CN104619852A - 用改变乌头酸酶基因和/或其调控元件的细菌发酵生产l-赖氨酸的方法 - Google Patents
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Abstract
Description
Claims (14)
- 权利要求1, 发酵生产 L-赖氨酸的方法或者提高 L-赖氨酸的发酵量的方法, 其包括:( 1 ) 改造细菌染色体上乌头酸酶基因和 /或其调控元件, 使其乌头酸酶的酶活性和 / 或表达量降低但不消失; 和,(2) 用步骤 (1 ) 改造而得到的细菌发酵生产 L-赖氨酸。
- 2,改造获得的细菌在发酵生产 L-赖氨酸或者提高 L-赖氨酸的发酵量中的应用,其中, 所述改造获得是改造细菌染色体上乌头酸酶基因和 /或其调控元件而获得, 而且使改造获 得的细菌的乌头酸酶的酶活性和 /或表达量降低但不消失。
- 3, 改造细菌的方法, 其包括改造所述细菌染色体上乌头酸酶基因和 /或其调控元件, 使改造获得的细菌的乌头酸酶的酶活性和 /或表达量降低但不消失。4, 权利要求 1-3之任一所述的方法或应用, 其中, 改造细菌染色体上乌头酸酶基因 是对所述乌头酸酶基因的核苷酸序列进行添加、 缺失或替换一个或多个核苷酸。
- 5, 权利要求 4所述的方法或应用, 其中, 替换包括对所述乌头酸酶基因的起始密码 子进行替换, 优选替换为 GTG。
- 6, 权利要求 4所述的方法或应用, 其中, 缺失包括对所述乌头酸酶基因的核苷酸序 列进行缺失, 优选缺失 1-120个核苷酸, 更优选缺失 1-90个核苷酸, 最优选缺失 90个核 苷酸, 如对所述乌头酸酶基因的核苷酸序列的终止密码子前缺失 90个核苷酸。
- 7, 权利要求 1-6之任一所述的方法或应用, 其中, 所述乌头酸酶基因的核苷酸序列 如 SEQ ID No: 1或 2所示。8, 权利要求 1-3之任一所述的方法或应用, 其中, 改造细菌染色体上乌头酸酶基因 的调控元件是对所述乌头酸酶基因的调控元件的核苷酸序列进行添加、 缺失或替换一个 或多个核苷酸。
- 9, 权利要求 8所述的方法或应用, 其中, 所述调控元件是启动子, 优选所述启动子 的核苷酸序列如 SEQ ID No: 4或 6所示。
- 10, 权利要求 8所述的方法或应用, 其中, 替换包括对所述乌头酸酶基因的启动子 的核苷酸序列进行替换, 优选替换为如 SEQ ID No: 3或 5所示的核苷酸序列。
- 11, 权利要求 8所述的方法或应用, 其中, 所述调控元件是转录阻遏蛋白, 优选所 述转录阻遏蛋白的核苷酸序列如 SEQ ID No: 7所示。12, 权利要求 8所述的方法或应用, 其中, 添加包括对所述乌头酸酶基因的转录阻 遏蛋白的核苷酸序列进行添加, 优选添加串联如 SEQ ID No: 8和 7所示的核苷酸序列。 13, 权利要求 1-12之任一所述的方法或应用, 其中, 所述细菌是埃希氏菌属或棒杆 菌属细菌, 优选是大肠杆菌、 谷氨酸棒杆菌或北京棒杆菌。
- 14, 权利要求 3-13之任一所述的方法改造而得到的细菌。
- 15, 多核苷酸, 其核苷酸序列选自,( a)对如 SEQ ID No: 1所示的核苷酸序列的起始密码子进行替换 (优选替换为 GTG) 后获得的核苷酸序列;(b ) 对如 SEQ ID No: 1或 2所示的核苷酸序列进行缺失 (优选缺失 1-120个核苷 酸, 更优选缺失 1-90个核苷酸, 最优选缺失 90个核苷酸) 后获得的核苷酸序列, 如对 如 SEQ ID No : 1或 2所示的核苷酸序列的终止密码子前缺失 90个核苷酸后获得的核苷 酸序列; 和,( c) 串联如 SEQ ID No: 8和 7所示的核苷酸序列。
- 16, 载体, 其包含权利要求 15所述的多核苷酸。
- 17,权利要求 15所述的多核苷酸和 /或权利要求 16所述的载体在权利要求 1-13之任 一所述的方法或应用中的应用。
- 18, 权利要求 15所述的多核苷酸和 /或权利要求 16所述的载体在制备权利要求 14 所述的细菌中的应用。
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CN201480002118.6A CN104619852B (zh) | 2013-02-08 | 2014-01-07 | 用改变乌头酸酶基因和/或其调控元件的细菌发酵生产l-赖氨酸的方法 |
PCT/CN2014/070228 WO2014121669A1 (zh) | 2013-02-08 | 2014-01-07 | 用改变乌头酸酶基因和/或其调控元件的细菌发酵生产l-赖氨酸的方法 |
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CN109929888A (zh) * | 2017-12-15 | 2019-06-25 | 宁夏伊品生物科技股份有限公司 | 改进的l-赖氨酸的发酵生产方法 |
CN110527637A (zh) * | 2019-07-18 | 2019-12-03 | 中国科学院青岛生物能源与过程研究所 | 一种产乌头酸的土曲霉菌株及其构建方法与应用 |
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KR102127181B1 (ko) * | 2013-02-08 | 2020-06-30 | 닝샤 에펜 바이오테크 코 엘티디 | 아코니트산수화 효소 유전자와(또는) 그 조절요소를 개조한 세균 발효를 통한 l-리신 생산방법 |
CN103146772B (zh) * | 2013-02-08 | 2014-06-18 | 宁夏伊品生物科技股份有限公司 | 用乌头酸酶表达弱化和/或酶活性降低的细菌发酵生产l-赖氨酸的方法 |
CN103695364B (zh) * | 2014-01-14 | 2018-02-02 | 中国科学院天津工业生物技术研究所 | 通过弱化5‑氨基乙酰丙酸脱水酶活性获得5‑氨基乙酰丙酸高产菌株及其应用 |
CN106167772B (zh) * | 2016-06-21 | 2019-05-17 | 中国科学院过程工程研究所 | 一种高产丙酮酸的大肠杆菌基因工程菌及其构建方法和应用 |
CN112080534B (zh) * | 2020-08-14 | 2022-03-22 | 廊坊梅花生物技术开发有限公司 | 高产l-氨基酸的工程菌及其构建方法与应用 |
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CN103131738A (zh) * | 2013-02-08 | 2013-06-05 | 宁夏伊品生物科技股份有限公司 | 用改变乌头酸酶调控元件的细菌发酵生产l-赖氨酸的方法 |
CN103146772A (zh) * | 2013-02-08 | 2013-06-12 | 宁夏伊品生物科技股份有限公司 | 用乌头酸酶表达弱化和/或酶活性降低的细菌发酵生产l-赖氨酸的方法 |
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CN110527637A (zh) * | 2019-07-18 | 2019-12-03 | 中国科学院青岛生物能源与过程研究所 | 一种产乌头酸的土曲霉菌株及其构建方法与应用 |
CN110527637B (zh) * | 2019-07-18 | 2021-10-15 | 中国科学院青岛生物能源与过程研究所 | 一种产乌头酸的土曲霉菌株及其构建方法与应用 |
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