CN109666705A - 酿酒酵母菌中染色质重塑因子基因在提高发酵酒精产率的应用 - Google Patents
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Abstract
本发明公开了一种酿酒酵母菌中染色质重塑因子基因在提高发酵酒精产率的应用。染色质重塑因子基因包括SPT10,SWI3,NGG1,UME6和RSC1基因，其核苷酸序列如SEQ ID NO.1‑5所示。本发明为提高酒精发酵的生产效率提供了新的方法，为构建高产酒精的优良酿酒酵母工程菌株提供了重要的信息。
Description
技术领域
本发明属于生物能源技术开发技术领域，具体地说，涉及一种酿酒酵母菌中染色质重塑因子基因在提高发酵酒精产率的应用。
背景技术
微生物发酵法生产燃料酒精是当前研究的热点，酿酒酵母菌(Saccharomycescerevisiae)是目前最为理想的酒精发酵生产菌株。目前国内外利用玉米淀粉作为原料，通过酿酒酵母菌发酵生产燃料酒精的技术已经非常成熟。通过传统的菌种选育手段(如自然筛选、诱变育种)来提高菌株的酒精发酵产量，已经基本到了极限。近年来，随着分子生物学和组学技术的快速发展，为大规模研究酿酒酵母基因和酒精发酵的关系提供了条件。通过染色质重塑，可以同时改变许多基因的表达水平，从而获得新的性状，是通过单个基因的改变所不能比拟的。
在现有技术中还没有通过染色质重塑方法来提高酿酒酵母酒精产率的报道。
发明内容
有鉴于此，本发明针对上述的问题，提供了一种酿酒酵母菌中染色质重塑因子基因在提高发酵酒精产率的应用。
为了解决上述技术问题，本发明公开了一种酿酒酵母菌中染色质重塑因子基因在提高发酵酒精产率的应用。
可选地，包括以下步骤：在YPD平板上划线活化含有染色质重塑因子基因的酿酒酵母菌，30℃培养2天；取一个单菌落接种30mL的YPD液体培养基，30℃，220r/min摇床培养16小时以上至饱和；按10％的比例把种子菌液接种到100mL的发酵培养基中，30℃，220r/min进行摇床培养8小时，然后转为静置培养发酵。
可选地，所述的YPD液体培养基含葡萄糖2％，酵母提取物1％，蛋白胨2％，去离子水溶解，pH7.0，通过高压灭菌，灭菌条件为115℃，20min。
可选地，所述的发酵培养基中各种成分的含量(g/L)为：葡萄糖100，硫酸铵7.5，磷酸二氢钾3.5，七水硫酸镁0.75，酵母提取物0.2，组氨酸0.02，尿嘧啶0.02，亮氨酸0.1，pH7.0，通过高压灭菌，灭菌条件为115℃，20min。
可选地，所述的染色质重塑因子基因，其特征在于，包括SPT10,SWI3,NGG1,UME6和RSC1基因。
可选地，其核苷酸序列如SEQ ID NO.1-5所示。
与现有技术相比，本发明可以获得包括以下技术效果：
1)本发明通过功能基因组学技术筛选，发现5个染色质重塑因子基因(SPT10,SWI3,NGG1,UME6或者RSC1)的缺失，能够提高酿酒酵母菌发酵的酒精得率。
2)本发明为提高酒精发酵的生产效率提供了新的方法，为构建高产酒精的优良酿酒酵母工程菌株提供了重要的信息。
当然，实施本发明的任一产品并不一定需要同时达到以上所述的所有技术效果。
附图说明
此处所说明的附图用来提供对本发明的进一步理解，构成本发明的一部分，本发明的示意性实施例及其说明用于解释本发明，并不构成对本发明的不当限定。在附图中：
图1是本发明五个染色质重塑因子基因(SPT10,SWI3,NGG1,UME6或者RSC1)的缺失菌株和野生型酿酒酵母菌株(WT)在发酵培养基中培养32小时(32h)和52小时(52h)后的产率(酒精产量/菌体干重比)；其中，*号表示每个基因缺失菌株和野生型菌株分别在32小时和52小时时间点的比较存在显著性差异。
具体实施方式
以下将配合实施例来详细说明本发明的实施方式，藉此对本发明如何应用技术手段来解决技术问题并达成技术功效的实现过程能充分理解并据以实施。
实施例1SPT10,SWI3,NGG1,UME6和RSC1的筛选方法
从双倍体酿酒酵母基因缺失株文库(美国Invitrogen Inc公司)中，挑选出150个与基因转录相关的基因的单基因缺失菌株，按照下面实施例2的方法，对它们进行酒精发酵实验，发现SPT10,SWI3,NGG1,UME6和RSC1这五个染色质重塑因子基因的缺失株菌株比野生型菌株的酒精产率都显著提高。SPT10,SWI3,NGG1,UME6和RSC1的基因序列如SEQ ID NO.1-5所示。
实施例2酿酒酵母菌的酒精发酵
在YPD平板上划线活化实验菌株，30℃培养2天。每个菌株取一个单菌落接种30mL的YPD液体培养基，30℃，220r/min摇床培养16小时以上至饱和。按10％的比例把种子菌液接种到100mL的发酵培养基中，30℃，220r/min进行摇床培养8小时，然后转为静置培养发酵。
其中，种子培养基(YPD)含葡萄糖2％，酵母提取物1％，蛋白胨2％，去离子水溶解，pH7.0。发酵培养基中各种成分的含量(g/L)为：葡萄糖100，硫酸铵7.5，磷酸二氢钾3.5，七水硫酸镁0.75，酵母提取物0.2，组氨酸0.02，尿嘧啶0.02，亮氨酸0.1，pH7.0。两种培养基均通过高压灭菌(115℃，20min)。
在接种后32小时和52小时两个发酵时间点，取1mL的菌体发酵液，测定菌体干重。另外取1mL的菌体发酵液，12000rpm离心5分钟，上清液取出用于液相测定其中的酒精浓度。将测得数据做成Excel表，并作出菌体干重、酒精产量、酒精产率图。
酒精含量的测定方法：用高效液相色谱仪(HPLC)检测发酵液中酒精的含量，利用示差折光检测器进行检测。所用色谱柱是BIO-RAD有机酸柱，柱温为35℃，流动相是0.0275％(v/v)稀硫酸，流速为0.6mL/min。每个样品进样量为20μL，检测时间是25min。
酒精对菌体得率按如下的公式计算：
式中y为酒精产率，p为酒精浓度，x为OD值。
如图1所示。经过52小时发酵，相比于野生型菌株(WT)，SPT10、SWI3、NGG1、UME6和RSC1五个单基因缺失酿酒酵母菌株的酒精发酵产率，分别提高了35.0％、37.0％、36.0％、50％和37.5％。
上述说明示出并描述了发明的若干优选实施例，但如前所述，应当理解发明并非局限于本文所披露的形式，不应看作是对其他实施例的排除，而可用于各种其他组合、修改和环境，并能够在本文所述发明构想范围内，通过上述教导或相关领域的技术或知识进行改动。而本领域人员所进行的改动和变化不脱离发明的精神和范围，则都应在发明所附权利要求的保护范围内。
序列表
<110> 山东理工大学
<120> 酿酒酵母菌中染色质重塑因子基因在提高发酵酒精产率的应用
<130> 2018
<160> 5
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<211> 1923
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<400> 1
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tttaggcgga ttggaaagct gcctgaagcc gggattctca agggctttga tgtcccagta 780
gattctttca tgtatggtaa agagttcaca catattacga aaagtataga tttgttacga 840
gatcctcaaa agagtatcga aattgggaaa tatgaacgat taaagcattt tctggaaaca 900
ggcaagtatc ctttacattg tgatagaaac gaaaaggcta gattaagagt actttccaaa 960
actcattcgg tattgaatgg caagttgatg acaaaaggaa aagaaatcat ttatgacacg 1020
gatcaacaaa tccagattgc attagagata catttaatgg aacatttagg catcaacaag 1080
gtgacctcca aaattggtga aaaataccat tggagaggaa tcaagagtac tgtttcggag 1140
gtaatctctc ggtgccaaaa atgcaagatg aggtacaaag acgggacagg ggtaataatt 1200
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aacaatccaa gaaaaagcaa gaagcatgat aatgcgttat tgggacaggc aataaatttc 1320
cctcaaaata ttatttccag tacgctgaat gatgtggagg gtgaacccac tccgccagac 1380
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atgcaagcga ataagctaga ttcccaggaa acaccttcta cagaagaaag cagggcccag 300
aacgtctttg gccaagataa tgaggatagt gataatctct ttggcgaaac cgagagttct 360
gtttcgaaca atgaggcaaa cactcctagt atacctacga acccggtaga caatgaaaac 420
aataagcctg ctattaagga agacagcaca atacaggatt caaatggtga cgttaaaaac 480
atggaggacg tcaaaataca aaaagaggaa gaaccagaga ataacactgt tattgaaggt 540
gtcaaagaag aaagtcagcc agatgagaac acaaaggaaa tggatgaagt ggaagaagat 600
gatgaggatg acgatcaacc gatgattagt cctgataatt ccatttttgg agacacaaag 660
agtgagtcta aacaactagg caacacttcg tcggttgcta atactccatc tgaaattcca 720
gatgcgcata aagctgaaca ggaggatatc atagaaaaga cggaaagtgt agataagaag 780
gtagatagtg gcgaagagag aaacgaacag gaacgtgaaa tcatgaacga tcactccaaa 840
agtgccaatc caaagaaaac cactatcacc agagtagaac ccgaaacttt tgagataccg 900
caagcacatg aaattgttat tccaagttat tctaaatggt tcaatctaga aaaaattcat 960
tccattgaag ttcaatcgtt acctgaattt ttcaccaata ggattccatc aaagacacct 1020
gaagtttaca tgagatacag aaacttcatg gtaaattctt acaggctgaa ccccaacgaa 1080
tatttcagtg tcactacggc gaggagaaac gtctctggtg atgctgctgc attattcaga 1140
ttgcataaat tcttaacaaa atgggggcta ataaactatc aggttgactc caaactatta 1200
cctaagaaca tcgaaccccc actaacgtct caatattcca caaggcatga tgcccccaga 1260
ggactattcc cattcgaaag ttacaaacca tctgtacaac tgccggatat ggcaaagctt 1320
aaaaaaatga tgaacactag tgattcggag agtactttat ataaatattt gaaagaaagc 1380
aaaagaaaat atgatgaaat aactcacccc cccagtacca ctgatgatga aaatggtgac 1440
aaaaatgata atggtggtaa aatgaacaat gaagtaagta cgtcaacaag catgactggt 1500
gatgccaatc tattggaaga aggcgaaact tcacgtcctc tgaaaaaagt caaaattcta 1560
gaacaaattg atgaaaattg gtcaaaagaa gatttgcaaa agctattaaa gggaatacaa 1620
gaattcggcg ctgactggta caaagtggcc aaaaacgttg gcaataagtc gcctgaacag 1680
tgtatcttac gtttccttca attgcccatt gaagataaat ttttatacgg tgacggcaat 1740
ggcaagggtg ataatgacaa cgggttagga cctttgaaat atgctcctca cttgccattt 1800
tcgaagagcg aaaaccccgt tttatcaaca atagcattct tggtgggttt ggtcaatccg 1860
aagactgtac agtcgatgac ccagagagcc attcagtccg cagaatcaat aaaatcacaa 1920
aaagaggaaa tctctgacca aaagccaatt gaacatatta aagaaggttc agaaattgcc 1980
atatcatcat tgggttatag gagtcatatt ttcgctacca acgaggaaag gcaaatgaat 2040
tttttgacta acgagctaat aagacttcaa atggaaaaat tggacgctaa attaaatcat 2100
ttaaagaaac tagaaaaatt tatggagcta gaaaggaaaa ccctagaaag acaacaggaa 2160
aacttactaa ttcagagatt gaacttcaac caaaactcca gcaagattgt aaacgtttta 2220
agcaaatgct tgaatttaat atccgacagc aatatcaata atagttcagt ggcggagaag 2280
gaggaaatca ggtcgcagat agaccatttc aaaagtatgc tatcgaaacc tgaaacttta 2340
agtattggaa agaatccatt taacaaacct aatattgaaa caggtgaaaa ccataatggc 2400
caaagtatat ctaacgagaa tgatgtcaaa cccatttcta ttgaagcacc tcaattttac 2460
aggtattggt cggcataa 2478
<210> 3
<211> 2511
<212> DNA
<213> 酿酒酵母菌(Saccharomyces cerevisiae)
<400> 3
atgctagaca aggcgcgctc tcaaagcaaa cacatggacg aatctaatgc ggctgcctct 60
ctgctttcga tggaaacaac cgccaacaat catcactatt tgcacaataa aacatctcgt 120
gccacgctga tgaatagcag ccaagacggc aaaaaacatg cagaagatga agttagtgat 180
ggagctaact cccgccatcc tacaatttcc agtgctagca tcgaatctct caagacaacc 240
tacgatgaaa accctttgct ttctattatg aaatcgacat gtgcgcccaa caacactccc 300
gtgcatactc cgtctggttc gccgagtttg aaagtccaaa gtggcggaga tatcaaagac 360
gatcctaagg aaaacgatac tactactacg accaatacta cacttcaaga tcgccgtgac 420
agcgataacg ctgtgcatgc tgccgccagc ccactcgcgc cttccaatac accttcggat 480
cctaagtcat tgtgcaatgg ccatgtcgca caggctacag acccacaaat ttccggtgct 540
attcagccgc agtatactgc aaccaacgag gatgttttcc cttactcctc cacctccact 600
aatagtaaca ctgccactac tactatcgtc gccggcgcca aaaaaaaaat acatttgccg 660
ccaccacaag ctccagcggt ttcttccccc ggtaccaccg cagcaggctc gggcgcgggc 720
acgggctcgg gcatccgttc ccgcacagga tcggatttgc cgctcatcat taccagcgcc 780
aacaagaaca acggtaagac taccaattcg cctatgtcga tactgagcag aaacaacagt 840
accaacaaca acgacaataa ttcgatacaa agctcagatt cgagagaatc ctctaataac 900
aatgagattg gcgggtattt gcgcggcgga actaagcgcg gcggcagtcc atctaacgac 960
tctcaggtcc agcataatgt gcatgatgac caatgtgccg tgggcgtggc gcccaggaac 1020
ttctatttca acaaggatag agagataaca gacccaaatg taaaactgga cgagaacgaa 1080
tcaaaaatca acatatcgtt ctggctaaat tcgaaataca gagatgaggc ttattctttg 1140
aatgaatcat cctccaacaa tgctagttca aacacggata cgcctacaaa ctctcgacat 1200
gcgaacacca gctcttccat taccagcaga aacaatttcc agcattttag gttcaaccaa 1260
ataccttctc aacctccaac ttccgcatct tcgtttacaa gcacaaacaa caacaaccct 1320
caacggaaca atatcaatcg cggtgaagac ccgtttgcca cttcgtcaag accttctact 1380
gggttttttt acggcgattt gccgaatcgt aacaatagaa atagtccctt ccatacaaat 1440
gaacaataca tcccaccacc tccaccgaaa tacatcaatt ctaaattgga tggattgaga 1500
tcaagattat tgctcggtcc gaattctgca tcttcatcta ccaaactaga cgacgacttg 1560
ggtacagcag cagcagtgct atcaaacatg agatcatccc catatagaac tcatgataaa 1620
cccatttcca atgtcaatga catgaataac acaaatgcgc tcggtgtgcc ggctagtagg 1680
cctcattcgt catcttttcc atcaaagggt gtcttaagac caattctgtt acgtatccat 1740
aattccgaac aacaacccat tttcgaaagc aacaattcta cagctgtttt tgatgaagac 1800
caggaccaaa atcaagactt gtctccttac catttaaatc taaactctaa aaaggtttta 1860
gatcccactt ttgagtcaag gacaaggcaa gttacttgga ataagaatgg taagcgaata 1920
gacagacgcc tttctgctcc agaacaacaa cagcaactgg aagttccacc attgaaaaaa 1980
tcgagaaggt cagttggaaa cgcaagagta gcaagccaaa ccaatagcga ttataattct 2040
cttggcgaat cctcaacttc gtcagctcca tcgtctccat ctttgaaggc ttcttctggc 2100
ttggcatata ccgctgatta tcctaacgct acttcgccgg atttcgctaa atctaaagga 2160
aaaaatgtca agcctaaggc aaaatcaaag gcgaaacagt catcaaagaa aagaccaaat 2220
aatactactt cgaaatcaaa agcaaacaat tctcaagaat cgaataatgc tacttcctca 2280
acgtctcaag gtacaaggtc ccgtactggt tgctggattt gtagattaag gaaaaagaag 2340
tgtaccgagg aaagaccgca ctgtttcaac tgtgaaaggt tgaaattgga ctgtcactat 2400
gacgcgttca aaccagattt tgtatctgat ccaaagaaaa aacagatgaa actggaggaa 2460
atcaagaaaa aaacaaaaga ggccaaaaga agagcaatga aaaaaaaata a 2511
<210> 4
<211> 2109
<212> DNA
<213> 酿酒酵母菌(Saccharomyces cerevisiae)
<400> 4
atgcctagac atggaagaag aggaaaactg ccaaaaggcg agaaacttcc taaaaaggag 60
ggaggcgaca atactccgtc aaagctttta tcgtcaatgc ttaaaacttt agatttgacc 120
ttcgagcgtg atattggaat gctcaatggc aaaagtgtta ggtccatacc caataaaaaa 180
actttacttg aattgcaaag tcaattggat agtctcaatg aaatactggg tacaattgcc 240
cgaggtgatc aagagaccat tgaagcgctt cggaaaataa gagacagtaa aaatgagaag 300
caagcaaacg atgaaaagca ggaaacttca aatgcagacg ggcagcacga aagttctact 360
gctactgaag aaacgaatat aattgataaa ggggtacaaa gcccaccaaa acctcctcct 420
tctaatgaaa tctccggcac aattgagaac gatgtagaaa gcatcaaaca agcggctgat 480
aatatggcta aagaagaaat taatgaagac aaagatttac aagttcatag agatcaaccc 540
agagaaaaga ggccgttcga ttctgaaact gaaaataggg ccactgaaaa tgaaaacaca 600
cagaggccag acaataagaa gcaaaaaatt gacgttgaca aaatggaaaa cgatccaact 660
gttaaaaatc caaaatcaga atttgtagta tcgcaaacgt tacctcgagc agcagcagca 720
ctcggcttat tcaatgaaga gggactagaa agtacaggag aagacttcct aaaaaaaaaa 780
tacaatgtag ctagttatcc aacaaatgat ttgaaagatc tgttaccagg tgaattaccg 840
gatatggact tttcgcatcc taaaccaacc aaccaaattc aattcaatac ctttctagca 900
tttgtggaaa attttttcaa agatcttagt gatgacaact taaagttttt gaagatgaaa 960
tatattatac ccgatagctt gcaatttgac aaaacttatg accctgaggt aaacccgttt 1020
attataccga aactaggccc tttatacaca gatgtttggt tcaaagatga aaacgacaaa 1080
aattctgcct ataaaaaacc ttcaccatat tcaaacgatg catctactat actaccaaaa 1140
aaaagtgcca acgaacttga tgataatgct ttggaatcgg gcagtatatc gtgtgggccc 1200
ttattatcta ggctgttgag tgctgtatta aaagatgaca atgacaaatc agaattgcaa 1260
tcttctaaaa taatacggga tggcggacta ccgagaaccg gaggggaaga cgatatacaa 1320
tcgtttagaa acaacaacaa tgatactgta gacatgacat tatctcagga gaacgggccc 1380
agtgttcaaa cacctgacaa tgacattgat gaggaagcat ctttccaagc aaaactggca 1440
gaaaataaag gcagtaatgg cggtactaca agtacgttac cccaacaaat tgggtggata 1500
acaaatggaa ttaacctgga ctatccaaca ttcgaagaac gattgaaaag agaactaaag 1560
tatgtaggga tatacatgaa tttgcctaag gatgagaaca accctaactc agatgacccc 1620
gattgggtca ccggtagaga agacgacgaa ataagcgcag agttaagaga attgcaaggc 1680
actttaaaac aagtgaccaa gaaaaaccaa aagaggaaag cccaattgat tccactagtg 1740
gaaagacaac tagcatggca ggaatactca tctattttag aagatttaga caaacaaatc 1800
gaccaggctt atgtcaaacg tattcgcgta cccaagaaaa gaaagaagca tcatacagca 1860
gcttcaaata atgtgaacac aggaaccacg tctcaaatag cacaacaaaa ggctgcaaac 1920
tcaagtttaa aatcccttct ggataaaagg caaaggtgga ttaataagat tggtccattg 1980
tttgataaac ctgaaattat gaagagaatc cccaatgaaa gcgtattcaa ggacatggac 2040
caagaagaag atgaggatga agccgatgta tttgcacaaa acactaataa ggacgtggaa 2100
ctaaattaa 2109
<210> 5
<211> 2787
<212> DNA
<213> 酿酒酵母菌(Saccharomyces cerevisiae)
<400> 5
atggtggagc aagataatgg gtttttacag aagctactca aaacacaata cgatgctgtc 60
ttccatttaa aggatgaaaa tggtatagaa atatatccaa tattcaatgt actgccacct 120
aaaaaagaat atccagacta ttacattata ataaggaatc cgatatcttt aaacacttta 180
aagaaacgat tgccccatta tacatcgcca caagattttg ttaacgattt tgcgcaaatt 240
ccttggaatg caatgacata caatgcaaaa gattcagtaa tttacaaata tgccattctt 300
cttgagtctt ttattaaagg aaagatcgtg cacaatataa gaaaacatta tcccgaggtg 360
acatatccat ctctaggtcg aattccggaa atattcgcag agagtatgca accctccgat 420
ttatcctcga accccattaa tacacaggaa aatgatgaaa aggctggctt aaatccagag 480
atgaaaatgg ctttcgcaaa actagatagt tcgatcactg aaaggaaacc aaccaaccaa 540
gattacagga tgcaacaaaa aaactcacca gcgtttccaa cccattctgc ttctataaca 600
ccgcagccgc tggcttcacc tactccagtt gttaactatg caaatattac ctcggcgcat 660
cccaaaaccc atgttagacg cggtagacca ccagtaattg atctacccta tgtactgagg 720
attaagaata tcttgaagat gatgagaaga gaggtcgacc aaaataataa gacgcttacc 780
ctctgttttg aaaaactacc agatagaaac gaggagccaa cctattactc tgtcataaca 840
gatcccattt gtctaatgga tattaggaaa aaggttaagt ccagaaaata tagaaatttc 900
catacttttg aggaagactt tcaattaatg ctaacaaatt ttaagcttta ttattctcaa 960
gatcagagca atataatacg agctcaacta ttggagaaaa acttcaatag attagtgcga 1020
attgaattat ccaaacctga tgaagactat ttacctgaag gtgaattaag atatcctcta 1080
gatgatgtag aaatcaatga tgagaaatac cagataggtg attgggttct tttacataat 1140
ccaaacgaca tcaataaacc cattgttggc cagattttta gattatggtc tactacagat 1200
ggaaacaaat ggttgaatgc ttgctggtat tttaggccag aacaaaccgt tcatcgggtg 1260
gataggttat tctacaagaa tgaggtcatg aaaactggtc aatatagaga tcatcctatt 1320
caggatatta agggaaagtg ctacgttatt cattttacca gattccagcg tggggatcct 1380
agcacgaaag tcaatggtcc tcaatttgtt tgcgaattcc gttataatga aagtgataag 1440
gttttcaaca aaatcaggac ttggaaagct tgcctgccgg aagaactccg tgaccaagat 1500
gagcctacga tacctgttaa tggaagaaaa ttttttaaat acccttctcc aatagcagat 1560
ttgttaccag caaacgcaac tctaaatgac aaagttccag aacccacgga gggagcccca 1620
actgccccac ctttggtagg cgctgtatat ttaggcccca aattagagag ggatgatctg 1680
ggagagtatt caacttctga tgactgccct cgatacataa tcagaccaaa tgatccacca 1740
gaagaaggta aaatagatta cgaaacagga acaataatta cagatacctt aaccacaagc 1800
agtatgccca gagtgaacag ttcctccaca atacgtttac ccactttaaa gcagactaaa 1860
tcaattccca gctccaattt caggtcctca tcaaatactc cactattgca tcagaacttc 1920
aatcagactt ccaatttttt gaaattagaa aacatgaata atagttctca taatttacta 1980
tcacatcctt ccgttcctaa gtttcaatcg ccctctttat tagaacagag tagtcgctct 2040
aaataccata gtgcaaaaaa acagacacag ctatcatcaa cagctcccaa aaagcctgca 2100
tctaaaagtt ttacattatc ctcaatgatt aataccttaa ctgctcatac ctcgaagtac 2160
aacttcaacc atattgttat tgaagccccg ggtgcatttg tggttcctgt accaatggaa 2220
aaaaatataa gaaccatcca atcgacagaa cgtttcagta gatcaaatct aaaaaacgct 2280
caaaatctcg gtaatactgc tattaatgac ataaatactg caaatgagca aataatatgg 2340
ttcaaagggc ccggtgtaaa aatcacagag agagttattg attccggaaa tgatttagta 2400
cgggtacctt tgaatagatg gttctgtaaa aacaaaagga gaaaactgga ttacgaagac 2460
attgaagaag atgttatgga accaccaaat gacttcagcg aagatatgat tgctaatatt 2520
tttaatcctc cgccgagctt gaatttagat atggatttaa atctaagccc atcctcgaat 2580
aactcttcta atttcatgga tttaagcaca attgcaagcg gtgataacga tgggaaagaa 2640
tgcgatacgg cggaggagag tgaagacgaa aacgaagata cagaagacga gcacgaaatt 2700
gaagatatcc cgaccacgtc tgcttttggc ttaaattctt ccgctgaata ccttgcattc 2760
agattacgag aatttaataa attatga 2787
Claims (6)
1.酿酒酵母菌中染色质重塑因子基因在提高发酵酒精产率的应用。
2.根据权利要求1所述的应用，其特征在于，包括以下步骤：在YPD平板上划线活化含有染色质重塑因子基因的酿酒酵母菌，30℃培养2天；取一个单菌落接种30mL的YPD液体培养基，30℃，220r/min摇床培养16小时以上至饱和；按10％的比例把种子菌液接种到100mL的发酵培养基中，30℃，220r/min进行摇床培养8小时，然后转为静置培养发酵。
3.根据权利要求2所述的应用，其特征在于，所述的YPD液体培养基含葡萄糖2％，酵母提取物1％，蛋白胨2％，去离子水溶解，pH7.0，通过高压灭菌，灭菌条件为115℃，20min。
4.根据权利要求2所述的应用，其特征在于，所述的发酵培养基中各种成分的含量(g/L)为：葡萄糖100，硫酸铵7.5，磷酸二氢钾3.5，七水硫酸镁0.75，酵母提取物0.2，组氨酸0.02，尿嘧啶0.02，亮氨酸0.1，pH7.0，通过高压灭菌，灭菌条件为115℃，20min。
5.根据权利要求2所述的应用，其特征在于，所述的染色质重塑因子基因，其特征在于，包括SPT10,SWI3,NGG1,UME6和RSC1基因。
6.根据权利要求5所述的基因，其特征在于，其核苷酸序列如SEQ ID NO.1-5所示。
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| WO2004079008A1 (en) * | 2003-03-04 | 2004-09-16 | Suntory Limited | Screening method for genes of brewing yeast |
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| WO2014105840A1 (en) * | 2012-12-31 | 2014-07-03 | Butamax Advanced Biofuels Llc | Fermentative production of alcohols |
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| EP2446017A2 (en) * | 2009-06-26 | 2012-05-02 | Archer Daniels Midland Co. | Improvement of ethanol yield and reduction of biomass accumulation in the recombinant strain of saccharomyces cerevisiae overexpressing atp degrading the enzymes |
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Application publication date: 20190423 |