CN116640751A - 天冬氨酸铵离子裂合酶突变体及其应用 - Google Patents
天冬氨酸铵离子裂合酶突变体及其应用 Download PDFInfo
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- CN116640751A CN116640751A CN202210141287.4A CN202210141287A CN116640751A CN 116640751 A CN116640751 A CN 116640751A CN 202210141287 A CN202210141287 A CN 202210141287A CN 116640751 A CN116640751 A CN 116640751A
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- aspartic acid
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Abstract
本发明涉及微生物技术领域,具体涉及天冬氨酸铵离子裂合酶突变体及其应用。本发明提供的天冬氨酸铵离子裂合酶突变体具有如SEQ ID NO.1或2所示的氨基酸序列。该突变体显著增强了天冬氨酸铵离子裂合酶的酶活性,具有明显更高的催化效率,可促进天冬氨酸的合成,进而增强天冬氨酸下游代谢产物的合成和积累。利用该天冬氨酸铵离子裂合酶突变体构建的重组微生物的赖氨酸合成能力显著提升,赖氨酸的产量和转化率得到明显提高。
Description
技术领域
本发明涉及微生物技术领域,具体涉及天冬氨酸铵离子裂合酶突变体及其应用。
背景技术
大肠杆菌(Escherichia coli)是革兰氏阴性细菌,具有生长速度快、遗传背景清晰、代谢工程手段成熟的优点。鉴于此,大肠杆菌广泛应用于工业发酵领域,可用于生产L-氨基酸、核苷酸及其他有机酸等。
赖氨酸是世界上第二大氨基酸生产品种,广泛应用于动物饲料、医药和食品工业。其中约90%的赖氨酸用于饲料工业,10%用于食品和医药行业。赖氨酸在用于动物饲料添加剂时,可帮助机体吸收其他氨基酸从而提高饲料质量。
大肠杆菌中赖氨酸的合成从天冬氨酸开始,经过多步酶催化反应得到赖氨酸。天冬氨酸作为赖氨酸合成的直接前体,其供应的强化可以有效提高赖氨酸的产率。肠杆菌中催化天冬氨酸生成的一个关键酶是天冬氨酸铵离子裂合酶,该酶由aspA基因编码,该酶可将TCA循环的中间产物延胡索酸一步氨化为天冬氨酸。目前,已有通过引入多拷贝基因或启动子强化表达的方式提高天冬氨酸铵离子裂合酶的表达水平的报道,也有文献报道该酶的特定位置的氨基酸的突变可导致酶活性的丧失或下调,但未见有通过突变获得酶活性增强的天冬氨酸铵离子裂合酶突变体的报道。
发明内容
本发明的目的是提供一种天冬氨酸铵离子裂合酶突变体及其应用。本发明的另一目的是提供表达该天冬氨酸铵离子裂合酶突变体的重组微生物及其应用。
具体地,本发明提供以下技术方案:
本发明提供一种天冬氨酸铵离子裂合酶突变体,所述天冬氨酸铵离子裂合酶突变体具有如SEQ ID NO.1或2所示的氨基酸序列。
序列如SEQ ID NO.1和SEQ ID NO.2所示的天冬氨酸铵离子裂合酶突变体分别为在大肠杆菌野生型天冬氨酸铵离子裂合酶的基础上,将其第69位氨基酸突变为天冬氨酸和亮氨酸得到。
本发明发现,将天冬氨酸铵离子裂合酶的第69位氨基酸突变为天冬氨酸和亮氨酸,能够显著提高天冬氨酸铵离子裂合酶的催化活性,天冬氨酸铵离子裂合酶活性的提高可增强天冬氨酸的合成,进而有利于提高天冬氨酸以及以天冬氨酸为前体的代谢产物的合成,其中,将第69位氨基酸突变为天冬氨酸的效果明显更优。
本领域技术人员应该理解,在上述天冬氨酸铵离子裂合酶突变体序列的N端或C端添加标签蛋白或将其与其他蛋白进行融合形成融合蛋白,在不改变上述天冬氨酸铵离子裂合酶本身结构的情况下,其活性不会发生明显改变,因此,上述添加标签的蛋白或融合蛋白也在本发明的保护范围内。
本发明还提供编码以上所述的天冬氨酸铵离子裂合酶突变体的核酸分子。
根据天冬氨酸铵离子裂合酶突变体的氨基酸序列,本领域技术人员能够确定编码该突变体的核酸分子的核苷酸序列。基于密码子的简并性,上述核酸分子的序列不止一种,所有能够编码上述天冬氨酸铵离子裂合酶突变体的核酸分子均在本发明的保护范围内。
本发明还提供含有所述核酸分子的生物材料,所述生物材料为表达盒、载体或宿主细胞。
其中,所述表达盒可为在所述基因的上游或下游连接用于驱动其转录、翻译的元件得到的重组DNA分子。
所述载体可为表达载体或克隆载体,包括但不限于质粒载体、噬菌体载体、病毒载体、转座子等。
所述宿主细胞可为微生物细胞。
在上述天冬氨酸铵离子裂合酶突变体的基础上,本发明提供一种重组微生物,所述重组微生物表达以上所述的天冬氨酸铵离子裂合酶突变体。
以上所述的表达天冬氨酸铵离子裂合酶突变体可采用如下任意一种或多种方式实现:
(1)将出发菌株的染色体上原始的天冬氨酸铵离子裂合酶突变体编码基因替换(突变)为所述天冬氨酸铵离子裂合酶突变体的编码基因;
(2)在出发菌株中导入含有所述天冬氨酸铵离子裂合酶突变体的编码基因的质粒等载体;
(3)在出发菌株的染色体上增加至少1个天冬氨酸铵离子裂合酶突变体编码基因的拷贝。
优选地,所述重组微生物表达以上所述的天冬氨酸铵离子裂合酶突变体,但不表达其出发菌株中原始的天冬氨酸铵离子裂合酶。
进一步优选地,所述重组微生物中,编码天冬氨酸铵离子裂合酶的基因被替换为以上所述的编码所述天冬氨酸铵离子裂合酶突变体的核酸分子。
以上所述的重组微生物为埃希氏菌属细菌,优选为大肠杆菌(Escherichiacoli)。
以上所述的出发菌株是指用于将编码天冬氨酸铵离子裂合酶的基因替换为编码所述天冬氨酸铵离子裂合酶突变体的基因的起始菌株,即:将出发菌株的编码天冬氨酸铵离子裂合酶的基因替换为编码所述天冬氨酸铵离子裂合酶突变体的基因即可得到所述重组微生物。
通过将出发菌株中的天冬氨酸铵离子裂合酶编码基因突变为编码所述天冬氨酸铵离子裂合酶突变体的基因得到的重组微生物,其天冬氨酸或以天冬氨酸为前体的代谢产物的产量和转化率较出发菌株显著提高。
优选地,所述出发菌株为能够合成并积累赖氨酸的大肠杆菌。
作为本发明的一种实施方式,所述出发菌株为诱变获得的产赖氨酸大肠杆菌MHZ-0914,该菌株已经于2021年6月1日保藏于中国微生物菌种保藏管理委员会普通微生物中心(简称CGMCC,地址:北京市朝阳区北辰西路1号院3号,中国科学院微生物研究所,邮编100101),保藏编号为CGMCC No.22648,分类命名为大肠埃希氏菌Escherichia coli。
本发明还提供以上所述的重组微生物的构建方法,所述方法包括:将所述重组微生物的出发菌株中编码天冬氨酸铵离子裂合酶的基因替换为编码所述天冬氨酸铵离子裂合酶突变体的基因。
上述基因的替换可采用本领域的常规技术手段实现,例如:采用同源重组的方法(包括但不限于CRISPR重组系统等)将出发菌株染色体上编码天冬氨酸铵离子裂合酶的基因替换为编码所述天冬氨酸铵离子裂合酶突变体的基因。
本发明进一步提供所述天冬氨酸铵离子裂合酶突变体或所述核酸分子或所述生物材料或所述重组微生物的如下任一种应用:
(1)在构建用于生产天冬氨酸或以天冬氨酸为合成前体的代谢产物的微生物中的应用;
(2)在发酵生产天冬氨酸或以天冬氨酸为合成前体的代谢产物中的应用;
(3)在提高微生物的天冬氨酸或以天冬氨酸为合成前体的代谢产物的产量和/或转化率中的应用。
优选地,以上所述的应用中,所述以天冬氨酸为合成前体的代谢产物为赖氨酸。所述微生物为埃希氏菌属细菌,优选为大肠杆菌(Escherichia coli)。
本发明提供一种发酵生产天冬氨酸或以天冬氨酸为合成前体的代谢产物的方法,所述方法包括:培养以上所述的重组微生物得到培养物,将培养物经分离提取得到天冬氨酸或以天冬氨酸为合成前体的代谢产物。
具体地,上述方法包括:将所述重组微生物接种于种子培养基中进行种子培养,得到种子液,将种子液接种于发酵培养基中培养,得到发酵液,将发酵液经分离提取得到天冬氨酸或以天冬氨酸为合成前体的代谢产物。
优选地,所述以天冬氨酸为合成前体的代谢产物为赖氨酸。
优选地,所述发酵培养基包括如下组分:葡萄糖45-55g/L,硫酸铵20-30g/L,酵母膏3-5g/L,磷酸二氢钾1-2g/L,七水硫酸镁0.8-1.2g/L,硫酸亚铁0.02-0.04g/L,硫酸锰0.02-0.04g/L,碳酸钙0-30g/L,pH6.5-7.5。
本发明的有益效果在于:本发明提供的天冬氨酸铵离子裂合酶突变体显著增强了天冬氨酸铵离子裂合酶的酶活性,该突变体具有明显更高的催化效率,可促进天冬氨酸的合成,进而增强天冬氨酸下游代谢产物的合成和积累。利用该天冬氨酸铵离子裂合酶突变体构建的重组微生物的赖氨酸合成能力显著提升,赖氨酸的产量和转化率得到明显提高。
具体实施方式
本发明提供的天冬氨酸铵离子裂合酶突变体的氨基酸序列如SEQ ID NO.1或SEQID NO.2所示。
本发明还提供表达上述天冬氨酸铵离子裂合酶突变体的重组微生物,所述重组微生物优选为重组大肠杆菌。所述重组大肠杆菌为将出发菌株中原始的天冬氨酸铵离子裂合酶编码基因突变为上述天冬氨酸铵离子裂合酶突变体的编码基因得到。
本发明还提供利用上述重组微生物发酵生产赖氨酸的方法,所述方法包括:培养上述重组微生物得到培养物,将培养物经分离提取得到赖氨酸。
以下实施例用于说明本发明,但不用来限制本发明的范围。
以下实施例中未注明具体技术或条件者,按照本领域内的文献所描述的技术或条件,或者按照产品说明书进行。所用试剂或仪器未注明生产厂商者,均为可通过正规渠道商购买得到的常规产品。
以下实施例中涉及的引物名称及序列如表1所示。
表1引物序列信息
引物名称 | 序列(5’-3’) |
pTF-aspA-sgRNA-F | tcctaggtataatactagtgttcaggacttcatcacatggttttagagctagaaatagc |
pTF-aspA-sgRNA-R | actagtattatacctaggactgagctagctgtcaag |
aspA-P69-UF | gttcagaccagtaccgattg |
aspA-P69L-UR | acaaagagctgcaaaccattctcaaaagtgtagcgaatgccat |
aspA-P69L-DF | atggcattcgctacacttttgagaatggtttgcagctctttgt |
aspA-P69-DR | acctgaatgggttgcgaatc |
aspA-P69D-UR | acaaagagctgcaaaccattgacaaaagtgtagcgaatgccat |
aspA-P69D-DF | atggcattcgctacacttttgtcaatggtttgcagctctttgt |
aspA-P69-F1 | gtgggcctgaagagagcaag |
aspA-P69-R1 | gtgaatataaccagcacgag |
实施例1含有突变基因aspA(P69D)的重组菌的构建
本实施例以大肠杆菌MHZ-0914(保藏编号为CGMCC No.22648)为出发菌株,提供表达SEQ ID NO.1所示的天冬氨酸铵离子裂合酶突变体(天冬氨酸铵离子裂合酶的第69位氨基酸突变为天冬氨酸)的重组大肠杆菌。经测序分析,大肠杆菌MHZ-0914的天冬氨酸铵离子裂合酶的氨基酸序列如SEQ ID NO.3所示。
重组大肠杆菌的构建方法如下:
(1)pTargetF-N20-aspA(P69D)质粒及Donor DNA构建
步骤1:使用pTF-aspA-sgRNA-F/pTF-aspA-sgRNA-R为引物,以质粒pTargetT为模板,扩增得到带有N20的pTF线性质粒,使用无缝组装Clone Express试剂盒将此线性质粒于37℃进行组装,随后转化Trans1-T1感受态细胞,获得pTargetF-N20-aspA(P69D),并进行PCR鉴定及测序验证;该质粒用于提供靶向序列。
步骤2:以MG1655基因组为模板,选用aspA-P69-UF/aspA-P69D-UR引物对,扩增得到上游同源臂①,选用aspA-P69D-DF/aspA-P69-DR引物对,扩增得到下游同源臂②,以①、②为模板,选用aspA-P69-UF/aspA-P69-DR引物对,扩增得到Donor DNA。
(2)感受态细胞制备及电转化
步骤1:制备CGMCC No.22648的感受态细胞,将pCas质粒电转入该感受态细胞中(转化方法及感受态制备方法均参照《分子克隆III》)。
步骤2:挑取步骤1得到的单菌落于5mL含卡那霉素和终浓度为10mM的阿拉伯糖的LB培养基中,30℃,200r/min培养至OD650为0.4后制备电转感受态细胞(感受态制备方法参照《分子克隆III》)。
步骤3:将上述(1)中构建得到的pTargetF-N20-aspA(P69D)质粒和Donor DNA同时电转入带有pCas的感受态细胞中(电转条件:2.5kV,200Ω,25μF),涂布于含壮观霉素和卡那霉素的LB平板上,30℃静置培养至单菌落可见。
(3)重组验证
步骤1:使用引物对aspA-P69-F1/aspA-P69-R1对上述(2)的步骤3中得到的单菌落进行菌落PCR验证;
步骤2:将PCR鉴定正确的菌株用引物对aspA-P69-F1/aspA-P69-R1扩增,扩增产物送测序,以验证序列的完整性。
(4)质粒的消除
步骤1:挑取上述(3)中测序验证正确的单菌落接种于5mL含卡那霉素及终浓度为0.5mM的IPTG的LB培养基中,30℃过夜培养后划线于含卡那霉素的LB平板上;
步骤2:挑取步骤1的单菌落对点于含卡那霉素、壮观霉素LB平板和只含卡那霉素的LB平板上,30℃过夜培养,若在含卡那霉素、壮观霉素的LB平板上不能生长,在卡那霉素的LB平板上生长,表明pTargetF-N20质粒已丢失;
步骤3:挑取pTargetF-N20质粒丢失的阳性菌落,接种于无抗LB培养基中,42℃培养8h后划线于LB平板上,37℃过夜培养;
步骤4:挑取步骤3得到的单菌落对点于含卡那霉素LB平板和无抗LB平板上,若在含卡那霉素的LB平板上不能生长,在无抗LB平板上生长,表明pCas质粒丢失,得到的菌株命名为22648-aspA69D。
实施例2含有突变基因aspA(P69L)的重组菌的构建
本实施例以大肠杆菌MHZ-0914(保藏编号为CGMCC No.22648)为出发菌株,提供表达SEQ ID NO.2所示的天冬氨酸铵离子裂合酶突变体(天冬氨酸铵离子裂合酶的第69位氨基酸突变为亮氨酸)的重组大肠杆菌。
重组大肠杆菌的构建方法如下:
(1)pTargetF-N20-aspA(P69L)质粒及Donor DNA构建
步骤1:使用pTF-aspA-sgRNA-F/pTF-aspA-sgRNA-R为引物,以质粒pTargetT为模板(参见Multigene Editing in the Escherichiacoli Genome via the CRISPR-Cas9System,Jiang Y,Chen B,et al.Appl.EnvironMicrobiol,2015),扩增得到带有N20的pTF线性质粒,使用无缝组装Clone Express试剂盒将此线性质粒于37℃进行组装,随后转化Trans1-T1感受态细胞,获得pTargetF-N20-aspA(P69L),并进行PCR鉴定及测序验证;
步骤2:以MG1655基因组为模板,选用aspA-P69-UF/aspA-P69L-UR引物对,扩增得到上游同源臂①,选用aspA-P69L-DF/aspA-P69-DR引物对,扩增得到下游同源臂②,以①、②为模板,选用aspA-P69-UF/aspA-P69-DR引物对,扩增得到Donor DNA。
(2)感受态细胞制备及电转化
步骤1:将pCas质粒(参见Multigene Editing in the Escherichia coliGenomevia the CRISPR-Cas9 System,Jiang Y,Chen B,et al.Appl.Environ.Microbiol,2015)电转入大肠赖氨酸生产菌CGMCC No.22648的感受态细胞中(转化方法及感受态制备方法均参照《分子克隆III》)。
步骤2:挑取步骤1得到的单菌落于5mL含卡那霉素和终浓度为10mM的阿拉伯糖的LB培养基中,30℃,200r/min培养至OD650为0.4后制备电转感受态细胞(感受态制备方法参照《分子克隆III》)。
步骤3:将上述(1)中构建得到的pTargetF-N20-aspA(P69L)质粒和Donor DNA同时电转入带有pCas的感受态细胞中(电转条件:2.5kV,200Ω,25μF),涂布于含壮观霉素和卡那霉素的LB平板上,30℃静置培养至单菌落可见。
(3)重组验证
步骤1:使用引物对aspA-P69-F1/aspA-P69-R1对上述2的步骤(3)得到的单菌落进行菌落PCR验证;
步骤2:将PCR鉴定正确的菌株用引物对aspA-P69-F1/aspA-P69-R1扩增,扩增产物送测序,以验证序列的完整性。
(4)质粒的消除
步骤1:挑取上述(3)中得到的测序验证正确的单菌落接种于5mL含卡那霉素及终浓度为0.5mM的IPTG的LB培养基中,30℃过夜培养后划线于含卡那霉素的LB平板上;
步骤2:挑取步骤1得到的单菌落对点于含卡那霉素、壮观霉素LB平板和只含卡那霉素的LB平板上,30℃过夜培养,若在含卡那霉素、壮观霉素的LB平板上不能生长,在卡那霉素的LB平板上生长,表明pTargetF-N20质粒已丢失;
步骤3:挑取pTargetF-N20质粒丢失的阳性菌落,接种于无抗LB培养基中,42℃培养8h后划线于LB平板上,37℃过夜培养;
步骤4:挑取步骤3得到的单菌落对点于含卡那霉素LB平板和无抗LB平板上,若在含卡那霉素的LB平板上不能生长,在无抗LB平板上生长,表明pCas质粒丢失,得到的菌株命名为22648-aspA69L。
实施例3重组菌的天冬氨酸铵离子裂合酶活性检测
培养大肠杆菌CGMCC No.22648以及实施例1和2获得的重组菌22648-aspA69D和22648-aspA69L,从培养物中分离蛋白,并检测天冬氨酸铵离子裂合酶的活性,具体方法如下:
将生长到对数期的各菌株的培养物接种到50mL LB种子培养基中,接种后的初始OD600值为0.3,然后培养直到在600nm的吸光值达到10。离心收集菌体,用20mM Tris HCl(pH8.0)洗涤2次,悬浮于相同的缓冲液中。超声破碎细胞,离心收集上清,通过Bradford法(Bradford,M.M 1976.Anal.Biochem.72:248-254)定量测定上清液的蛋白含量,并将上清液用作测定天冬氨酸铵离子裂合酶活性的粗蛋白溶液。
在37℃、pH 8.0的条件下测定天冬氨酸铵离子裂合酶的活性。反应混合物由延胡索酸(50mM),NH4Cl-NH4OH(200mM,pH 8.0)缓冲液组成,并加入上述制备的一定量的粗蛋白溶液以开始反应。通过测量还原氨化反应中天冬氨酸生成的量来间接地计算底物的消耗量,天冬氨酸的浓度由高效液相色谱HPLC方法测定。1单位(U)的酶活性定义为,每分钟催化延胡索酸还原产生1μmol天冬氨酸所需的酶量。
平行进行3组实验,以平均值为最终结果。结果如表2所示,重组菌22648-aspA69D和22648-aspA69L中天冬氨酸铵离子裂合酶的活性为出发菌株CGMCC No.22468的2-3倍,其天冬氨酸铵离子裂合酶的活性较出发菌株CGMCC No.22468均显著提升,且重组菌22648-aspA69D中天冬氨酸铵离子裂合酶的活性提升幅度显著高于22648-aspA69L。
表2天冬氨酸铵离子裂合酶的活性测定
实施例4重组菌株生产赖氨酸的性能
将实施例1和2构建的重组菌进行赖氨酸发酵实验,赖氨酸发酵过程中使用的培养基配方如下:
活化培养基:蛋白胨10g/L,NaCl 10g/L,酵母粉5g/L,18g/L琼脂粉,调节pH至7.0。
种子培养基:葡萄糖10g/L,硫酸铵4g/L,酵母膏2.0g/L,磷酸二氢钾3g/L,七水硫酸镁0.4g/L,硫酸亚铁0.01g/L,硫酸锰0.01g/L,pH7.0。
发酵培养基:葡萄糖50g/L,硫酸铵25g/L,酵母膏4.0g/L,磷酸二氢钾1.6g/L,七水硫酸镁1.0g/L,硫酸亚铁0.03g/L,硫酸锰0.03g/L,碳酸钙25,pH7.0。
赖氨酸发酵方法如下:
(1)种子活化:从冻存管中取待验证菌株,在种子活化培养基上划线活化,37℃培养12h;
(2)种子培养:挑取平板活化种子1环接至装有20mL种子培养基的500mL三角瓶中,33℃,220r/min振荡培养7h,得到种子液;
(3)发酵培养:将2mL种子液接种至装有20mL发酵培养基的500mL三角瓶中,33℃、220r/min振荡培养12h,得到发酵液。
(4)取2mL发酵液离心(12000rpm,2min),收集上清液,用HPLC检测重组菌与对照菌发酵液中的L-赖氨酸含量,L-赖氨酸含量检测结果如表3表示(表3中数据为三批次实验的平均值)。
表3重组菌生产赖氨酸的性能测试
以上发酵结果表明,出发菌株CGMCC No.22648的L-赖氨酸积累量为18.5g/L,转化率为37%。重组菌22648-aspA69D相比于出发菌和重组菌22648-aspA69L的赖氨酸产量及转化率均有显著提升。
虽然,上文中已经用一般性说明及具体实施方案对本发明作了详尽的描述,但在本发明基础上,可以对之作一些修改或改进,这对本领域技术人员而言是显而易见的。因此,在不偏离本发明精神的基础上所做的这些修改或改进,均属于本发明要求保护的范围。
序列表
<110> 廊坊梅花生物技术开发有限公司
<120> 天冬氨酸铵离子裂合酶突变体及其应用
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Claims (10)
1.天冬氨酸铵离子裂合酶突变体,其特征在于,所述天冬氨酸铵离子裂合酶突变体具有如SEQ ID NO.1或2所示的氨基酸序列。
2.核酸分子,其特征在于,其编码权利要求1所述的天冬氨酸铵离子裂合酶突变体。
3.含有权利要求2所述的核酸分子的生物材料,其特征在于,所述生物材料为表达盒、载体或宿主细胞。
4.一种重组微生物,其特征在于,所述重组微生物表达权利要求1所述的天冬氨酸铵离子裂合酶突变体。
5.根据权利要求4所述的重组微生物,其特征在于,所述重组微生物中,编码天冬氨酸铵离子裂合酶的基因被替换为权利要求2所述的核酸分子。
6.根据权利要求4或5所述的重组微生物,其特征在于,所述重组微生物为埃希氏菌属细菌,优选为大肠杆菌(Escherichia coli)。
7.权利要求4~6任一项所述的重组微生物的构建方法,其特征在于,包括:将所述重组微生物的出发菌株中编码天冬氨酸铵离子裂合酶的基因替换为编码权利要求1所述的天冬氨酸铵离子裂合酶突变体的基因。
8.权利要求1所述的天冬氨酸铵离子裂合酶突变体或权利要求2所述的核酸分子或权利要求3所述的生物材料或权利要求4~6任一项所述的重组微生物的如下任一种应用:
(1)在构建用于生产天冬氨酸或以天冬氨酸为合成前体的代谢产物的微生物中的应用;
(2)在发酵生产天冬氨酸或以天冬氨酸为合成前体的代谢产物中的应用;
(3)在提高微生物的天冬氨酸或以天冬氨酸为合成前体的代谢产物的产量和/或转化率中的应用。
9.根据权利要求8所述的应用,其特征在于,所述以天冬氨酸为合成前体的代谢产物为赖氨酸,和/或,所述微生物为埃希氏菌属细菌,优选为大肠杆菌(Escherichia coli)。
10.一种发酵生产天冬氨酸或以天冬氨酸为合成前体的代谢产物的方法,其特征在于,包括:培养权利要求4~6任一项所述的重组微生物得到培养物,将培养物经分离提取得到天冬氨酸或以天冬氨酸为合成前体的代谢产物;
优选地,所述以天冬氨酸为合成前体的代谢产物为赖氨酸。
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