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1. 发明申请

WO2021036788A1 一种2-酮基-L-古龙酸生产菌株及其构建方法审中-公开
公开(公告)号：WO2021036788A1
公开(公告)日：2021-03-04
申请号：PCT/CN2020/108609
申请日：2020-08-12
申请人：中国科学院分子植物科学卓越创新中心
发明人：杨晟 , 汪庆卓 , 段春兰 , 刘映淼 , 刁刘洋 , 蒋宇 , 陶荣盛
IPC分类号： C12N1/21 , C12P7/60 , C12R1/185
摘要：提供了一种构建2-酮基-L-古龙酸生产菌株的方法，包括任意顺序的如下步骤中的一个以上：1)中断塔特姆氏菌染色体中膜结合葡萄糖脱氢酶编码基因；2)中断葡萄糖激酶编码基因；3)中断葡萄糖酸激酶编码基因；4)中断2-酮醛糖还原酶编码基因和/或其同工酶编码基因；5)重组表达NAD(P)+型葡萄糖脱氢酶；6)重组表达NAD(P)H型2,5-二酮基-D-葡萄糖酸还原酶；7)过表达2,5-二酮基-D-葡萄糖酸内运蛋白ARU92990和ARU92994；8)中断细胞染色体中PTS组分编码基因，得到基因工程菌。该方法构建的2-酮基-L-古龙酸生产菌可以通过一步发酵法或全细胞催化以葡萄糖为原料生产2-酮基-L-古龙酸。

2. 发明申请

WO2005012486A3 METABOLICALLY ENGINEERED BACTERIAL STRAINS HAVING ENHANCED 2-KETO-D-GLUCONATE ACCUMULATION 审中-公开
标题翻译：具有增强的2-羟色胺-D-葡萄糖酸盐积累的代谢工程细菌菌株
公开(公告)号：WO2005012486A3
公开(公告)日：2006-01-12
申请号：PCT/US2004024203
申请日：2004-07-27
申请人： GENENCOR INT , DODGE TIMOTHY C , VALLE FERNANDO , RASHID M HARUNUR
发明人： DODGE TIMOTHY C , VALLE FERNANDO , RASHID M HARUNUR
IPC分类号： C12N20060101 , C12N1/21 , C12N9/04 , C12N15/74 , C12P7/58 , C12P7/60 , C12P19/02 , C12P1/00 , C07H21/04 , C12P1/04 , C12P7/00 , C12P7/26
CPC分类号： C12P7/60 , C12N9/0006 , C12P19/02
摘要： The present invention relates to a method of altering bacterial host cells to accumulate 2-keto-D-gluconic acid (2-KDG) by inactivating an endogenous membrane bound 2-keto-D-gluconate dehydrogenase (2-KDGDH), which prior to inactivation catalyzed the conversion of 2-KDG to 2,5-diketogluconate (2,5-DKG).
摘要翻译：本发明涉及通过灭活内源性膜结合的2-酮-D-葡萄糖酸脱氢酶（2-KDGDH）来改变细菌宿主细胞以积累2-酮-D-葡萄糖酸（2-KDG）的方法，其在失活催化了2-KDG转化为2,5-二酮葡萄糖酸盐（2,5-DKG）。

3. 发明申请

WO2004040955B1 MICROBIOLOGICAL METHOD FOR PRODUCING ASCORBIC ACID 审中-公开
标题翻译：用于生产抗坏血酸的微生物学方法
公开(公告)号：WO2004040955B1
公开(公告)日：2004-09-02
申请号：PCT/US0334465
申请日：2003-10-30
申请人： AVENIR GENETICS LLC
发明人： KADO CLARENCE I , PUJOL CATHERINE J , CHAN ALEXANDER
IPC分类号： C12N1/21 , C12P17/04 , C12P7/60 , C12N1/14 , C12N1/20 , C12N9/04 , C12N15/00
CPC分类号： C12P17/04
摘要： There is provided a microbiological one-step method for the manufacture of ascorbic acid. The method comprises use of the operon expressing 2-ketogluconate dehydrogenase in naturally occurring microorganisms and microorganisms transformed with DNA encoding the 2-ketogluconate dehydrogenase gene. Ascorbic acid is obtained from culturing microorganisms expressing 2-ketogluconate dehydrogenase.
摘要翻译：提供了用于制备抗坏血酸的微生物一步法。该方法包括在天然存在的微生物中使用表达2-酮葡萄糖酸脱氢酶的操纵子和用编码2-酮葡糖酸脱氢酶基因的DNA转化的微生物。通过培养表达2-酮葡萄糖酸脱氢酶的微生物获得抗坏血酸。

4. 发明申请

WO02081631A3 UNCOUPLED PRODUCTIVE AND CATABOLIC HOST CELL PATHWAYS 审中-公开
标题翻译：未经协调的生产性和可驱动的宿主细胞途径
公开(公告)号：WO02081631A3
公开(公告)日：2002-11-28
申请号：PCT/US0210575
申请日：2002-04-04
申请人： GENENCOR INT , DODGE TIMOTHY C , VALLE FERNANDO
发明人： DODGE TIMOTHY C , VALLE FERNANDO
IPC分类号： C12N15/09 , C12N1/21 , C12N9/12 , C12N15/52 , C12P7/60 , C12P19/02 , C12P19/30 , C12P25/00 , C12R1/01 , C12N9/00 , C07H21/04 , C12N1/20
CPC分类号： C12N9/1205 , C12N15/52 , C12P7/60 , C12P19/02 , C12P25/00
摘要： The invention provides methods for producing products comprising improved host cells genetically engineered to have uncoupled productive and catabolic pathways. In particular, the present invention provides host cells having a modification in nucleic acid encoding an endogenous enzymatic activity that phosphorylates D-glucose at its 6th carbon and/or a modification of nucleic acid encoding an enzymatic activity that phosphorylates D-gluconate at its 6th carbon. Such improved host cells are used for the production of products, such as, ascorbic acid intermediates. Methods for making and using the improved host cells are provided. Nucleic acid and amino acid sequences for glucokinase and gluconokinase are provided.
摘要翻译：本发明提供了用于生产产品的方法，所述产品包含基因工程化以具有未偶联的生产和分解代谢途径的改良的宿主细胞。具体而言，本发明提供了在编码内源酶活性的核酸中具有修饰的宿主细胞，所述内源酶活性在其第6个碳处磷酸化D-葡萄糖和/或编码在其第6个碳处磷酸化D-葡萄糖酸的酶活性的核酸的修饰。这种改进的宿主细胞用于生产产品，例如抗坏血酸中间体。提供了制备和使用改进的宿主细胞的方法。提供了葡萄糖激酶和葡萄糖激酶的核酸和氨基酸序列。

5. 发明申请

WO9961647A9 ELECTRODIALYSIS METHODS FOR PURIFICATION AND RECOVERY OF GLUCONIC ACID DERIVATIVES 审中-公开
标题翻译：葡萄糖酸衍生物的纯化和回收的电化学方法
公开(公告)号：WO9961647A9
公开(公告)日：2001-03-08
申请号：PCT/US9911671
申请日：1999-05-26
申请人： ELECTROSYNTHESIS CO INC
发明人： GENDERS J DAVID , GOPAL RAM , HARTSOUGH DAN M , KENDALL PETER M , LONG WILLIAM J , MAZUR DUANE J , ZAPPI GUILLERMO D
IPC分类号： B01D15/04 , B01D61/44 , C02F1/469 , C07H7/00 , C12P7/60
CPC分类号： C12P7/60 , B01D61/44 , B01D61/445 , C07H7/00 , Y02P20/142
摘要： A process for purifying and concentrating a gluconic acid derivative, such as 2-keto-L-gulonic acid, comprising introducing a non-viable and/or acidified fermentation medium or an in-vitro reactor medium comprising at least the gluconic acid derivative and/or salt thereof to electrodialysis thereby purifying and concentrating the gluconic acid derivative.
摘要翻译：一种用于纯化和浓缩葡糖酸衍生物如2-酮-L-古洛糖酸的方法，包括将不存在和/或酸化的发酵培养基或体外反应器培养基导入至少包含葡萄糖酸衍生物和/ 或其盐进行电渗析，从而纯化和浓缩葡萄糖酸衍生物。

6. 发明申请

WO99020772A1 PLASMID VECTORS 审中-公开
公开(公告)号：WO99020772A1
公开(公告)日：1999-04-29
申请号：PCT/JP1998/004611
申请日：1998-10-13
IPC分类号： C12N1/21 , C12N9/04 , C12N15/74 , C12P7/60 , C12N15/63 , C12P21/02
CPC分类号： C12N9/0006 , C12N15/74 , C12P7/60
摘要： DNAs originating in a plasmid pF4 containing the domain controlling the autonomous replication in a bacterium belonging to the genus Gluconobacter and containing a domain from which polynucleotides in the region unnecessary in the autonomous replication have been wholly or partly deleted; in particular, the above DNAs containing as the domain necessary in the autonomous replication the whole region consisting of the bases at the 2897- to 3969-positions in the sequence represented by SEQ ID NO:1 in the Sequence Listing or a part of the same; plasmid vectors containing these DNAs; transformants produced by these plasmid vectors; and a process for producing physiologically active substances by culturing these transformants. By specifying the whole base sequence of the plasmid pF4 and the domain involving the DNA relating to the function of controlling the autonomous replication in a bacterium belonging to the genus Gluconobacter, domains unnecessary in the autonomous replication can be deleted and thus a lessened pF4 can be constructed. By lessening the domain for the autonomous replication in a vector, the other domains of the vector can be enlarged, which makes it possible to integrate a greater variety of structural genes thereinto and to impart more functions thereto.
摘要翻译：源自质粒pF4的DNA，其含有控制属于葡糖杆菌属的细菌的自主复制的结构域，并且含有在自主复制中不需要的区域中的多核苷酸已被全部或部分缺失的结构域; 特别地，上述含有自主复制所必需的结构域的DNA由序列表中SEQ ID NO：1所示序列中2897位至3969位的碱基组成， ; 含有这些DNA的质粒载体; 由这些质粒载体产生的转化体; 以及通过培养这些转化体来生产生理活性物质的方法。通过指定质粒pF4的整个碱基序列和涉及与在属于葡糖杆菌属的细菌中控制自主复制的功能相关的DNA的结构域，可以删除自主复制中不必要的结构域，因此减少的pF4可以是建。通过减少载体中自主复制的结构域，可以扩增载体的其他结构域，这使得可以将更多种结构基因整合到其中并赋予其更多功能。

7. 发明申请

WO99020763A1 D-SORBITOL DEHYDROGENASE, GENES THEREOF AND USE OF THE SAME 审中-公开
标题翻译： D-SORBITOL DEHYDROGENASE，其基因及其用途
公开(公告)号：WO99020763A1
公开(公告)日：1999-04-29
申请号：PCT/JP1998/004612
申请日：1998-10-13
IPC分类号： C12N1/21 , C12N9/04 , C12N15/13 , C12P7/60 , C12P19/02
CPC分类号： C12N9/0006 , C12Y101/99021
摘要： Three subunits constituting D-sorbitol dehydrogenase (SLDH); genes encoding them; a process for producing SLDH by culturing cells transformed by expression vectors containing promoter genes appropriate for expressing the above genes and the above-mentioned genes; and a process for producing L-sorbose or 2-keto-L-gulonic acid (2KLGA) by using the above culture. This process permits 2KLGA as a precursor of L-ascorbic acid to be easily produced in a large amount.
摘要翻译：构成D-山梨醇脱氢酶（SLDH）的三个亚基; 编码它们的基因通过培养由含有适于表达上述基因的启动子基因和上述基因的表达载体转化的细胞产生SLDH的方法; 和通过使用上述培养物生产L-山梨糖或2-酮-L-古洛糖酸（2KLGA）的方法。该方法允许2KLGA作为L-抗坏血酸的前体容易地大量生产。

8. 发明申请

WO98059054A1 IMPROVED ENTEROBACTERIACEAE FERMENTATION STRAINS 审中-公开
标题翻译：改良肠内营养发酵菌株
公开(公告)号：WO98059054A1
公开(公告)日：1998-12-30
申请号：PCT/US1998/012945
申请日：1998-06-22
IPC分类号： C07K14/24 , C12N15/31 , C12N15/74 , C12P7/60 , C12N1/20 , C12N1/21 , C12N15/67
CPC分类号： C07K14/24 , C12N15/74 , C12P7/60
摘要： Methods are provided for preparing improved fermentation strains of the family Enterobacteriaceae which comprise the steps of eliminating the cryptic plasmid from the progenitor strain thereby creating the improved strain. Methods for reducing the mobilization properties of resident plasmids in an Enterobacteriaceae strain containing a cryptic plasmid are also provided. The present invention provides the nucleic acid sequence of pS, a cryptic plasmid found in Pantoea which can be used to identify the cryptic plasmid in strains of Enterobacteriaceae.
摘要翻译：提供了用于制备肠杆菌科的改良发酵菌株的方法，其包括从祖细胞菌株中消除神经质粒的步骤，从而产生改良的菌株。还提供了用于降低含有隐性质粒的肠杆菌科菌株中驻留质粒的动员特性的方法。本发明提供了在Pantoea中发现的可用于鉴定肠杆菌科菌株中的隐性质粒的隐神质的pS的核酸序列。

9. 发明申请

WO2008144792A4 METHOD FOR PRODUCING ASCORBIC ACID USING PECTOBACTER CYPRIPEDII 审中-公开
标题翻译：用于生产抗坏血酸的黑腐cypripedii
公开(公告)号：WO2008144792A4
公开(公告)日：2009-03-05
申请号：PCT/AT2008000185
申请日：2008-05-29
申请人： VOGELBUSCH GMBH , PACHER CLAUDIA , KULBE KLAUS D , STEINER EVA , REMBART GUENTER
发明人： PACHER CLAUDIA , KULBE KLAUS D , STEINER EVA , REMBART GUENTER
IPC分类号： C12P17/04 , C12P7/60
CPC分类号： C12P17/04 , C12P7/60
摘要： The invention relates to a method for producing ascorbic acid, according to which in a first step a carbon source is converted by fermentation into 2,5-diketo-D-gluconic acid, which is then enzymatically reacted with 2,5-diketo-D-gluconic acid reductase from Corynebacterium glutamicum to produced 2-keto-L-gulonic acid. To produce higher product yields, the carbon source is glucose and the conversion from glucose into 2,5-diketo-D-gluconic acid is directly produced by the Pectobacter cypripedii strain HEPO1 (DSMZ 12393).
摘要翻译：本发明涉及一种用于制造抗坏血酸，其中，在第一步骤中，碳源是通过发酵在2,5-二酮-D-葡糖酸转化，然后将其在第二步骤中酶促与2,5-二酮-D-葡糖酸的方法从谷氨酸棒杆菌还原酶转化为2-酮基-L-古洛糖酸。对于更高的产物产率的目的，所述碳源是葡萄糖直接和葡萄糖的至2,5-二酮-D-葡糖酸是由Pectobacter cypripedii HEPOl应变进行转换（DSMZ 12393）。

10. 发明申请

WO2004029262A3 PRODUCTION OF 2 - KETO - L - GULONIC ACD 审中-公开
标题翻译：生产2 - KETO - L - GULONIC ACD
公开(公告)号：WO2004029262A3
公开(公告)日：2004-07-01
申请号：PCT/EP0310490
申请日：2003-09-22
申请人： DSM IP ASSETS BV , HOSHINO TATSUO , SUGISAWA TERUHIDE , TAKAGI YOSHINORI
发明人： HOSHINO TATSUO , SUGISAWA TERUHIDE , TAKAGI YOSHINORI
IPC分类号： C12P7/60
CPC分类号： C12P7/60
摘要： The present invention relates to a process for the production of 2-KGA or a salt thereof from D-sorbitol by a multi-stage continuous or a single-stage semi-continuous fermentation mode using a mixed culture of microorganisms of the genus Gluconobacter or Acetobacter. One of the microorganisms is G. oxydans DSM 4025.
摘要翻译：本发明涉及使用葡萄糖酸杆菌属或醋酸杆菌属的微生物的混合培养的多阶段连续或单阶段半连续发酵模式从D-山梨糖醇生产2-KGA或其盐的方法。其中一种微生物是G.oxydans DSM 4025。

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