CN104357415A - CemGPDH基因及其应用 - Google Patents
CemGPDH基因及其应用 Download PDFInfo
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- CN104357415A CN104357415A CN201410547380.0A CN201410547380A CN104357415A CN 104357415 A CN104357415 A CN 104357415A CN 201410547380 A CN201410547380 A CN 201410547380A CN 104357415 A CN104357415 A CN 104357415A
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Classifications
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- C—CHEMISTRY; METALLURGY
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- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/0004—Oxidoreductases (1.)
- C12N9/0006—Oxidoreductases (1.) acting on CH-OH groups as donors (1.1)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
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- C12P7/649—Biodiesel, i.e. fatty acid alkyl esters
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
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- C12Y101/05—Oxidoreductases acting on the CH-OH group of donors (1.1) with a quinone or similar compound as acceptor (1.1.5)
- C12Y101/05003—Glycerol-3-phosphate dehydrogenase (1.1.5.3)
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/10—Biofuels, e.g. bio-diesel
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
本发明提供了一种CemGPDH基因及其在提高细胞脂肪酸含量方面的用途，所述CemGPDH基因的核酸序列如SEQ ID NO:1所示。该基因来源于椭圆小球藻(Chlorella ellipsoidea)，编码甘油-3-磷酸脱氢酶。甘油-3-磷酸脱氢酶(Glycerol-3-phosphate dehydrogenase，GPDH)催化合成的甘油-3-磷酸(Glycerol-3-phosphate，G3P)是合成三酰甘油(TAG)的重要原料，该酶参与了线粒体G3P穿梭，为呼吸链提供电子，是G3P合成的关键酶，也是连接糖代谢和脂代谢的关键酶之一，对植物体内油脂合成和能量代谢有重要作用。利用本发明的基因转化酵母细胞、植物细胞和微藻细胞能显著提高细胞的总脂肪酸含量。
Description
技术领域
本发明涉及一种甘油-3-磷酸脱氢酶基因GPDH及其应用。具体地，涉及该基因序列的获得和酵母表达载体的构建，以及其显著提高酵母脂肪酸含量方面的用途。
背景技术
目前，随着传统能源的过度消耗和日益增长的能源需求，能源问题日益严峻，可替代能源的研究和开发成为全球学者关注的热点。生物柴油主要以大豆、麻疯树等油料作物、废弃的厨余油脂、动物性脂肪以及基因工程改造过的微藻作为原材料加工成液体燃料，是可以替代传统石油燃料的优质新型可再生能源。而微藻由于含油量高，易于培养，单位土地油脂产量远高于其他油料作物，被认为是最有潜力的能源资源之一。
微藻繁殖能力强，对生长环境要求低，易于培养，光合作用效率高，含油量高，但目前微藻油脂代谢途径中关键酶基因的研究还相对较少[1-2]。小球藻(Chlorella)是微藻中的一类，属于绿藻门小球藻属，是光合自养型球形单细胞绿藻，直径大约3-8微米。小球藻繁殖能力很强，含有丰富的营养元素，例如蛋白质、油脂、多不饱和脂肪酸、维生素、矿物质、食物纤维、核酸及叶绿素等，还含有多种生物活性物质，例如糖蛋白、多糖。小球藻是少数几个可用于大规模培养的微藻。
真核生物(例如小球藻)中，油脂的主要储存形式是三酰甘油(TAG)，由甘油-3-磷酸(G3P)和脂酰辅酶A为前体合成。甘油-3-磷酸脱氢酶(Glycerol-3-phosphate dehydrogenase，GPDH)催化糖酵解过程中产生的磷酸二羟丙酮(Dihydroxyacetone phosphate，DHAP)与G3P之间的相互转变，产物G3P是合成TAG的重要原料，因此，GPDH是连接糖代谢和脂代谢的关键酶。
G3P是由甘油磷酸化而来，在植物中，淀粉通常作为光合作用的终产物被储存起来，甘油的主要来源是通过将淀粉分解的葡萄糖送入糖酵解过程，将糖酵解中间产物转化为甘油。而在微藻中，有些种类的微藻在积累淀粉的同时会产生大量的甘油，作为光合作用的终产物储存起来[3]。因此，在微藻中，主要有两条不同的甘油合成途径，其一是利用光合作用直接合成甘油，另一条是通过分解储藏物质，将淀粉及蔗糖分解成为6-磷酸果糖，后转变为DHAP，再由甘油-3-磷酸脱氢酶催化，生成G3P，最后通过去磷酸化作用生成甘油[4-5]。从微藻GPDH在油脂合成代谢中的位置上来看，GPDH所催化的由DHAP转变为G3P的反应是甘油合成途径的关键步骤，GPDH参与了线粒体G3P穿梭，为呼吸链提供电子，是G3P合成的关键酶之一，而G3P是TAG和磷脂合成的重要原料，并且GPDH连接了糖代谢和脂代谢，对植物体内油脂合成和能量代谢有重要作用。
根据GPDH在细胞内位置不同，将其分为三种类型同工酶——细胞质型GPDH(ctGPDH)，叶绿体型GPDH(cpGPDH)和线粒体型GPDH(mGPDH)。每种同工酶在细胞内偏好催化DHAP与G3P之间相互转变的正反应或逆反应，辅酶类型不同(以FAD或NADH/NADPH作为辅酶)，受不同条件诱导表达(渗透胁迫、热激、油酸胁迫等)，并且在不同物种中目前已发现的GPDH同工酶数量不同[5]。
发明内容
本发明发现将来源于椭圆小球藻的CemGPDH基因转入酿酒酵母INVSC1后，可显著提高其主要脂肪酸组分的含量。在INVScI主要的四种脂肪酸组分中，棕榈酸(C16:0)和硬脂酸(C18:0)含量变化不显著，棕榈油酸(C16:1)提高了15.57％，油酸(C18:1)提高了13.53％。酵母细胞总脂肪酸的含量也提高了约10.07％。已有物种GPDH基因的研究基本都选择了酵母GPDH突变体来对其进行互补功能验证，甚少有用野生型酵母的。本发明选择野生型酵母在对椭圆小球藻GPDH基因进行功能验证的同时还发现了其对油脂合成的显著作用。在酵母中研究表明，CemGPDH基因转酵母株与对照组(转空载)相比，总脂肪酸含量提高了约10.07％，且其部分主要脂肪酸组分的含量也有明显提高，其中棕榈酸(C16:0)和硬脂酸(C18:0)含量变化不显著，棕榈油酸(C16:1)提高了15.57％，油酸(C18:1)提高了13.53％。酵母细胞总脂肪酸的含量也提高了约10.07％。单个基因使得总脂肪酸及各不同脂肪酸的含量达到这种程度的实属少见。
本发明提供了一个GPDH基因，其cDNA核苷酸序列如SEQ ID NO:1所示，这个基因来源于椭圆小球藻，该基因cDNA全长为1857bp。
本发明还提供了含本发明所述基因(CemGPDH)的酵母表达载体。
本发明在一个方面提供了包含本发明所述基因的酵母、植物或藻类，其选自酵母，藻类，拟南芥、烟草、油菜、向日葵、大豆、蓖麻、棉花、橄榄、和红花等。优选地，所述藻类选自小球藻。更具体地，所述酵母选自酿酒酵母，小球藻选自椭圆小球藻(Chlorella ellipsoidea)。
本发明提供的椭圆小球藻GPDH基因，可应用于利用基因工程方法提高酵母、植物或藻类油脂等方面。
更具体地，本发明提供以下各项：
1.CemGPDH蛋白，其氨基酸序列如SEQ ID NO:2所示。
2.编码根据1所述的CemGPDH蛋白的基因。
3.根据2所述的基因，其核酸序列如SEQ ID NO:1所示。
4.载体，其包含根据2或3所述的基因，所述载体可以通过将根据2或3所述的基因转入到pYES2.0(可购自Invitrogen公司)，pBIN19、pBI121、pBI221(可购自Clontech)，pCambia1300(可购自Cambia公司)或pGreen(可购自the John Innes Centre)中来制备。
5.宿主细胞，其包含根据1所述的CemGPDH蛋白、根据2或3所述的基因或根据4所述的载体。
6.一种制备具有高棕榈油酸和/或油酸含量，或具有高总脂肪酸含量的酵母、植物或藻类的细胞的方法，所述方法包括将根据2或3所述的基因或根据4所述的载体转入所述酵母、植物或藻类的细胞中，所述酵母优选是酿酒酵母，更优选是尿嘧啶缺陷型酿酒酵母INVSC1；所述藻类优选是小球藻，更优选是椭圆小球藻(Chlorella ellipsoidea)；所述植物优选选自拟南芥、烟草、油菜、向日葵、大豆、番茄、蓖麻、棉花、芝麻或花生。
7.根据1所述的CemGPDH蛋白、根据2或3所述的基因或根据4所述的载体在制备具有高总脂肪酸含量的酵母、植物或藻类的细胞中的用途，所述酵母优选是酿酒酵母，更优选是尿嘧啶缺陷型酿酒酵母INVSC1；所述藻类优选是小球藻，更优选是椭圆小球藻(Chlorella ellipsoidea)；所述植物优选选自拟南芥、烟草、油菜、向日葵、大豆、番茄、蓖麻、棉花、芝麻或花生，所述具有高总脂肪酸含量的酵母、植物或藻类的细胞优选是具有高棕榈油酸和/或油酸含量的酵母、植物或藻类的细胞。
8.根据1所述的CemGPDH蛋白、根据2或3所述的基因或根据4所述的载体在提高酵母、植物或藻类的细胞中棕榈油酸和/或油酸含量，或总脂肪酸含量中的用途，所述酵母优选是酿酒酵母，更优选是尿嘧啶缺陷型酿酒酵母INVSC1；所述藻类优选是小球藻，更优选是椭圆小球藻(Chlorella ellipsoidea)；所述植物优选选自拟南芥、烟草、油菜、向日葵、大豆、番茄、蓖麻、棉花、芝麻或花生。
9.利用根据6所述的方法制备的具有高棕榈油酸和/或油酸含量，或具有高总脂肪酸含量的酵母、植物或藻类的细胞。
10.根据9所述的酵母、植物或藻类的细胞在生产生物柴油中的用途。
附图说明
图1.含椭圆小球藻CemGPDH基因的酵母表达载体的构建。
图2.GC-MS法测定的酵母各脂肪酸组分含量以及总脂肪酸含量(CK为对照，pYES-GPDH为转CemGPDH基因的酵母脂肪酸测定)。
具体实施方式
下面参考实施例和附图详细描述本发明。本领域的普通技术人员可以理解的是，下述实施例是举例说明的目的，其不应以任何方式解释为对本发明的限制。本发明的保护范围由后附的所限定。
实施例1.椭圆小球藻CemGPDH基因cDNA全长的获得
取培养3-4天的对数生长期椭圆小球藻(来自于中国科学院水生生物研究所)藻液，离心收集藻体后置于液氮中充分研磨。参照艾德莱EASYspinRNA提取试剂盒说明书及Takara DNase I说明书进行总RNA提取和纯化。以带polyT的引物进行逆转录获得cDNA，并进行RT-PCR(具体操作见TOYOBO ReverTra Ace-α试剂盒)。
cDNA的合成反应条件如下：
用RNase Free H2O补至总体积为20μL。轻弹混匀并瞬时离心后按如下程序进行逆转录反应：
取适量上述逆转录产物以ATGAAGTTGTCAAGATTTAC和CTATGTCTTGACTGGGGTC为一对引物进行PCR扩增。
PCR反应条件如下：
PCR产物电泳(1％琼脂糖凝胶浓度)后切胶回收目的片段(CemGPDH，约1.9kb)，连入pEASY-Blunt载体(购自Transgen公司)，测序验证获得了CemGPDH的全长cDNA(SEQ ID NO:1)。
实施例2.含有CemGPDH基因的酵母表达载体的构建
根据CemGPDH的cds序列，设计带有KpnI和EcoR I酶切位点的引物(两条引物的序列分别为：cgccGGTACCATGAAGTTGTCAAGATTTAC和ccggGAATTCCTATGTCTTGACTGGGGTC从含有CemGPDH的pEASY-Blunt载体上，采用高保真EasyPfu DNA Polymerase(购自Transgen公司)，扩增出含KpnI和EcoR I酶切位点的片段，用KpnI和EcoR I(购自Takara公司)进行双酶切，与同样双酶切的酵母表达载体pYES2(购自Invitrogen公司)连接，测序验证，并将其命名为pYES-GPDH，其载体图见图1。
实施例3.酵母表达载体pYES-GPDH转化酿酒酵母
接种尿嘧啶缺陷型酿酒酵母INVSC1(购自Invitrogen公司)于10mLYPD培养基中，30℃震荡培养过夜。次日将菌液接种到50mL YPD培养基中，稀释到OD600＝0.4，继续培养2-4h至OD600在0.4-0.6之间，5,000rpm冷冻离心1min,用40mL1×TE(10mM Tris，pH7.5，1mM EDTA)悬浮沉淀，5,000rpm冷冻离心，用2mL1×LiAc(10mM乙酸锂，pH7.5)/0.5×TE悬浮沉淀，室温孵育10min。将100μL酵母悬浮液与1ug酵母表达载体pYES-GPDH和100ug变性鲑鱼精DNA混匀，然后加入700μL1×LiAc/40％PEG-3350/1×TE，混匀。30℃培养30min，加入88μL DMSO，混匀，42℃热激7min。10000rpm离心10s，去上清，用1mL1×TE悬浮沉淀，10,000rpm离心10s，去除上清。用50-100μL1×TE悬浮沉淀，涂布于SC-U基本培养基上，30℃培养3-4天。
3-4天后，从培养基平板上挑取菌落，参考博迈德酵母高纯度质粒小量快速提取试剂盒说明书，进行质粒提取，然后将提取的质粒作为模板进行PCR验证。最后对验证为阳性的酵母质粒和相应的菌落进行保存。
实施例4.CemGPDH在酿酒酵母中的诱导表达
将转化有pYES-GPDH的酿酒酵母INVSC1的单菌落接种于5-10mLSC-U培养基中，200rpm，30℃震荡培养24-48小时。取培养的菌液转接到50mL含1％棉子糖和2％酵母表达诱导物D-半乳糖的SC-U培养液中，使其OD600约为0.1，加入NP-40(终浓度为1％，有利于酵母细胞悬浮)，200rpm，20℃，培养72h诱导表达。将转化有pYES2空载的酵母转化子设为对照，对作为对照的酵母转化子进行同样操作。
实施例5.酵母脂肪酸的提取与检测
1.酵母脂肪酸的提取
取诱导培养好的酵母液，4000rpm离心5min，室温收集菌体；经去离子水反复悬浮、室温离心洗涤三次，50℃烘干；取100mg酵母干粉充分研磨，加入3mL7.5％KOH-CH3OH，加15-20μL的d17:0(购自sigma公司，浓度27mg/mL)，70℃水浴3-5h；加入2mL HCl酸化至其pH值达2.0；加入2mL14％BF3-CH3OH(购自Aldtich公司)溶液，70℃水浴1.5h；加入1mL0.9％NaCl溶液，4mL正己烷抽提一次，N2吹干；300μL乙酸乙酯溶解。该实验每次每个样品平行做两份，共重复三次。
2.终产物GC-MS检测分析实验
所用GC/MS仪为TurboMass(PerkinElmer公司)；GC条件：色谱柱：BPX-70，30m×0.25mm×0.25um。柱温：120℃，气化室温度230℃。取1μL终产物上样，分流比10:1。
3.GC-MS结果分析
研究表明，CemGPDH基因转酵母株与对照组(转pYES2空载)相比，棕榈酸(C16:0)和硬脂酸(C18:0)含量变化不显著，棕榈油酸(C16:1)提高了15.57％，油酸(C18:1)提高了13.53％。酵母细胞总脂肪酸的含量也提高了约10.07％。GC-MS测定的酵母各脂肪酸组分及含量测定结果见图2。
参考文献：
1.Harun,R.,Singh,M.,Forde,G.M.,and Danquah,M.K.(2010).Bioprocess engineering of microalgae to produce a variety of consumerproducts.Renew Sust Energ Rev14,1037-1047.
2.Widjaja,A.,Chien,C.C.,and Ju,Y.H.(2009).Study of increasing lipidproduction from fresh water microalgae Chlorella vulgaris.J Taiwan InstChem E40,13-20.
3.Metting,F.B.(1996).Biodiversity and application of microalgae.J IndMicrobiol Biot17,477-489.
4.Chen,H.,Lao,Y.M.,and Jiang,J.G.(2011).Effects of salinities on thegene expression of a NAD+-dependent glycerol-3-phosphate dehydrogenase inDunaliella salina.Sci Total Environ409,1291-1297.
5.Gee,R.,Goyal,A.,Byerrum,R.U.,and Tolbert,N.E.(1989).Twoisozymes of dihydroxyacetone phosphate reductase in Dunaliella.PlantPhysiol91,345-351.
Claims (10)
1.CemGPDH蛋白，其氨基酸序列如SEQ ID NO:2所示。
2.编码根据权利要求1所述的CemGPDH蛋白的基因。
3.根据权利要求2所述的基因，其核酸序列如SEQ ID NO:1所示。
4.载体，其包含根据权利要求2或3所述的基因。
5.宿主细胞，其包含根据权利要求1所述的CemGPDH蛋白、根据权利要求2或3所述的基因或根据权利要求4所述的载体。
6.一种制备具有高棕榈油酸和/或油酸含量，或具有高总脂肪酸含量的酵母、植物或藻类的细胞的方法，所述方法包括将根据权利要求2或3所述的基因或根据权利要求4所述的载体转入所述酵母、植物或藻类的细胞中，所述酵母优选是酿酒酵母，更优选是尿嘧啶缺陷型酿酒酵母INVSC1；所述藻类优选是小球藻，更优选是椭圆小球藻(Chlorellaellipsoidea)；所述植物优选选自拟南芥、烟草、油菜、向日葵、大豆、番茄、蓖麻、棉花、芝麻或花生。
7.根据权利要求1所述的CemGPDH蛋白、根据权利要求2或3所述的基因或根据权利要求4所述的载体在制备具有高总脂肪酸含量的酵母、植物或藻类的细胞中的用途，所述酵母优选是酿酒酵母，更优选是尿嘧啶缺陷型酿酒酵母INVSC1；所述藻类优选是小球藻，更优选是椭圆小球藻(Chlorella ellipsoidea)；所述植物优选选自拟南芥、烟草、油菜、向日葵、大豆、番茄、蓖麻、棉花、芝麻或花生，所述具有高总脂肪酸含量的酵母、植物或藻类的细胞优选是具有高棕榈酸、棕榈油酸、硬脂酸和/或油酸含量的酵母、植物或藻类的细胞。
8.根据权利要求1所述的CemGPDH蛋白、根据权利要求2或3所述的基因或根据权利要求4所述的载体在提高酵母、植物或藻类的细胞中棕榈油酸和/或油酸含量，或总脂肪酸含量中的用途，所述酵母优选是酿酒酵母，更优选是尿嘧啶缺陷型酿酒酵母INVSC1；所述藻类优选是小球藻，更优选是椭圆小球藻(Chlorella ellipsoidea)；所述植物优选选自拟南芥、烟草、油菜、向日葵、大豆、番茄、蓖麻、棉花、芝麻或花生。
9.利用根据权利要求6所述的方法制备的具有高棕榈油酸和/或油酸含量，或具有高总脂肪酸含量的酵母、植物或藻类的细胞。
10.根据权利要求9所述的酵母、植物或藻类的细胞在生产生物柴油中的用途。
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