝酒精之特性,對口服酒精代謝之影響以及重組人類乙醛脫氫酶於酵母菌株Kluyveromyces lactis表現之研究 Birley, A. J., M. R. James, P. A. Dickson, et al. 2007 Association of the Gastric Alcohol Dehydrogenase Gene ADH7 with Variation in Alcohol Metabolism. Human Molecular Genetics 17(2): 179–189. Cederbaum, Arthur I. 2012 Alcohol Metabolism. Clinics in Liver Disease 16(4): 667–685. Dasgupta, Amitava 2015 Chapter 2 - Genetic Aspects of Alcohol Metabolism and Drinking Behavior. In Alcohol and Its Biomarkers Pp. 37–63. San Diego: Elsevier. https://www.sciencedirect.com/science/article/pii/B978012800339800002X, accessed June 26, 2018. Edenberg, Howard J. 2007 The Genetics of Alcohol Metabolism: Role of Alcohol Dehydrogenase and Aldehyde Dehydrogenase Variants. Alcohol Research & Health 30(1): 5–13. Gross, Eric R., Vanessa O. Zambelli, Bryce A. Small, et al. 2015 A Personalized Medicine Approach for Asian Americans with the Aldehyde Dehydrogenase 2*2 Variant. Annual Review of Pharmacology and Toxicology 55(1): 107–127. Hasi, T., L. Hao, L. Yang, and X. L. Su 2011 Acetaldehyde Dehydrogenase 2 SNP Rs671 and Susceptibility to Essential Hypertension in Mongolians: A Case Control Study. Genetics and Molecular Research: GMR 10(1): 537–543. Heit, Claire, Hongbin Dong, Ying Chen, et al. 2013 The Role of CYP2E1 in Alcohol Metabolism and Sensitivity in the Central Nervous System. In Cytochrome P450 2E1: Its Role in Disease and Drug Metabolism. Aparajita Dey, ed. Pp. 235–247. Dordrecht: Springer Netherlands. http://link.springer.com/10.1007/978-94-007-5881-0_8, accessed June 2, 2018. Kvint, Kristian, Laurence Nachin, Alfredo Diez, and Thomas Nyström 2003 The Bacterial Universal Stress Protein: Function and Regulation. Current Opinion in Microbiology 6(2): 140–145. Kwak, Suryang, Yong-Cheol Park, and Jin-Ho Seo 2013 Biosynthesis of 3-Hydroxypropionic Acid from Glycerol in Recombinant Escherichia Coli Expressing Lactobacillus Brevis DhaB and DhaR Gene Clusters and E. Coli K-12 AldH. Bioresource Technology 135: 432–439. Mackie, Amanda M., Karl A. Hassan, Ian T. Paulsen, and Sasha G. Tetu 2014 Biolog Phenotype Microarrays for Phenotypic Characterization of Microbial Cells. Methods in Molecular Biology (Clifton, N.J.) 1096: 123–130. Ramírez-Nuñez, Jennifer, Ruth Romero-Medrano, Guadalupe V. Nevárez-Moorillón, and Néstor Gutiérrez-Méndez 2011 Effect of PH and Salt Gradient on the Autolysis of Lactococcus Lactis Strains. Brazilian Journal of Microbiology 42(4): 1495–1499. Stiles, Michael E., and Wilhelm H. Holzapfel 1997 Lactic Acid Bacteria of Foods and Their Current Taxonomy. International Journal of Food Microbiology 36(1): 1–29. Tilloy, Valentin, Axelle Cadière, Maryam Ehsani, and Sylvie Dequin 2015 Reducing Alcohol Levels in Wines through Rational and Evolutionary Engineering of Saccharomyces Cerevisiae. International Journal of Food Microbiology 213: 49–58. Voelkerding, K. V., S. A. Dames, and J. D. Durtschi 2009 Next-Generation Sequencing: From Basic Research to Diagnostics. Clinical Chemistry 55(4): 641–658. Zheng, Fanfan, Hao Yan, Bing Liu, et al. 2016 ALDH2 Glu504Lys Confers Susceptibility to Schizophrenia and Impacts Hippocampal-Prefrontal Functional Connectivity. Cerebral Cortex: bhw056.
摘要: 酒,自古以來一直陪伴著人類的文明前進,酒在華人的文化中佔有重要的地位,在應酬上不可或缺的就是酒,而相關研究指出,長期過度攝入酒精,容易對個人健康、家庭甚至危害他人安全。亞洲人又因基因缺陷,相當一部分的人體內代謝乙醛 ( 酒精代謝之中間物 ) 的酵素活性較差,也就是乙醛去氫酶 ( Acetaldehyde dehydrogenase,ALDH ) 的基因型突變,導致酵素活性大幅降低,乙醛更容易在體內堆積,造成一系列身體損傷。 實驗室先前由自然醱酵酸白菜醱酵汁中篩選出能夠有效代謝酒精的菌株YW108,但在進一步的人體實驗後,發現YW108確實能幫助人體代謝酒精,但代謝乙醛的速率並沒有如預期般的好,YW108在人體中反而會提高乙醛產生的速率,加速堆積,更快進入酒醉狀態。因此本實驗第一個目的就是利用酵素法分析YW108酒精代謝活性部位,再利用基因質體與蛋白質體學分析找出可能提升乙醛代謝能力的酵素,添加到YW108產品中,提升代謝乙醛能力。 酵素法首先發現乙醛誘導後的YW108胞內可溶蛋白部分具有能將乙醛代謝成乙酸的活性,後用蛋白質體學分析發現到經乙醛誘導後的YW108確實會表現多種酒精代謝相關酵素,其中兩種具有代謝乙醛能力,我們將這兩種酵素定義為目標蛋白質,未來則可利用本實驗建立的基因資料庫進行進一步的純化表現實驗,更加針對性的提升YW108產品代謝乙醛的能力。 本實驗另一個目的則是利用Phenotype MicroArrays ( PM ) 進行YW108生長源分析,目的是找出YW108最適培養條件。過去實驗室培養YW108都以MRS當作主要培養基進行培養,利用PM後得到大量YW108對各式營養源的嗜好性,這些數據在未來YW108進入工業規模生產可以用來改良培養基以提升YW108的產量。
Wine has been accompanying the progress of human civilization since ancient times. In the east, wine plays an important role in the Chinese culture, especially at sociality. According to the report, long-term excessive intake of alcohol is likely to endanger individual health and even harm others. Moreover, about1/4 of Asia people suffer from lower activity of acetaldehyde ( Intermediate of alcohol metabolism ) metabolism due to the gene mutation of acetaldehyde dehydrogenase ( ALDH ). This genetic mutation is the main reason of the significant decrease in ALDH activity and that causes the accumulate of acetaldehyde in the body, and therefore causing a series of physical damage. Previously, our lab screened a probiotic strain from fermented sour cabbage liquid. The strain was characterized and designated as Lactobacillus brevis YW108 which have ability to metabolize alcohol. However, in human trial YW108 does help human body to metabolize alcohol, but the rate of acetaldehyde metabolism is not as good as expected. Thus for people who suffering from ALDH mutation, L. brevis YW108 accelerate accumulation of acetaldehyde, and get into a drunken state more quickly. In the first part of this study, we analyze alcohol and acetaldehyde metabolic active fraction of YW108 by enzyme method. Then, genomic and proteomic analysis were used to identify enzymes that may enhance the metabolism of acetaldehyde We found that the intracellular soluble protein fraction of YW108 induced by acetaldehyde has the activity of metabolizing acetaldehyde by enzymeatic analysis. Using proteomic analysis, it was found that YW108 induced by acetaldehyde does exhibit a variety of enzymes related to alcohol metabolism, two of which have the ability to metabolize acetaldehyde. We define these two enzymes as target proteins which can seen as candidate for acetaldehyde reduction product development. In another part of this study, we used Phenotype MicroArrays ( PM ) for YW108 growth source analysis to find the optimal culture conditions. In the future, we can use these data to improve the medium to increase the yield of YW108 when it enters industrial scale production.
URI: http://hdl.handle.net/11455/98027
文章公開時間: 2021-08-27
Appears in Collections:食品暨應用生物科技學系

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