本文主要研究内容
作者刘亚楠（2019）在《蛋白乙酰化对酿酒酵母碳代谢的影响初探》一文中研究指出：糖(碳源)代谢过程是生物体最重要的代谢过程,其往往受到严格的信号调控。当其代谢过程出现紊乱时,往往导致细胞不同程度的病变、死亡。酿酒酵母是研究较为深入的真核模式生物,具有食品级安全、代谢旺盛等特点,非常适合作为多种发酵产品生产的细胞工厂,深入研究其代谢调控机制将有助于开发新的构建策略,促进其在代谢工程、合成生物学领域的应用。酿酒酵母糖代谢和大部分真核生物一样,主要包括糖酵解途径、磷酸戊糖途径、TCA循环、乙醛酸循环和糖异生途径等;另一方面,其代谢又具有典型的特点。当生长环境中存在丰富发酵性碳源(最典型的:葡萄糖)的条件下,无论供氧充足与否,酿酒酵母细胞都处于强酵解状态,只有少量丙酮酸进入TCA循环进行呼吸代谢,而大部分生成乙醇。当葡萄糖耗尽,酵母通过级联的信号途径做出响应,从转录、翻译、翻译后蛋白质修饰等各个层面调节各大代谢途径中基因的表达和蛋白活性,从而精确调控代谢网络转为呼吸代谢模式,利用发酵葡萄糖时积累的非发酵性碳源——乙醇进行二次生长。这种碳源切换引起的蛋白表达修饰和生理代谢变化过程涉及生命活动最基本的对环境营养的响应和对自身代谢的调控,而被科学家们长期关注。目前对这些过程涉及的信号系统以及调节机制主要包括对细胞整体代谢酶的转录调控和蛋白磷酸化修饰以及相关的调控通路和反馈回路,仍然处于有限认知的水平。赖氨酸乙酰化是细胞内一种重要的翻译后修饰过程。多年来,相关研究主要集中在组蛋白乙酰化修饰对基因转录水平的调节、DNA的复制和修复等,关注的是主要是其表观遗传学意义,也有少部分工作涉及到了代谢等生理特征等表型现象。另一方面,最近的研究发现,除组蛋白外,大量代谢酶蛋白也存在乙酰化位点。基于此,本论文从代谢酶在不同碳源条件下的乙酰化差异和扰动乙酰化和脱乙酰化过程对碳代谢的影响两个角度,初探在调控水平乙酰化对酿酒酵母碳代谢的影响。本论文第一部分工作首先利用高分辨质谱解析发酵性碳源葡萄糖以及非发酵性碳源乙醇这两种碳源条件下酿酒酵母乙酰化组的差异。结果显示,在乙醇条件下,糖酵解途径、戊糖磷酸途径、核糖体蛋白的乙酰化水平多呈现下调趋势:三羧酸循环中蛋白乙酰化水平呈现上调趋势。之后,选择在不同碳源条件下存在乙酰化差异的代谢途径蛋白和糖信号途径蛋白,利用点突变技术,将乙酰化位点处赖氨酸突变为精氨酸,使该乙酰化位点成为组成型非乙酰化状态。继而,通过生化水平的酶学测定,研究具体的乙酰化修饰位点对蛋白活性的影响。结果显示,分别与对照相比,丙酮酸激酶(编码基因PYK1)、苹果酸脱氢酶(编码基因MDH1)、葡萄糖-6-磷酸脱氢酶(编码基因ZWF1)乙酰化位点突变菌株比酶活分别提高了25.6%、41.4%、27.3%;而果糖1,6-二磷酸醛缩酶(编码基因FBA1)乙酰化位点突变菌株比酶活降低了 19.9%。上述代谢酶单个酶活性水平的变化不影响菌株的发酵特征。但是,乙醇脱氢酶(编码基因ADH1)乙酰化位点突变后,菌株发酵葡萄糖时菌体得率明显降低,而乙醇得率增加,乙酸明显积累。这些结果表明,酿酒酵母中,乙酰化会影响代谢酶活性,某些酶如Adh1突变子能够显著影响代谢。多个酶蛋白乙酰化差异带来的整体调控效应还有待进一步论证。课题第二部分通过敲除乙酰化酶或去乙酰化酶的催化亚基或重要组成亚基,人为扰动乙酰化过程并研究了这些扰动对菌株代谢特征的影响。结果显示,敲除GCN5、ADA2对菌体得率没有影响,对葡萄糖利用没有促进作用,乙醇得率略有增加;敲除NAT3降低了菌体得率,对葡萄糖利用没有促进作用,乙醇得率略有增加;敲除HOS2、HST2、SET3、RCO1、SDS3、RPD3没有对葡萄糖的利用和乙醇产生没有明显影响。敲除ARD1、SAS3加快了发酵代谢阶段葡萄糖的利用,降低菌体得率,增加乙醇得率;敲除HAT1(目前酵母中唯一已知的细胞质乙酰基转移酶)能够加快呼吸代谢阶段乙醇的利用。这些结果表明,乙酰化参与酿酒酵母的细胞代谢调控,但具体机制还有待进一步研究。现有的代谢组、转录组、蛋白组、磷酸化组等研究对酿酒酵母碳代谢的揭示已经相对比较深入,但有关代谢的调节机制仍有很多未知。近年来,有观点认为乙酰化调节细胞生理代谢具有从细菌到人类的保守性。本论文的研究是有关蛋白乙酰化调节代谢研究的有益补充,具有揭示生物界代谢机制和支持进化观点的广泛意义。
Abstract
tang (tan yuan )dai xie guo cheng shi sheng wu ti zui chong yao de dai xie guo cheng ,ji wang wang shou dao yan ge de xin hao diao kong 。dang ji dai xie guo cheng chu xian wen luan shi ,wang wang dao zhi xi bao bu tong cheng du de bing bian 、si wang 。niang jiu jiao mu shi yan jiu jiao wei shen ru de zhen he mo shi sheng wu ,ju you shi pin ji an quan 、dai xie wang cheng deng te dian ,fei chang kuo ge zuo wei duo chong fa jiao chan pin sheng chan de xi bao gong an ,shen ru yan jiu ji dai xie diao kong ji zhi jiang you zhu yu kai fa xin de gou jian ce lve ,cu jin ji zai dai xie gong cheng 、ge cheng sheng wu xue ling yu de ying yong 。niang jiu jiao mu tang dai xie he da bu fen zhen he sheng wu yi yang ,zhu yao bao gua tang jiao jie tu jing 、lin suan wu tang tu jing 、TCAxun huan 、yi quan suan xun huan he tang yi sheng tu jing deng ;ling yi fang mian ,ji dai xie you ju you dian xing de te dian 。dang sheng chang huan jing zhong cun zai feng fu fa jiao xing tan yuan (zui dian xing de :pu tao tang )de tiao jian xia ,mo lun gong yang chong zu yu fou ,niang jiu jiao mu xi bao dou chu yu jiang jiao jie zhuang tai ,zhi you shao liang bing tong suan jin ru TCAxun huan jin hang hu xi dai xie ,er da bu fen sheng cheng yi chun 。dang pu tao tang hao jin ,jiao mu tong guo ji lian de xin hao tu jing zuo chu xiang ying ,cong zhuai lu 、fan yi 、fan yi hou dan bai zhi xiu shi deng ge ge ceng mian diao jie ge da dai xie tu jing zhong ji yin de biao da he dan bai huo xing ,cong er jing que diao kong dai xie wang lao zhuai wei hu xi dai xie mo shi ,li yong fa jiao pu tao tang shi ji lei de fei fa jiao xing tan yuan ——yi chun jin hang er ci sheng chang 。zhe chong tan yuan qie huan yin qi de dan bai biao da xiu shi he sheng li dai xie bian hua guo cheng she ji sheng ming huo dong zui ji ben de dui huan jing ying yang de xiang ying he dui zi shen dai xie de diao kong ,er bei ke xue jia men chang ji guan zhu 。mu qian dui zhe xie guo cheng she ji de xin hao ji tong yi ji diao jie ji zhi zhu yao bao gua dui xi bao zheng ti dai xie mei de zhuai lu diao kong he dan bai lin suan hua xiu shi yi ji xiang guan de diao kong tong lu he fan kui hui lu ,reng ran chu yu you xian ren zhi de shui ping 。lai an suan yi xian hua shi xi bao nei yi chong chong yao de fan yi hou xiu shi guo cheng 。duo nian lai ,xiang guan yan jiu zhu yao ji zhong zai zu dan bai yi xian hua xiu shi dui ji yin zhuai lu shui ping de diao jie 、DNAde fu zhi he xiu fu deng ,guan zhu de shi zhu yao shi ji biao guan wei chuan xue yi yi ,ye you shao bu fen gong zuo she ji dao le dai xie deng sheng li te zheng deng biao xing xian xiang 。ling yi fang mian ,zui jin de yan jiu fa xian ,chu zu dan bai wai ,da liang dai xie mei dan bai ye cun zai yi xian hua wei dian 。ji yu ci ,ben lun wen cong dai xie mei zai bu tong tan yuan tiao jian xia de yi xian hua cha yi he rao dong yi xian hua he tuo yi xian hua guo cheng dui tan dai xie de ying xiang liang ge jiao du ,chu tan zai diao kong shui ping yi xian hua dui niang jiu jiao mu tan dai xie de ying xiang 。ben lun wen di yi bu fen gong zuo shou xian li yong gao fen bian zhi pu jie xi fa jiao xing tan yuan pu tao tang yi ji fei fa jiao xing tan yuan yi chun zhe liang chong tan yuan tiao jian xia niang jiu jiao mu yi xian hua zu de cha yi 。jie guo xian shi ,zai yi chun tiao jian xia ,tang jiao jie tu jing 、wu tang lin suan tu jing 、he tang ti dan bai de yi xian hua shui ping duo cheng xian xia diao qu shi :san suo suan xun huan zhong dan bai yi xian hua shui ping cheng xian shang diao qu shi 。zhi hou ,shua ze zai bu tong tan yuan tiao jian xia cun zai yi xian hua cha yi de dai xie tu jing dan bai he tang xin hao tu jing dan bai ,li yong dian tu bian ji shu ,jiang yi xian hua wei dian chu lai an suan tu bian wei jing an suan ,shi gai yi xian hua wei dian cheng wei zu cheng xing fei yi xian hua zhuang tai 。ji er ,tong guo sheng hua shui ping de mei xue ce ding ,yan jiu ju ti de yi xian hua xiu shi wei dian dui dan bai huo xing de ying xiang 。jie guo xian shi ,fen bie yu dui zhao xiang bi ,bing tong suan ji mei (bian ma ji yin PYK1)、ping guo suan tuo qing mei (bian ma ji yin MDH1)、pu tao tang -6-lin suan tuo qing mei (bian ma ji yin ZWF1)yi xian hua wei dian tu bian jun zhu bi mei huo fen bie di gao le 25.6%、41.4%、27.3%;er guo tang 1,6-er lin suan quan su mei (bian ma ji yin FBA1)yi xian hua wei dian tu bian jun zhu bi mei huo jiang di le 19.9%。shang shu dai xie mei chan ge mei huo xing shui ping de bian hua bu ying xiang jun zhu de fa jiao te zheng 。dan shi ,yi chun tuo qing mei (bian ma ji yin ADH1)yi xian hua wei dian tu bian hou ,jun zhu fa jiao pu tao tang shi jun ti de lv ming xian jiang di ,er yi chun de lv zeng jia ,yi suan ming xian ji lei 。zhe xie jie guo biao ming ,niang jiu jiao mu zhong ,yi xian hua hui ying xiang dai xie mei huo xing ,mou xie mei ru Adh1tu bian zi neng gou xian zhe ying xiang dai xie 。duo ge mei dan bai yi xian hua cha yi dai lai de zheng ti diao kong xiao ying hai you dai jin yi bu lun zheng 。ke ti di er bu fen tong guo qiao chu yi xian hua mei huo qu yi xian hua mei de cui hua ya ji huo chong yao zu cheng ya ji ,ren wei rao dong yi xian hua guo cheng bing yan jiu le zhe xie rao dong dui jun zhu dai xie te zheng de ying xiang 。jie guo xian shi ,qiao chu GCN5、ADA2dui jun ti de lv mei you ying xiang ,dui pu tao tang li yong mei you cu jin zuo yong ,yi chun de lv lve you zeng jia ;qiao chu NAT3jiang di le jun ti de lv ,dui pu tao tang li yong mei you cu jin zuo yong ,yi chun de lv lve you zeng jia ;qiao chu HOS2、HST2、SET3、RCO1、SDS3、RPD3mei you dui pu tao tang de li yong he yi chun chan sheng mei you ming xian ying xiang 。qiao chu ARD1、SAS3jia kuai le fa jiao dai xie jie duan pu tao tang de li yong ,jiang di jun ti de lv ,zeng jia yi chun de lv ;qiao chu HAT1(mu qian jiao mu zhong wei yi yi zhi de xi bao zhi yi xian ji zhuai yi mei )neng gou jia kuai hu xi dai xie jie duan yi chun de li yong 。zhe xie jie guo biao ming ,yi xian hua can yu niang jiu jiao mu de xi bao dai xie diao kong ,dan ju ti ji zhi hai you dai jin yi bu yan jiu 。xian you de dai xie zu 、zhuai lu zu 、dan bai zu 、lin suan hua zu deng yan jiu dui niang jiu jiao mu tan dai xie de jie shi yi jing xiang dui bi jiao shen ru ,dan you guan dai xie de diao jie ji zhi reng you hen duo wei zhi 。jin nian lai ,you guan dian ren wei yi xian hua diao jie xi bao sheng li dai xie ju you cong xi jun dao ren lei de bao shou xing 。ben lun wen de yan jiu shi you guan dan bai yi xian hua diao jie dai xie yan jiu de you yi bu chong ,ju you jie shi sheng wu jie dai xie ji zhi he zhi chi jin hua guan dian de an fan yi yi 。
论文参考文献
论文详细介绍
论文作者分别是来自山东大学的刘亚楠，发表于刊物山东大学2019-07-16论文，是一篇关于酿酒酵母论文,碳代谢论文,蛋白乙酰化修饰论文,发酵代谢论文,呼吸代谢论文,山东大学2019-07-16论文的文章。本文可供学术参考使用，各位学者可以免费参考阅读下载，文章观点不代表本站观点，资料来自山东大学2019-07-16论文网站，若本站收录的文献无意侵犯了您的著作版权，请联系我们删除。
标签：酿酒酵母论文; 碳代谢论文; 蛋白乙酰化修饰论文; 发酵代谢论文; 呼吸代谢论文; 山东大学2019-07-16论文;