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1. 发明申请

WO2006133898A3 IMPROVED BIOSYNTETIC PRODUCTION OF 2, 3-TRANS-CHD 审中-公开
标题翻译：改进2，3-TRANS-CHD的生产生产
公开(公告)号：WO2006133898A3
公开(公告)日：2007-10-04
申请号：PCT/EP2006005664
申请日：2006-06-13
申请人： DSM IP ASSETS BV , DSM BIOTECH GMBH , BONGAERTS JOHANNES JOSEF , RAEVEN LEON JEAN RENIER MARIE , SPRENGER GEORG , KUHM ANDREA , TAKORS RALF , BUJNICKI ROBERT PIOTR
发明人： BONGAERTS JOHANNES JOSEF , RAEVEN LEON JEAN RENIER MARIE , SPRENGER GEORG , KUHM ANDREA , TAKORS RALF , BUJNICKI ROBERT PIOTR
IPC分类号： C12P7/42
CPC分类号： C12P7/42
摘要： The invention relates to a process for the biosynthetic production of [5S,6S]-5,6-dihydroxy-cyclohexa-1,3-diene-1-carboxylic acid (2,3-trans-CHD) in an Escherichia microorganism having increased activity of isochorismate synthase and isochorismatase, and being deficient in activity of 2,3-dihydroxybenzoate synthase, wherein the process is carried out in a chorismate overproducing host microorganism also having decreased glucose dehydrogenase activity, or is carried out in a chorismate overproducing host microorganism naturally lacking glucose dehydrogenase activity. In this improved biosynthetic process, 2,3-trans-CHD is being produced in high yields and at concentrations in the biosynthetic reaction medium of at least 12 g/l. The invention further relates to the use of the supernatant of a process for the biosynthetic production of [5S,6S]-5,6-dihydroxy-cyclohexa-1,3-diene-1-carboxylic acid (2,3-trans-CHD) where the concentration of 2,3-trans-CHD in the process has arrived at a concentration of at least 12 g/l as a source for a co-factor for PQQ dependent enzymes.
摘要翻译：本发明涉及在具有增加的大肠杆菌中生物合成生产[5S，6S] -5,6-二羟基 - 环己-1,3-二烯-1-羧酸（2,3-反式-CHD）的方法等位基因合成酶和异激酶活性，并且缺乏2,3-二羟基苯甲酸合成酶的活性，其中该方法在也具有降低的葡萄糖脱氢酶活性的过度生产的宿主微生物中进行，或者在过量产生宿主微生物的天竺葵中进行缺乏葡萄糖脱氢酶活性。在这种改进的生物合成过程中，以高产率和生物合成反应介质中的浓度产生至少12g / l的2,3-反式CHD。本发明还涉及[5S，6S] -5,6-二羟基 - 环己-1,3-二烯-1-羧酸（2,3-反式-CHD）生物合成生产方法的上清液的用途），其中该过程中2,3-反式CHD的浓度已达到至少12g / l的浓度作为PQQ依赖性酶辅因子的来源。

2. 发明申请

WO2005118829A2 BIOSYNTHETIC PRODUCTION OF 4-AMINO-4-DEOXYCHORISMATE (ADC) AND [3R,4R]-4-AMINO-3-HYDROXYCYCLOHEXA-1,5-DIENE-1-CARBOXYLIC ACID (3,4-CHA) 审中-公开
标题翻译： 4-氨基-4-脱氧木糖（ADC）和[3R，4R] -4-氨基-3-羟基环己基-1,5-二苯基-1-羧酸（3,4-CHA）的生产生产
公开(公告)号：WO2005118829A2
公开(公告)日：2005-12-15
申请号：PCT/EP2005/005988
申请日：2005-06-02
申请人： DSM IP ASSETS B.V. , DSM BIOTECH GMBH , FORSCHUNGSZENTRUM JÜLICH GMBH , WUBBOLTS, Marcel, Gerhardus , BOVENBERG, Roelof, Ary, Lans , SPRENGER, Georg , BONGAERTS, Johannes, Josef , KOZAK, Stefan , MÜLLER, Michael
发明人： WUBBOLTS, Marcel, Gerhardus , BOVENBERG, Roelof, Ary, Lans , SPRENGER, Georg , BONGAERTS, Johannes, Josef , KOZAK, Stefan , MÜLLER, Michael
IPC分类号： C12P13/00
CPC分类号： C12P13/04
摘要： The invention relates to a process for the biosynthetic production of 4-amino-4-deoxychorismate (ADC) performed fermentatively in vivo with a 4-amino-4-deoxychorismate synthase, preferably a PabAB bipartite protein (which may be a fusion protein), at an increased level of activity, thereby obtaining a broth comprising ADC and 4-amino-4-deoxyprephenate (ADP), that are recovered. The invention also relates to a further process of converting the ADP into p-aminophenylalanine. The invention, moreover relates to biosynthetic production of [3R,4R]-4-amino-3-hydroxycyclohexa-1,5-diene-1-carboxylic acid (3,4-CHA), by concerted action of such 4-amino-4-deoxychorismate synthase and of an enzyme capable of converting isochorismate into [5S,6S]-5,6 dihydroxycyclohexa-1,3-diene-1carboxylic acid (2,3-CHD), preferably a phenazine biosynthesis protein PhzD, including recovery of 3,4-CHA. The invention also relates to expression vectors and host cells for use in any of such processes. The invention further relates to the use of 3,4-CHA as a catalytically active product, in particular as a chiral catalyst. And the invention finally relates to synthesis of oseltamivir phosphate from 3,4-CHA.
摘要翻译：本发明涉及用4-氨基-4-脱氧激酶合酶，优选PabAB二分体蛋白（可以是融合蛋白）在体内发酵生物合成生产4-氨基-4-脱氧胸苷酸（ADC）的方法。在增加的活性水平上，从而获得包含ADC和4-氨基-4-脱氧降钙素（ADP）的肉汤，其被回收。本发明还涉及将ADP转化为对氨基苯丙氨酸的另一种方法。本发明还涉及通过这种4-氨基-3-羟基环己基-1,5-二烯-1-羧酸（3,4-CHA）的共同作用生产合成[3R，4R] -4-氨基-3-羟基环己-1,5-二烯-1-羧酸（3,4-CHA） 4-脱氧胸苷酸合酶和能够将异草酸转化为[5S，6S] -5,6二羟基环己-1,3-二烯-1-羧酸（2,3-CHD），优选吩嗪生物合成蛋白PhzD的酶的酶，包括回收的3,4-CHA。本发明还涉及用于任何这些方法的表达载体和宿主细胞。本发明还涉及3,4-CHA作为催化活性产物，特别是作为手性催化剂的用途。本发明最终涉及从3,4-CHA合成磷酸奥司他韦。

3. 发明申请

WO2003093490A1 VERFAHREN ZUR MIKROBIELLEN HERSTELLUNG VON AROMATISCHEN AMINOSÄUREN UND ANDEREN METABOLITEN DES AROMATISCHEN AMINOSÄUREBIOSYNTHESEWEGES 审中-公开
标题翻译： METHOD FOR芳香族氨基酸和芳香族氨基酸的其它代谢物的微生物生产
公开(公告)号：WO2003093490A1
公开(公告)日：2003-11-13
申请号：PCT/DE2003/001380
申请日：2003-04-29
申请人： FORSCHUNGSZENTRUM JÜLICH GMBH , HOLLAND SWEETENER COMPANY V.O.F. , DSM BIOTECH GMBH , ANDERLEI, Britta , SPRENGER, Georg , SAHM, Hermann , BONGAERTS, Johannes
发明人： ANDERLEI, Britta , SPRENGER, Georg , SAHM, Hermann , BONGAERTS, Johannes
IPC分类号： C12P13/22
CPC分类号： C12P13/22 , C12Y604/01001
摘要： Die Erfindung betrifft ein Verfahren zur mikrobiellen Herstellung von aromatischen Aminosäuren und anderen Metaboliten des aromatischen Aminosäurebiosyntheseweges. Mikrobiell hergestellte Substanzen, wie Feinchemikalien, insbesondere aromatische Aminosäuren oder Metabolite des Aromatenbiosyntheseweges sind von großem wirtschaftlichen Interesse, wobei der Bedarf an z.B. Aminosäuren weiterhin zunimmt. Die Erfinder fanden heraus, daß nach Einbringen einer pyc-Gensequenz in Mikroorganismen, in verbesserter Weise aromatische Aminosäuren sowie Metabolite des aromatischen Biosyntheseweges produziert werden konnten. Das erfindungsgemäße Verfahren eignet sich insbesondere zur Herstellung von L-Phenylalanin.
摘要翻译：本发明涉及一种用于微生物生产的芳族氨基酸和芳族氨基酸的其它代谢物的方法。微生物产生的物质，如精细化学品，特别是芳族氨基酸或Aromatenbiosyntheseweges的代谢物是巨大的经济利益的，与需要例如氨基酸不断提高。本发明人已发现，芳族氨基酸以改进的方式，以及芳族生物合成途径的代谢物可以通过在微生物中引入PYC基因序列来制备。本发明的方法特别适合于生产L-苯丙氨酸的。

4. 发明申请

WO2006133898A2 IMPROVED BIOSYNTHETIC PRODUCTION OF [5S,6S]-5,6-DIHYDROXYCYCLOHEXA-1,3-DIENE-1-CARBOXYLIC ACID (2,3-TRANS-CHD) 审中-公开
标题翻译： [5S，6S] -5,6-二羟基环己基-1,3-二苯基-1-羧酸（2,3-TRANS-CHD）的改进的生产生产方法
公开(公告)号：WO2006133898A2
公开(公告)日：2006-12-21
申请号：PCT/EP2006/005664
申请日：2006-06-13
申请人： DSM IP ASSETS B.V. , DSM BIOTECH GMBH , BONGAERTS, Johannes, Josef , RAEVEN, Leon, Jean, Renier, Marie , SPRENGER, Georg , KUHM, Andrea , TAKORS, Ralf , BUJNICKI, Robert, Piotr
发明人： BONGAERTS, Johannes, Josef , RAEVEN, Leon, Jean, Renier, Marie , SPRENGER, Georg , KUHM, Andrea , TAKORS, Ralf , BUJNICKI, Robert, Piotr
IPC分类号： C12P7/42
CPC分类号： C12P7/42
摘要： The invention relates to a process for the biosynthetic production of [5S,6S]-5,6-dihydroxy-cyclohexa-1 ,3-diene-1-carboxylic acid (2,3- trans -CHD) in an Escherichia microorganism having increased activity of isochorismate synthase and isochorismatase, and being deficient in activity of 2,3-dihydroxybenzoate synthase, wherein the process is carried out in a chorismate overproducing host microorganism also having decreased glucose dehydrogenase activity, or is carried out in a chorismate overproducing host microorganism naturally lacking glucose dehydrogenase activity. In this improved biosynthetic process, 2,3- trans -CHD is being produced in high yields and at concentrations in the biosynthetic reaction medium of at least 12 g/l. The invention further relates to the use of the supernatant of a process for the biosynthetic production of [5S,6S]-5,6-dihydroxy-cyclohexa-1 ,3-diene-1-carboxylic acid (2,3- trans -CHD) where the concentration of 2,3-trans-CHD in the process has arrived at a concentration of at least 12 g/l as a source for a co-factor for PQQ dependent enzymes.
摘要翻译：本发明涉及在具有增加的大肠杆菌中的[5S，6S] -5,6-二羟基 - 环己-1,3-二烯-1-羧酸（2,3-反式-CHD）的生物合成生产方法等位基因合成酶和异激酶活性，并且缺乏2,3-二羟基苯甲酸合成酶的活性，其中该方法在也具有降低的葡萄糖脱氢酶活性的过度生产的宿主微生物中进行，或者在过量产生宿主微生物的天竺葵中进行缺乏葡萄糖脱氢酶活性。在这种改进的生物合成过程中，以高产率和生物合成反应介质中的浓度产生至少12g / l的2,3-反式CHD。本发明还涉及生物合成生产[5S，6S] -5,6-二羟基 - 环己-1,3-二烯-1-羧酸（2,3-反式-CHD）的方法的上清液的用途），其中该过程中2,3-反式CHD的浓度已达到至少12g / l的浓度作为PQQ依赖性酶辅因子的来源。

5. 发明申请

WO2002034921A3 FERMENTATIVE PRODUCTION OF D-p-HYDROXYPHENYLGLYCINE AND D-PHENYLGLYCINE 审中-公开
公开(公告)号：WO2002034921A3
公开(公告)日：2002-05-02
申请号：PCT/NL2001/000772
申请日：2001-10-22
申请人： DSM N.V. , DSM BIOTECH GMBH , JOHNS HOPKINS UNIVERSITY , TOWNSEND, Craig, A. , GUNSIOR, Michele , MÜLLER, Ulrike , ASSEMA VAN, Friso, Bernard, Jan , SONKE, Theodorus
发明人： TOWNSEND, Craig, A. , GUNSIOR, Michele , MÜLLER, Ulrike , ASSEMA VAN, Friso, Bernard, Jan , SONKE, Theodorus
IPC分类号： C12N15/53
摘要： A new fermentative process for the preparation of D- p -hydroxyphenylglycine (D-HPG) or D-henylglycine (D-PG) in enantiomerically pure form is disclosed. Precursors for the formation of D-HPG and D-PG are withdrawn form the common aromatic amino acid pathway, converted to p -hydroxyphenylglyoxylate or phenylglyoxylate, and are finally converted to D-HPG or D-PG by the action of a stero-inverting D-aminotransferase.

6. 发明申请

WO2004015094A1 PROCESS FOR THE PREPARATION OF L-3,4-DIHYDROXYPHENYLALANINE BY AEROBIC FERMENTATION OF A MICROORGANISM 审中-公开
标题翻译：通过微生物的微生物发酵制备L-3,4-二羟基苯甲酰胺的方法
公开(公告)号：WO2004015094A1
公开(公告)日：2004-02-19
申请号：PCT/EP2003/008507
申请日：2003-07-31
申请人： DSM IP ASSETS B.V. , DSM BIOTECH GMBH , KRÄMER, Marco , KREMER-MÜSCHEN, Susanne , WUBBOLTS, Marcel, Gerhardus
发明人： KRÄMER, Marco , KREMER-MÜSCHEN, Susanne , WUBBOLTS, Marcel, Gerhardus
IPC分类号： C12N9/02
CPC分类号： C12N9/0071 , C12P13/225
摘要： The invention relates to a process for the preparation of L-3,4-dihydroxyphenylalanine wherein L-3,4,-dihydroxyphenylalanine is produced in a fermentation medium by aerobic fermentation of a recombinant microorganism having L-tyrosine-3-hydroxy-mono-oxygenase activity and at least the metabolic pathways: glycolysis, pentose phosphate pathway, aromatic amino acid pathway, or derivative pathways thereof, which process comprises (i) a growth phase and a production phase, wherein L-3,4-dihydroxy-phenylalanine is produced in the fermentation medium, and 10 (ii) a downstream processing phase, and in which process L-3,4-dihydroxy phenylalanine is produced from a carbon source and the pH is in the range of from 1 to 7 during at least part of the production phase and/or downstream processing phase.
摘要翻译：本发明涉及一种制备L-3,4-二羟基苯丙氨酸的方法，其中在发酵培养基中通过需氧发酵具有L-酪氨酸-3-羟基 - 单 - 加氧酶活性和至少代谢途径：糖酵解，戊糖磷酸途径，芳香族氨基酸途径或其衍生途径，该方法包括（i）生长期和生产阶段，其中L-3,4-二羟基 - 苯丙氨酸是在发酵培养基中产生，并且10（ii）下游加工阶段，并且其中由碳源生产L-3,4-二羟基苯丙氨酸，并且在至少部分的pH范围为1至7的范围内的生产阶段和/或下游处理阶段。

7. 发明申请

WO2005118829A3 BIOSYNTHETIC PRODUCTION OF 4-AMINO-4-DEOXYCHORISMATE (ADC) AND [3R,4R]-4-AMINO-3-HYDROXYCYCLOHEXA-1,5-DIENE-1-CARBOXYLIC ACID (3,4-CHA) 审中-公开
标题翻译： 4-氨基-4-脱氧木糖（ADC）和[3R，4R] -4-氨基-3-羟基环己基-1,5-二苯基-1-羧酸（3,4-CHA）的生产生产
公开(公告)号：WO2005118829A3
公开(公告)日：2006-06-15
申请号：PCT/EP2005005988
申请日：2005-06-02
申请人： DSM IP ASSETS BV , DSM BIOTECH GMBH , FORSCHUNGSZENTRUM JUELICH GMBH , WUBBOLTS MARCEL GERHARDUS , BOVENBERG ROELOF ARY LANS , SPRENGER GEORG , BONGAERTS JOHANNES JOSEF , KOZAK STEFAN , MUELLER MICHAEL
发明人： WUBBOLTS MARCEL GERHARDUS , BOVENBERG ROELOF ARY LANS , SPRENGER GEORG , BONGAERTS JOHANNES JOSEF , KOZAK STEFAN , MUELLER MICHAEL
IPC分类号： C12P13/00 , C07C229/48 , C12N1/14 , C12N1/20 , C12P7/42 , C12P13/04
CPC分类号： C12P13/04
摘要： The invention relates to a process for the biosynthetic production of 4-amino-4-deoxychorismate (ADC) performed fermentativelyin vivo with a 4-amino-4-deoxychorismate synthase, preferably a PabAB bipartite protein (which may be a fusion protein), at an increased level of activity, thereby obtaining a broth comprising ADC and 4-amino-4-deoxyprephenate (ADP), that are recovered. The invention also relates to a further process of converting the ADP into p-aminophenylalanine. The invention, moreover relates to biosynthetic production of [3R,4R]-4-amino-3-hydroxycyclohexa-1,5-diene-1-carboxylic acid (3,4-CHA), by concerted action of such 4-amino-4-deoxychorismate synthase and of an enzyme capable of converting isochorismate into [5S,6S]-5,6 dihydroxycyclohexa-1,3-diene-1carboxylic acid (2,3-CHD), preferably a phenazine biosynthesis protein PhzD, including recovery of 3,4-CHA. The invention also relates to expression vectors and host cells for use in any of such processes. The invention further relates to the use of 3,4-CHA as a catalytically active product, in particular as a chiral catalyst. And the invention finally relates to synthesis of oseltamivir phosphate from 3,4-CHA.
摘要翻译：本发明涉及用4-氨基-4-脱氧激酶合酶，优选PabAB二分蛋白（可以是融合蛋白）发酵体内生物合成生产4-氨基-4-脱氧胸腺嘧啶（ADC）的方法，增加活性水平，从而获得包含ADC和4-氨基-4-脱氧降钙素（ADP）的肉汤，其被回收。本发明还涉及将ADP转化为对氨基苯丙氨酸的另一种方法。本发明还涉及通过这种4-氨基-3-羟基环己基-1,5-二烯-1-羧酸（3,4-CHA）的共同作用生产合成[3R，4R] -4-氨基-3-羟基环己-1,5-二烯-1-羧酸（3,4-CHA） 4-脱氧胸苷酸合酶和能够将异草酸转化为[5S，6S] -5,6二羟基环己-1,3-二烯-1-羧酸（2,3-CHD），优选吩嗪生物合成蛋白PhzD的酶的酶，包括回收的3,4-CHA。本发明还涉及用于任何这些方法的表达载体和宿主细胞。本发明还涉及3,4-CHA作为催化活性产物，特别是作为手性催化剂的用途。本发明最终涉及从3,4-CHA合成磷酸奥司他韦。

8. 发明申请

WO2003048374A1 PROCESS FOR THE PRODUCTION OF AN AROMATIC AMINO ACID METABOLITE OR DERIVATIVE THEREOF 审中-公开
标题翻译：制备芳族氨基酸代谢物或其衍生物的方法
公开(公告)号：WO2003048374A1
公开(公告)日：2003-06-12
申请号：PCT/NL2002/000796
申请日：2002-12-05
申请人： DSM N.V. , DSM BIOTECH GMBH , FORSCHUNGSZENTRUM JÜLICH GMBH , GERIGK, Marc, Robert , RAEVEN, Leon, Jean, Renier, Maria , SPRENGER, Georg , TAKORS, Ralf
发明人： GERIGK, Marc, Robert , RAEVEN, Leon, Jean, Renier, Maria , SPRENGER, Georg , TAKORS, Ralf
IPC分类号： C12P13/22
CPC分类号： C12P7/22 , C12P13/22
摘要： The invention relates to a process for the production of an aromatic amino acid metabolite or derivative thereof by aerobic fermentation of Escherichia coli , which fermentation comprises a growth and a production phase and in which fermentation glucose and L-tyrosine are controlled, wherein during at least part of the production phase, the glucose concentration in the fermentation medium is controlled within the range of 1-20 g/L and the L-tyrosine concentration in the fermentation medium is controlled below 36 mg/L.
摘要翻译：本发明涉及通过大肠杆菌的好氧发酵生产芳香族氨基酸代谢物或其衍生物的方法，该发酵包括生长和生产阶段，其中控制发酵葡萄糖和L-酪氨酸，其中至少在发酵培养基的一部分，发酵培养基中的葡萄糖浓度控制在1-20g / L的范围内，发酵培养基中的L-酪氨酸浓度控制在36mg / L以下。

9. 发明申请

WO0234921A2 FERMENTATIVE PRODUCTION OF D-p-HYDROXYPHENYLGLYCINE AND D-PHENYLGLYCINE 审中-公开
标题翻译： D-对羟基苯乙酸和D-苯乙醇的生产
公开(公告)号：WO0234921A2
公开(公告)日：2002-05-02
申请号：PCT/NL0100772
申请日：2001-10-22
申请人： DSM NV , DSM BIOTECH GMBH , UNIV JOHNS HOPKINS , TOWNSEND CRAIG A , GUNSIOR MICHELE , MUELLER ULRIKE , ASSEMA VAN FRISO BERNARD JAN , SONKE THEODORUS
发明人： TOWNSEND CRAIG A , GUNSIOR MICHELE , MUELLER ULRIKE , ASSEMA VAN FRISO BERNARD JAN , SONKE THEODORUS
IPC分类号： C12N1/21 , C12N9/10 , C12N15/52 , C12N15/53 , C12N15/54 , C12P21/02 , C12N9/02 , C12N9/04 , C12N15/63 , C12P13/04 , C12P41/00
CPC分类号： C12N15/52 , C12N9/1096 , C12P21/02
摘要： A new fermentative process for the preparation of D-p-hydroxyphenylglycine (D-HPG) or D-henylglycine (D-PG) in enantiomerically pure form is disclosed. Precursors for the formation of D-HPG and D-PG are withdrawn form the common aromatic amino acid pathway, converted to p-hydroxyphenylglyoxylate or phenylglyoxylate, and are finally converted to D-HPG or D-PG by the action of a stero-inverting D-aminotransferase.
摘要翻译：公开了以对映体纯的形式制备D-对羟基苯基甘氨酸（D-HPG）或D-丁炔基甘氨酸（D-PG）的新的发酵方法。用于形成D-HPG和D-PG的前体从共同的芳香族氨基酸途径中取出，转化为对羟基苯基乙醛酸酯或苯基乙醛酸酯，最后通过反相反应的作用转化为D-HPG或D-PG D-氨基转移酶。

10. 发明专利

DK1499411T3 未知
公开(公告)号：DK1499411T3
公开(公告)日：2007-10-22
申请号：DK03727226
申请日：2003-05-02
申请人： DSM BIOTECH GMBH
发明人： RUEFFER NICOLE , WANDREY CHRISTIAN , TAKORS RALF
IPC分类号： B01D11/04 , C12P13/04
摘要： The present invention relates to a process for integrated removal of one or more organic substances present in an aqueous bioprocess mixture, which comprise at least one positively charged and/or chargeable nitrogen-containing group, by means of reactive extraction in at least one step, in which process at least one liquid-liquid centrifuge is used for said reactive extraction and said organic substances are re-extracted from the extractant into an aqueous phase. More specifically, the bioprocess mixture directed to the liquid-liquid centrifuges is rendered cell-free and protein-free, before being directed into the first centrifuge. Furthermore, the invention relates to such a process in which an aqueous bioprocess mixture is continuously removed from a bioreactor, led, with an extractant, into a liquid-liquid centrifuge, extracted by means of said extractant, with an organic phase being obtained which contains the substance to be extracted from the fermentation mixture. Said substance may be re-extracted in a cycle via a second liquid-liquid centrifuge, resulting subsequently in a concentrated aqueous solution of the extracted substance, and the organic phase being recycled into the first liquid-liquid centrifuge.

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