CN109666683A - 乙酰辅酶A乙酰转移酶基因RKAcaT2及其应用 - Google Patents
乙酰辅酶A乙酰转移酶基因RKAcaT2及其应用 Download PDFInfo
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- CN109666683A CN109666683A CN201910147204.0A CN201910147204A CN109666683A CN 109666683 A CN109666683 A CN 109666683A CN 201910147204 A CN201910147204 A CN 201910147204A CN 109666683 A CN109666683 A CN 109666683A
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Abstract
本发明公开了一种乙酰辅酶A乙酰转移酶基因RKAcaT2及其用途,其核苷酸序列如SEQ ID NO:1所示,该基因编码的氨基酸序列如SEQ ID NO:2所示,该基因是红冬孢酵母(Rhodosporidium kratochvilovae)YM25235中类胡萝卜素合成的关键酶基因,具有乙酰辅酶A乙酰转移酶的功能,能够调控红冬孢酵母YM25235产类胡萝卜素;通过基因工程手段对微生物进行改造,来提高微生物体内类胡萝卜素的产量,为大规模商业化生产类胡萝卜素奠定基础。
Description
技术领域
本发明属于生物技术领域和遗传工程领域,涉及一种乙酰辅酶A乙酰转移酶基因RKAcaT2,具体涉及从酵母---红冬孢酵母(Rhodosporidium kratochvilovae)YM25235中克隆的乙酰辅酶A乙酰转移酶基因RKAcaT2以及将该基因直接与不同载体连接,转入酵母细胞,来提高这个基因的表达水平并最终促进类胡萝卜素的合成。
背景技术
类胡萝卜素(carotenoid)是所有光合生物及一些非光合原核生物和真菌合成的亲脂性天然色素,在自然界中广泛存在,一般呈黄色、橙红色、红色或紫色,典型的类胡萝卜素是由8个异戊二烯单元首尾相连而成的C40萜类化合物及其衍生物,其结构中的共轭双键的作用导致其各种类胡萝卜素的颜色差异。一些类胡萝卜素侧碳链较短(C30)或较长(C45或C50),根据结构中是否含有氧官能团分为碳氢类胡萝卜素和含氧衍生物叶黄素。由于其分子结构中类异戊二烯宫二结构的存在,吸光性较强,所以一般在波长430~570nm之间有吸收峰。到目前为止,自然界已有700多个种类的类胡萝卜素被发现。
天然类胡萝卜素对于生物体具有十分广泛的作用。首先,天然类胡萝卜素具有超氧自由基阴离子介导的抗氧化性质;防紫外线的光保护性能;调节细胞分化、细胞周期和凋亡等。在工业上类胡萝卜素被用作食品着色剂、营养强化剂和抗氧化剂等。叶黄素可作为食品着色剂用于面条、色拉调味品等食品中;在一些调味品中,利用番茄红素具有超强的抗氧化活性,代替亚硝酸盐防止食物腐败变质,有效的延长了保质期。由于大部分动物不能合成类胡萝卜素,需要从食物中获取,且人工合成的类胡萝卜素不具有这些特殊的生理功能,因而天然类胡萝卜素的制备相当重要。
随着对天然类胡萝卜素制品的需求增加,尽管已有许多研究者对如何提高类胡萝卜素的产量进行了相当多的研究,但现在类胡萝卜素的产量和质量仍不能满足市场需求。目前,类胡萝卜素的生产方法主要包括植物提取法、化学合成法和生物合成法。化学合成法生产类胡萝卜素相较于其他方法而言,是成本最低且简便的,但由于存在生物活性低且生成色素有一定的毒副作用,已被许多国家和地区限制使用;植物提取法由于植物中类胡萝卜素含量较低,提取成本比较高、工艺又复杂等原因,使用也较少;而生物合成法提取的类胡萝卜素是纯天然的,对人体安全无毒副作用,因此用生物合成法提取类胡萝卜素是目前最具发展潜力的一种提取方法。微生物发酵生产类胡萝卜素只需要低成本天然底物作为碳源,该方法还具有生物培养周期短、产量高等特点,且微生物发酵生产类胡萝卜素不存在由于季节性和地域性变化而导致的产量和市场问题,目前来说,能够发酵产生类胡萝卜素的微生物主要有真菌、细菌和酵母菌等,其中利用酵母发酵生产类胡萝卜素的研究最为常见,且酵母属于我国饲料行业中确认的饲用微生物。因此,用微生物发酵法来替代其他方法可以从一定程度上缓解天然类胡萝卜素的生产发展局限,该方法将有助于实现发酵生产类胡萝卜素产品的工业化。
发明内容
本发明目的是提供一种乙酰辅酶A乙酰转移酶基因RKAcaT2,该基因是从红冬孢酵母(Rhodosporidium kratochvilovae)YM25235中分离得到,该基因核苷酸序列如SEQ IDNO:1所示或该核苷酸序列的片段,或与SEQ ID NO:1互补的核苷酸序列,该基因序列长为1191bp(碱基),该基因编码的氨基酸序列如SEQ ID NO:2所示的多肽或其片段。
本发明另一目的是提供一种含有乙酰辅酶A乙酰转移酶基因RKAcaT2的重组表达载体,是将SEQ ID NO:1所示基因直接与不同表达载体(质粒、病毒或运载体)连接所构建的重组载体。可用本领域的技术人员熟知的方法来构建含乙酰辅酶A乙酰转移酶基因RKAcaT2的核苷酸序列和合适的转录/翻译调控元件的表达载体;这些方法包括体外重组DNA技术、DNA合成技术、体内重组技术等;所述乙酰辅酶A乙酰转移酶基因RKAcaT2的核苷酸序列可有效连接到表达载体的恰当启动子上,以指导mRNA合成。这些启动子的代表性例子有:大肠杆菌的lac或trp启动子;λ噬菌体的PL启动子;真核启动子包括CMV早期启动子、HSV胸苷激酶启动子、早期和晚期SV40启动子、反转录病毒的LTRs和其它一些已知的可控制基因在原核细胞或真核细胞或其病毒中表达的启动子。表达载体还包括翻译起始用的核糖体结合位点和转录终止子等。在载体中插入增强子序列将会使其在高等真核细胞中的转录得到增强。增强子是DNA表达的顺式作用因子,通常大约有10-300bp,作用于启动子以增强基因的转录。如腺病毒增强子。
本发明另一目的是提供一种含有乙酰辅酶A乙酰转移酶基因RKAcaT2或上述重组表达载体的宿主细胞。
本发明的DNA片段序列也能用下列方法获得:(1)从基因组DNA分离双链DNA序列;(2)化学合成DNA序列以获得所述多肽的双链DNA。
本发明另一目的是上述乙酰辅酶A乙酰转移酶基因RKAcaT2应用在产类胡萝卜素中。
本发明从红冬孢酵母(Rhodosporidium kratochvilovae)YM25235的总RNA基因中分离得到乙酰辅酶A乙酰转移酶基因RKAcaT2,该基因全长1191bp;红冬孢酵母YM25235中RKAcaT2基因的过表达会引起细胞内这个基因的转录水平在一定程度上有一定的提高,说明外源基因在菌体内发生转录,继而翻译成相应蛋白,引起细胞内与类胡箩卜素合成相关的酶的表达量的提高。本研究结果有助于阐明红冬孢酵母YM25235中产类胡萝卜素机制,为揭示微生物提高类胡萝卜素产量机制提供参考,将有助于通过基因工程手段对其进行改造来提高类胡萝卜素含量,对类胡萝卜素的工业化生产提供有良好的应用前景和经济效益,为大规模商业化生产类胡萝卜素奠定基础。
附图说明
图1为本发明的红冬孢酵母YM25235的RKAcaT2基因PCR扩增图;
图2为重组质粒pRHRKAcaT2的质粒图谱;
图3为重组质粒pRHRKAcaT2限制性酶切分析;其中:1、DNA 分子量标记DL10000 2、空质粒pRH2034的NocⅠ、EcoRⅤ双酶切;3、重组质粒pRHRKAcaT2的NocⅠ、EcoRⅤ双酶切;4、RKAcaT2基因的PCR 产物;5、DNA 分子量标记DL2000;
图4 重组质粒pRHRKAcaT2转化红冬孢酵母YM25235阳性克隆验证;1.DNA分子量标记DL5000;2.野生型菌株特异性基因条带;3.重组菌株特异性基因条带;4.特异性基因的cDNA条带;5. DNA分子量标记DL2000;
图5为过表达菌株YM25235/pRHRKAcaT2与对照菌株YM25235/pRH2034的总类胡萝卜素含量。
具体实施方式
下面结合附图和实施例对本发明作进一步详细说明,但本发明保护范围不局限于所述内容,实施例中使用的试剂和方法,如无特殊说明,均采用常规试剂和使用常规方法。
实施例1:从红冬孢酵母(Rhodosporidium kratochvilovae)YM25235中分离乙酰辅酶A乙酰转移酶基因RKAcaT2的核苷酸序列
采用生工生物工程(上海)股份有限公司的UNlQ-10 柱式 Trizol 总RNA抽提试剂盒(产品编号: SK1321)提红冬孢酵母YM25235的总RNA ,然后按照TaKaRa公司试剂盒PrimeScript ®RT reagent Kit With gDNA Eraser(Perfect Real Time)进行反转录合成cDNA,取0.5μL为模板进行聚合酶链式反应,根据转录组测序中发现的RKAcaT2序列,设计特异性引物RKAcaT2-F和RKAcaT2-R(引物1和引物2),以上述得到的cDNA模板在PCR仪(BIOER公司)上进行PCR扩增,反应所用引物、组份和扩增条件如下:
引物1:RKAcaT2-F:5’-TCACCATGGCCGTCTTCATCGCG-3’(SEQ ID NO:3)
引物2:RKAcaT2-R:5’-CTTGATATCCTACACCCGCTCAAAGAG-3’(SEQ ID NO:4)
(CCATGG为NocⅠ酶切位点,GATATC为EcoR V酶切位点);
PCR扩增体系如下(50 μL):
;
扩增条件:94℃预变性5 min,再用94℃变性30 s,58℃退火30 s,72℃延伸1min15 s,进行30个循环,最后72℃彻底延伸10min,反应完后取产物2μL,然后在浓度为1%的琼脂糖凝胶中进行电泳分析,结果如图1所示,扩增得到大小约1200bp的片段,用琼脂糖凝胶DNA回收试剂盒(北京索莱宝科技有限公司)回收,把回收片段连接至到pMD-18T(TaKaRa公司产品)中,连接产物转化到用CaCl2法处理的大肠杆菌DH5α,在含氨苄青霉素(100 µg/mL)的LB固体平板上过夜培养,挑取平板上生长的白色菌落,通过菌落PCR验证阳性克隆。将验证阳性的克隆子接入LB液体培养基(含100 µg/mL氨苄青霉素)中过夜培养,用高纯质粒小量制备试剂盒(离心柱型)(北京百泰克生物技术有限公司)提取质粒,经测序(昆明硕擎生物科技有限公司),测序结果显示所扩增得到的片段大小为1191bp,命名为RKAcaT2,序列组成如SEQ ID NO:1所示的核苷酸序列。
实施例2:过表达载体pRHRKAcaT2的构建
以反转录的YM25235 cDNA为模板,用RKAcaT2-F 和RKAcaT2-R 作为引物扩增RKAcaT2的编码序列,获得的RKAcaT2片段大小约1200bp,将扩增获得的RKAcaT2片段经NocⅠ、EcoRⅤ两个限制性内切酶进行酶切后,连接到表达载体 pRH2034上获得重组质粒pRHRKAcaT2(图2)。将获得的重组质粒转入大肠杆菌 DH5α中扩增,再经菌落 PCR 验证后提取重组质粒,并用NocⅠ、EcoRⅤ对pRHRKAcaT2进行双酶切验证。结果表明,重组质粒pRHRKAcaT2经双酶切后产生了1.2kb 和10.7 kb左右的两个条带(图 3 第3泳道),这两个条带分别与RKAcaT2片段和pRH2034载体经双酶切后的片段大小一致,初步表明重组质粒pRHRKAcaT2构建成功;用测序引物进行测序,将酶切验证正确的质粒送出测序进一步进行验证;测序结果表明,测序所获得的序列与目的序列完全一致,没有出现任何碱基突变和缺失等。
实施例3:RKAcaT2基因过表达对红冬孢酵母YM25235中类胡萝卜素合成的影响
1、农杆菌介导转化红冬孢酵母YM25235
利用农杆菌介导法将重组质粒pRHRKAcaT2转化至红冬孢酵母YM25235中,以含潮霉素B(Hygromycin B)终浓度为150µg/mL的YPD 培养基筛选转化子,然后按照上海生工生物工程股份有限公司DNA 提取试剂盒说明书中步骤提取酵母转化子的基因组 DNA,后进行PCR验证,结果如图4所示。
2、RKAcaT2基因过表达的红冬孢酵母YM25235中类胡萝卜素含量分析
将含pRHRKAcaT2的过表达菌株YM25235/ pRHRKAcaT2在30℃条件下培养144h,提取类胡萝卜素,并以转入空质粒pRH2034的红冬孢酵母菌株YM25235/ pRH2304为对照,利用紫外-可见分光光度计在445nm下测定总类胡萝卜素的含量(mg/g干菌体),含量如图5所示;由图可看出,其中过表达菌株YM25235/pRHRKAcaT2的总类胡萝卜素合成量比含空质粒pRH2034的对照菌株YM25235/ pRH2304有明显提高,含空质粒pRH2034的对照菌株的类胡萝卜素合成量为3.77mg/g,而过表达菌株YM25235/pRHRKAcaT2类胡萝卜素合成量为6.52mg/g,即过表达菌株YM25235/pRHRKAcaT2类胡萝卜素合成量是对照菌的1.73倍,结果表明RKAcaT2基因能够促进总类胡萝卜素的合成,即RKAcaT2核酸序列确实与红冬孢酵母中类胡萝卜素合成有关。
序列表
<110> 昆明理工大学
<120> 乙酰辅酶A乙酰转移酶基因RKAcaT2及其应用
<160> 4
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1191
<212> DNA
<213> 红冬孢酵母(Rhodosporidium kratochvilovae)
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atggccgtct tcatcgcggc aggcaagcgc acggcgttcg gcgcgttcgg tggcgccctc 60
aagaactaca cggcctcgca gctcggcggg ttcgcggcca aggcggcgct cgcagagctt 120
cccgagggca cgcaggtcga ctcggtcatc ttcggccagg tcctctactc ggacccctcg 180
gccgcctacc tcgcacgcca cgtcggccac catgcgggcg ttcccgtgca tgtccctgcg 240
ctgaccgtca accgcctctg cgggagcggc ttccaggccg tcatcaacgc ggcgcaggag 300
atcaagctcg gcgagtcgca cgtcgtcctc accggcggca cggacaacat gtcgctctcg 360
ccctacacgc tctcgggctc gtctcgcttc ggcaacaagt acggcgtcga cctcaagctc 420
gaggactcgc tcgcgcaggc gctgacggac cgcgtgccta acccgacacc gatgggcatc 480
acggccgaga acttggcgca gaagtacggc atctcgcgcg agcagtgcga ccagtacgcg 540
ctgcagagcc agcagcggtg gaagaaggca ctcgactcgg gcgccttcga agccgagatc 600
acgccggtgc agctcccacc gaagaagcgc ggcggcgagc cgatcacctt ctcgcaggac 660
gagcacccgc gcccgcaggc gacgctcgag cagctcggcc gcctccccgc cgtctttgca 720
aagaacggca ccgtcactgc tggtaacgcg agcggaatct gcgacggcgc ggcggcgaac 780
gttgttgtca gcgaggaggc cgtcaagcgc tatgggctga agcccctcgc gagggtcgtg 840
gcttaccaca tcaacgcggt cgacccgaac attatgggca tcggccccgt cgagggtatc 900
cgcggcgtcc tgaagaaggc gggcatgaag attgaggaca tcgacctctt cgacatcaac 960
gaggcgttcg cggcgcagtg gctcgcggtg cagaaggagc ttggcctccc gaatgacaag 1020
tcgaacgtca acggcggtgc catcgccctc ggccacccgc tcgcagcgtc cggcgcgcgc 1080
atcaccaaca acctcgtcca ctcgctgcac cggctcaaca agcgcttcgc gatcggctcg 1140
gcgtgcattg gcggcgggca ggcgacgacg atcctctttg agcgggtgta g 1191
<210> 2
<211> 396
<212> PRT
<213> 红冬孢酵母(Rhodosporidium kratochvilovae)
<400> 2
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Gly Gly Ala Leu Lys Asn Tyr Thr Ala Ser Gln Leu Gly Gly Phe Ala
20 25 30
Ala Lys Ala Ala Leu Ala Glu Leu Pro Glu Gly Thr Gln Val Asp Ser
35 40 45
Val Ile Phe Gly Gln Val Leu Tyr Ser Asp Pro Ser Ala Ala Tyr Leu
50 55 60
Ala Arg His Val Gly His His Ala Gly Val Pro Val His Val Pro Ala
65 70 75 80
Leu Thr Val Asn Arg Leu Cys Gly Ser Gly Phe Gln Ala Val Ile Asn
85 90 95
Ala Ala Gln Glu Ile Lys Leu Gly Glu Ser His Val Val Leu Thr Gly
100 105 110
Gly Thr Asp Asn Met Ser Leu Ser Pro Tyr Thr Leu Ser Gly Ser Ser
115 120 125
Arg Phe Gly Asn Lys Tyr Gly Val Asp Leu Lys Leu Glu Asp Ser Leu
130 135 140
Ala Gln Ala Leu Thr Asp Arg Val Pro Asn Pro Thr Pro Met Gly Ile
145 150 155 160
Thr Ala Glu Asn Leu Ala Gln Lys Tyr Gly Ile Ser Arg Glu Gln Cys
165 170 175
Asp Gln Tyr Ala Leu Gln Ser Gln Gln Arg Trp Lys Lys Ala Leu Asp
180 185 190
Ser Gly Ala Phe Glu Ala Glu Ile Thr Pro Val Gln Leu Pro Pro Lys
195 200 205
Lys Arg Gly Gly Glu Pro Ile Thr Phe Ser Gln Asp Glu His Pro Arg
210 215 220
Pro Gln Ala Thr Leu Glu Gln Leu Gly Arg Leu Pro Ala Val Phe Ala
225 230 235 240
Lys Asn Gly Thr Val Thr Ala Gly Asn Ala Ser Gly Ile Cys Asp Gly
245 250 255
Ala Ala Ala Asn Val Val Val Ser Glu Glu Ala Val Lys Arg Tyr Gly
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Leu Lys Pro Leu Ala Arg Val Val Ala Tyr His Ile Asn Ala Val Asp
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Pro Asn Ile Met Gly Ile Gly Pro Val Glu Gly Ile Arg Gly Val Leu
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Lys Lys Ala Gly Met Lys Ile Glu Asp Ile Asp Leu Phe Asp Ile Asn
305 310 315 320
Glu Ala Phe Ala Ala Gln Trp Leu Ala Val Gln Lys Glu Leu Gly Leu
325 330 335
Pro Asn Asp Lys Ser Asn Val Asn Gly Gly Ala Ile Ala Leu Gly His
340 345 350
Pro Leu Ala Ala Ser Gly Ala Arg Ile Thr Asn Asn Leu Val His Ser
355 360 365
Leu His Arg Leu Asn Lys Arg Phe Ala Ile Gly Ser Ala Cys Ile Gly
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Gly Gly Gln Ala Thr Thr Ile Leu Phe Glu Arg Val
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<210> 3
<211> 23
<212> DNA
<213> 人工序列(Artificial)
<400> 3
tcaccatggc cgtcttcatc gcg 23
<210> 4
<211> 27
<212> DNA
<213> 人工序列(Artificial)
<400> 4
cttgatatcc tacacccgct caaagag 27
Claims (2)
1.一种乙酰辅酶A乙酰转移酶基因RKAcaT2,其核苷酸序列如SEQ ID NO:1所示,该基因编码的氨基酸序列如SEQ ID NO:2所示。
2.权利要求1所述的乙酰辅酶A乙酰转移酶基因RKAcaT2在产类胡萝卜素中的应用。
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CN108624600A (zh) * | 2018-05-22 | 2018-10-09 | 昆明理工大学 | 锌指转录因子基因RkMsn4的用途 |
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Citations (25)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2005003721A2 (en) * | 2003-06-09 | 2005-01-13 | Protometrix, Inc. | Enzyme activity assays on protein microarrays |
CN1630718A (zh) * | 2001-06-06 | 2005-06-22 | Dsmip资产公司 | 改进的类异戊二烯生产 |
CN101096651A (zh) * | 2007-06-05 | 2008-01-02 | 清华大学 | 表达聚羟基脂肪酸酯的工程菌及其构建方法与应用 |
CN101218352A (zh) * | 2005-03-18 | 2008-07-09 | 米克罗比亚精确工程公司 | 产油酵母和真菌中类胡萝卜素的产生 |
CN101748069A (zh) * | 2008-12-12 | 2010-06-23 | 中国科学院微生物研究所 | 一种重组蓝藻 |
WO2013066848A1 (en) * | 2011-10-31 | 2013-05-10 | Ginkgo Bioworks, Inc. | Methods and systems for chemoautotrophic production of organic compounds |
CN103361274A (zh) * | 2012-04-05 | 2013-10-23 | 浙江齐成碳能科技有限公司 | 生产乙醇的基因工程微生物 |
CN103974973A (zh) * | 2011-11-09 | 2014-08-06 | 莫茨制药有限及两合公司 | 显示出缩短的生物学活性的神经毒素 |
CN104164457A (zh) * | 2014-07-15 | 2014-11-26 | 青岛农业大学 | 一种利用新mva途径合成异戊二烯的方法及其相应重组细胞和应用 |
CN104471068A (zh) * | 2011-12-16 | 2015-03-25 | 布拉斯科南美公司(巴西) | 经修饰的微生物和使用其制造丁二烯的方法 |
CN104769119A (zh) * | 2012-06-15 | 2015-07-08 | 英威达技术有限责任公司 | 生物合成1,3丁二烯的方法 |
CN104903441A (zh) * | 2012-11-09 | 2015-09-09 | 拉勒曼德匈牙利流动管理有限责任公司 | 在纤维素原料的乙醇发酵期间消耗乙酸盐的方法 |
CN105339490A (zh) * | 2012-12-20 | 2016-02-17 | 帝斯曼知识产权资产管理有限公司 | 乙酰转移酶及其用于产生类胡萝卜素的用途 |
CN107058145A (zh) * | 2009-11-23 | 2017-08-18 | 纽塞利斯公司 | 使用酵母产生角鲨烯的方法和组合物 |
CN107190004A (zh) * | 2016-03-14 | 2017-09-22 | 辛珉 | 一种tigr调控的甲羟戊酸途径 |
CN107794254A (zh) * | 2017-09-30 | 2018-03-13 | 昆明理工大学 | 重组高温裂解酶的快速纯化方法 |
CN108350469A (zh) * | 2015-11-17 | 2018-07-31 | 环球生物能源公司 | 从3-甲基巴豆酸生产异丁烯的方法 |
CN108753802A (zh) * | 2018-05-22 | 2018-11-06 | 昆明理工大学 | 一个苹果酸脱氢酶基因cimdh1及其重组表达载体 |
CN109266596A (zh) * | 2018-09-28 | 2019-01-25 | 中国科学院微生物研究所 | 高效利用脂肪酸合成甘氨酸的大肠杆菌重组菌及其构建方法与应用 |
CN109777815A (zh) * | 2019-03-28 | 2019-05-21 | 昆明理工大学 | HMG-CoA合成酶基因RKHMGCS及其应用 |
KR20190097448A (ko) * | 2018-02-12 | 2019-08-21 | 대한민국(산림청 국립산림과학원장) | 겨울 우산 버섯 유래 테르펜 합성효소 유전자 및 상기 유전자가 삽입된 돌연변이 효모의 형질전환체 |
CN110747206A (zh) * | 2019-11-05 | 2020-02-04 | 昆明理工大学 | 3-羟基-3-甲基戊二酸单酰辅酶a还原酶基因rkhmgr及其应用 |
JP6643083B2 (ja) * | 2012-06-01 | 2020-02-12 | ランザテク・ニュージーランド・リミテッド | 組み換え代謝微生物およびその使用 |
US10750711B2 (en) * | 2015-06-08 | 2020-08-25 | Indigo Ag, Inc. | Streptomyces endophyte compositions and methods for improved agronomic traits in plants |
CN112143747A (zh) * | 2020-09-09 | 2020-12-29 | 昆明理工大学 | 一种噬菌体裂解酶及其基因、基因重组表达载体与应用 |
-
2019
- 2019-02-27 CN CN201910147204.0A patent/CN109666683B/zh active Active
Patent Citations (26)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1630718A (zh) * | 2001-06-06 | 2005-06-22 | Dsmip资产公司 | 改进的类异戊二烯生产 |
WO2005003721A2 (en) * | 2003-06-09 | 2005-01-13 | Protometrix, Inc. | Enzyme activity assays on protein microarrays |
CN101218352A (zh) * | 2005-03-18 | 2008-07-09 | 米克罗比亚精确工程公司 | 产油酵母和真菌中类胡萝卜素的产生 |
CN101096651A (zh) * | 2007-06-05 | 2008-01-02 | 清华大学 | 表达聚羟基脂肪酸酯的工程菌及其构建方法与应用 |
CN101748069A (zh) * | 2008-12-12 | 2010-06-23 | 中国科学院微生物研究所 | 一种重组蓝藻 |
CN107058145A (zh) * | 2009-11-23 | 2017-08-18 | 纽塞利斯公司 | 使用酵母产生角鲨烯的方法和组合物 |
WO2013066848A1 (en) * | 2011-10-31 | 2013-05-10 | Ginkgo Bioworks, Inc. | Methods and systems for chemoautotrophic production of organic compounds |
US10801045B2 (en) * | 2011-10-31 | 2020-10-13 | Ginkgo Bioworks, Inc. | Methods for making chemoautotrophic cells by engineering an energy conversion pathway and a carbon fixation pathway |
CN103974973A (zh) * | 2011-11-09 | 2014-08-06 | 莫茨制药有限及两合公司 | 显示出缩短的生物学活性的神经毒素 |
CN104471068A (zh) * | 2011-12-16 | 2015-03-25 | 布拉斯科南美公司(巴西) | 经修饰的微生物和使用其制造丁二烯的方法 |
CN103361274A (zh) * | 2012-04-05 | 2013-10-23 | 浙江齐成碳能科技有限公司 | 生产乙醇的基因工程微生物 |
JP6643083B2 (ja) * | 2012-06-01 | 2020-02-12 | ランザテク・ニュージーランド・リミテッド | 組み換え代謝微生物およびその使用 |
CN104769119A (zh) * | 2012-06-15 | 2015-07-08 | 英威达技术有限责任公司 | 生物合成1,3丁二烯的方法 |
CN104903441A (zh) * | 2012-11-09 | 2015-09-09 | 拉勒曼德匈牙利流动管理有限责任公司 | 在纤维素原料的乙醇发酵期间消耗乙酸盐的方法 |
CN105339490A (zh) * | 2012-12-20 | 2016-02-17 | 帝斯曼知识产权资产管理有限公司 | 乙酰转移酶及其用于产生类胡萝卜素的用途 |
CN104164457A (zh) * | 2014-07-15 | 2014-11-26 | 青岛农业大学 | 一种利用新mva途径合成异戊二烯的方法及其相应重组细胞和应用 |
US10750711B2 (en) * | 2015-06-08 | 2020-08-25 | Indigo Ag, Inc. | Streptomyces endophyte compositions and methods for improved agronomic traits in plants |
CN108350469A (zh) * | 2015-11-17 | 2018-07-31 | 环球生物能源公司 | 从3-甲基巴豆酸生产异丁烯的方法 |
CN107190004A (zh) * | 2016-03-14 | 2017-09-22 | 辛珉 | 一种tigr调控的甲羟戊酸途径 |
CN107794254A (zh) * | 2017-09-30 | 2018-03-13 | 昆明理工大学 | 重组高温裂解酶的快速纯化方法 |
KR20190097448A (ko) * | 2018-02-12 | 2019-08-21 | 대한민국(산림청 국립산림과학원장) | 겨울 우산 버섯 유래 테르펜 합성효소 유전자 및 상기 유전자가 삽입된 돌연변이 효모의 형질전환체 |
CN108753802A (zh) * | 2018-05-22 | 2018-11-06 | 昆明理工大学 | 一个苹果酸脱氢酶基因cimdh1及其重组表达载体 |
CN109266596A (zh) * | 2018-09-28 | 2019-01-25 | 中国科学院微生物研究所 | 高效利用脂肪酸合成甘氨酸的大肠杆菌重组菌及其构建方法与应用 |
CN109777815A (zh) * | 2019-03-28 | 2019-05-21 | 昆明理工大学 | HMG-CoA合成酶基因RKHMGCS及其应用 |
CN110747206A (zh) * | 2019-11-05 | 2020-02-04 | 昆明理工大学 | 3-羟基-3-甲基戊二酸单酰辅酶a还原酶基因rkhmgr及其应用 |
CN112143747A (zh) * | 2020-09-09 | 2020-12-29 | 昆明理工大学 | 一种噬菌体裂解酶及其基因、基因重组表达载体与应用 |
Non-Patent Citations (7)
Title |
---|
FIRRINCIELI,A.等: ""Rhodotorula graminis WP1 hypothetical protein partial mRNA"", 《GENBANK DATABASE》 * |
NICOLE WERNER等: ""Functional characterization of thiolaseencoding genes from Xanthophyllomyces dendrorhous and their effects on carotenoid synthesis"", 《BMC MICROBIOLOGY》 * |
SANG-HWAL YOON等: ""Combinatorial expression of bacterial whole mevalonate pathway for the production of β-carotene in E. coli"", 《JOURNAL OF BIOTECHNOLOGY》 * |
TKAVC,R.等: ""putative Acetyl-CoA C-acetyltransferase [Rhodotorula taiwanensis]"", 《GENBANK DATABSE》 * |
刘小岭: ""红冬孢酵母类胡萝卜素合成关键酶基因Pdh和Lc的研究"", 《中国优秀硕士学位论文全文数据库基础科学辑》 * |
姚元枝等: ""鱼腥草乙酰辅酶A 酰基转移酶基因克隆、表达及生物信息学分析"", 《中草药》 * |
陶俊等: ""类胡萝卜素合成的相关基因及其基因工程"", 《生物工程学报》 * |
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