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1. 发明申请

US20110045549A1 POLYNUCLEOTIDES ENCODING A FEEDBACK RESISTANT ASPARTOKINASE FROM CORYNEBACTERIUM 有权
标题翻译：从CORYNEBACTERIUM编码反馈抗生素的多核苷酸
公开(公告)号：US20110045549A1
公开(公告)日：2011-02-24
申请号：US12772538
申请日：2010-05-03
申请人： Paul D. Hanke , Lhing-Yew Li-D'Elia , Holly J. Walsh , Corey M. Crafton , P. John Rayapati
发明人： Paul D. Hanke , Lhing-Yew Li-D'Elia , Holly J. Walsh , Corey M. Crafton , P. John Rayapati
IPC分类号： C12P13/08 , C12N9/12 , C12N15/63 , C12N1/21 , C12N15/77 , C12N9/88 , C07K14/34 , C07H21/00
CPC分类号： C12N9/1217 , C07K14/34 , C07K14/345 , C12N9/88 , C12N15/52 , C12P13/04 , C12P13/08
摘要： The invention provides methods to increase the production of an amino acid from Corynebacterium species by way of the amplification of amino acid biosynthetic pathway genes in a host cell chromosome. The invention also provides novel processes for the production of an amino acid by way of the amplification of amino acid biosynthetic pathway genes in a host cell chromosome and/or by increasing promoter strength. In a preferred embodiment, the invention provides processes to increase the production of L-lysine in Corynebacterium glutamicum by way of the amplification of L-lysine biosynthetic pathway genes in a host cell chromosome. The invention also provides novel isolated nucleic acid molecules for L-lysine biosynthetic pathway genes of Corynebacterium glutamicum.
摘要翻译：本发明提供了通过在宿主细胞染色体中扩增氨基酸生物合成途径基因来增加棒状杆菌属物种的氨基酸生产的方法。本发明还提供了通过在宿主细胞染色体中扩增氨基酸生物合成途径基因和/或通过增加启动子强度来生产氨基酸的新方法。在优选的实施方案中，本发明提供了通过在宿主细胞染色体中扩增L-赖氨酸生物合成途径基因来增加谷氨酸棒杆菌中L-赖氨酸生产的方法。本发明还提供了用于谷氨酸棒杆菌的L-赖氨酸生物合成途径基因的新型分离的核酸分子。

2. 发明授权

US06927046B1 Increased lysine production by gene amplification using coryneform bacteria 有权
标题翻译：通过使用棒状细菌的基因扩增增加赖氨酸生产
公开(公告)号：US06927046B1
公开(公告)日：2005-08-09
申请号：US09722441
申请日：2000-11-28
申请人： Paul D. Hanke , Lhing-Yew Li-D'Elia , Holly J. Walsh , Corey M. Crafton , P. John Rayapati
发明人： Paul D. Hanke , Lhing-Yew Li-D'Elia , Holly J. Walsh , Corey M. Crafton , P. John Rayapati
IPC分类号： C12N15/09 , C07K14/34 , C07K14/345 , C12N1/21 , C12N9/12 , C12N9/88 , C12N15/52 , C12N15/54 , C12P13/04 , C12P13/08 , C12R1/13
CPC分类号： C12N9/1217 , C07K14/34 , C07K14/345 , C12N9/88 , C12N15/52 , C12P13/04 , C12P13/08
摘要： The invention provides methods to increase the production of an amino acid from Corynebacterium species by way of the amplification of amino acid biosynthetic pathway genes in a host cell chromosome. In a preferred embodiment, the invention provides methods to increase the production of L-lysine in Corynebacterium glutamicum by way of the amplification of L-lysine biosynthetic pathway genes in a host cell chromosome. The invention also provides novel processes for the production of an amino acid by way of the amplification of amino acid biosynthetic pathway genes in a host cell chromosome and/or by increasing promoter strength. In a preferred embodiment, the invention provides processes to increase the production of L-lysine in Corynebacterium glutamicum by way of the amplification of L-lysine biosynthetic pathway genes in a host cell chromosome. The invention also provides novel isolated nucleic acid molecules for L-lysine biosynthetic pathway genes of Corynebacterium glutamicum such as a naturally occurring, feedback-sensitive form of aspartokinase (ask) resulting from a threonine to isoleucine mutation at amino acid residue 380 in the ask gene of ATCC 21529, aspartate-semialdehyde dehydrogenase (asd), dihydrodipicolinate synthase (dapA), dihydrodipicolinate reductase (dapB), diaminopimelate dehydrogenase (ddh), and diaminopimelate decarboxylase (lysA).
摘要翻译：本发明提供了通过在宿主细胞染色体中扩增氨基酸生物合成途径基因来增加棒状杆菌属物种的氨基酸生产的方法。在优选的实施方案中，本发明提供了通过在宿主细胞染色体中扩增L-赖氨酸生物合成途径基因来增加谷氨酸棒杆菌中L-赖氨酸生产的方法。本发明还提供了通过在宿主细胞染色体中扩增氨基酸生物合成途径基因和/或通过增加启动子强度来生产氨基酸的新方法。在优选的实施方案中，本发明提供了通过在宿主细胞染色体中扩增L-赖氨酸生物合成途径基因来增加谷氨酸棒杆菌中L-赖氨酸生产的方法。本发明还提供了用于谷氨酸棒杆菌的L-赖氨酸生物合成途径基因的新型分离的核酸分子，例如在询问基因中的氨基酸残基380处的苏氨酸至异亮氨酸突变引起的天然存在的反馈敏感形式的天冬氨酸激酶（ask）（dapA），二氢吡啶二羧酸还原酶（dapB），二氨基庚二酸脱氢酶（ddh）和二氨基庚二酸脱羧酶（lysA）的天冬氨酸 - 半醛脱氢酶（asd），二氢吡啶二羧酸合酶（dapA）。

3. 发明授权

US06919190B2 Regulation of carbon assimilation 有权
标题翻译：碳同化调控
公开(公告)号：US06919190B2
公开(公告)日：2005-07-19
申请号：US09978698
申请日：2001-10-18
申请人： P. John Rayapati , Corey M. Crafton
发明人： P. John Rayapati , Corey M. Crafton
IPC分类号： C12N9/88 , C12P13/08 , C12P13/04 , C07H21/04 , C12N1/20
CPC分类号： C12N9/88 , C12P13/08
摘要： The present invention provides a method of increasing the productivity of a microorganism by improving the assimilation of carbon dioxide. Specifically, the invention provides a polypeptide having phosphoenolpyruvate carboxylase activity which does not require acetyl coenzyme A for activation and is desensitized to feedback inhibition by aspartic acid, and to genes coding for this polypeptide. A gene encoding a PEP carboxylase that is not regulated by acetyl-CoA or aspartic acid can improve carbon flow from the three carbon intermediate PEP to the four carbon intermediate OAA, contribute to compounds derived from OAA, and increase amino acid biosynthesis. The invention further provides recombinant DNA molecules containing these genes, bacteria transformed with these genes, and a method of producing amino acids using the transformed bacteria.
摘要翻译：本发明提供了通过改善二氧化碳的同化来提高微生物的生产率的方法。具体地，本发明提供了具有磷酸烯醇丙酮酸羧化酶活性的多肽，其不需要乙酰辅酶A用于激活，并且对天冬氨酸的反馈抑制脱敏，以及编码该多肽的基因。编码不受乙酰辅酶A或天冬氨酸调节的PEP羧化酶的基因可以改善从三碳中间体PEP到四碳中间体OAA的碳流动，有助于衍生自OAA的化合物，并增加氨基酸生物合成。本发明还提供含有这些基因的重组DNA分子，用这些基因转化的细菌，以及使用转化细菌产生氨基酸的方法。

4. 发明授权

US07141388B2 Nucleotide sequences for transcriptional regulation in corynebacterium glutamicum 有权
公开(公告)号：US07141388B2
公开(公告)日：2006-11-28
申请号：US09987763
申请日：2001-11-15
申请人： Corey M. Crafton , P. John Rayapati
发明人： Corey M. Crafton , P. John Rayapati
IPC分类号： C12P21/06 , C12N15/00 , C12N1/20 , C12N15/74 , C07H21/04
CPC分类号： C12N15/77 , C07K14/34 , C07K2319/00
摘要： The invention relates to isolated polynucleotides from Corynebacterium glutamicum which are useful in the regulation of gene expression. In particular, the invention relates to isolated polynucleotides comprising C.glutamicum promoters which may be used to regulate, i.e., either increase or decrease, gene expression. In certain embodiments, isolated promoter sequences of the present invention regulate gene expression through the use of exogenous or endogenous induction. The invention further provides recombinant vectors and recombinant cells comprising isolated polynucleotides of the present invention, preferably in operable association with heterologous genes. Also provided are methods of regulating bacterial gene expression comprising growth of a recombinant cell of the present invention. In particular, the present invention provides methods to regulate genes involved in amino acid production comprising growth of a recombinant cell of the present invention. In certain embodiments, the present invention provides methods of regulating gene expression in bacteria, particularly Corynebacterium species, especially of the genus Corynebacterium, comprising fermentation growth of a recombinant cell of the present invention, where metabolite concentrations, temperature, or oxygen levels are manipulated to regulate gene expression.

5. 发明授权

US07635579B2 Metabolic engineering of amino acid production 有权
标题翻译：氨基酸生产代谢工程
公开(公告)号：US07635579B2
公开(公告)日：2009-12-22
申请号：US11978908
申请日：2007-10-30
申请人： P. John Rayapati , Corey M. Crafton
发明人： P. John Rayapati , Corey M. Crafton
IPC分类号： C12P13/04 , C12P13/08 , C12N15/74 , C12N1/20
CPC分类号： C12N15/01 , C12N9/88 , C12P13/04 , C12P13/08
摘要： The present invention is directed towards the fermentative production of amino acids, providing microorganisms, methods and processes useful therefor. Microorganisms of the invention are capable of converting glucose to amino acids other than L-isoleucine, L-leucine and L-valine with greater efficiency than the parent strain. The efficiency of conversion may be quantified by the formula: [(g amino acid produced/g dextrose consumed)*100]=% Yield and expressed as yield from dextrose. The invention provides microorganisms that are made auxotrophic or bradytrophic for the synthesis of one or more branched chain amino acids by mutagenesis and selected for their ability to produce higher percent yields of the desired amino acid than the parental strain. Preferred microorganisms are Corynebacterium, Brevibacterium or Escherichia coli producing L-lysine. Mutagenesis is performed by classical techniques or through rDNA methodology. Methods of the invention are designed to increase the production of an amino acid by mutagenizing a parental strain, selecting cells auxotrophic or bradytrophic for the synthesis of one or more branched chain amino acids and selecting branched chain amino acid auxotrophs or bradytrophs that produce a higher percent yield from dextrose of the desired amino acid than the parental strain. Processes of the invention are designed for the production an amino acid comprising culturing in a medium a microorganism obtained by mutagenizing a parent strain to be auxotrophic or bradytrophic for branched chain amino acid synthesis and selecting variants that are capable of converting glucose to amino acids other than L-isoleucine, L-leucine and L-valine with greater efficiency than the parent strain.
摘要翻译：本发明涉及氨基酸的发酵生产，提供微生物，对其有用的方法和方法。本发明的微生物能够以比亲本菌株更高的效率将葡萄糖转化成L-异亮氨酸，L-亮氨酸和L-缬氨酸以外的氨基酸。转化效率可以通过以下公式进行定量：[（g氨基酸产生/ g葡萄糖消耗）* 100] =％产率并以葡萄糖的收率表示。本发明提供了通过诱变合成一种或多种支链氨基酸产生营养缺陷型或营养缺陷型微生物，并且选择其产生比亲本菌株更高产率的所需氨基酸的能力。优选的微生物是产生L-赖氨酸的棒状杆菌，短杆菌或大肠杆菌。诱变通过经典技术或通过rDNA方法进行。本发明的方法被设计成通过诱变亲本菌株来增加氨基酸的产生，选择营养缺陷型或营养型细胞用于合成一种或多种支链氨基酸并选择产生更高百分比的支链氨基酸营养缺陷型或营养缺陷型来自所需氨基酸的葡萄糖的产量高于亲本菌株。本发明的方法被设计用于生产氨基酸，其包括在培养基中培养通过将亲本菌株诱变为营养缺陷型或营养型的支链氨基酸合成获得的微生物，并选择能够将葡萄糖转化成除了氨基酸以外的氨基酸的变体 L-异亮氨酸，L-亮氨酸和L-缬氨酸，其效力高于亲本菌株。

6. 发明授权

US07323321B2 Metabolic engineering of amino acid production 有权
标题翻译：氨基酸生产代谢工程
公开(公告)号：US07323321B2
公开(公告)日：2008-01-29
申请号：US11320139
申请日：2005-12-28
申请人： P. John Rayapati , Corey M. Crafton
发明人： P. John Rayapati , Corey M. Crafton
IPC分类号： C12P13/04 , C12P13/08 , C12N15/74 , C12N1/20
CPC分类号： C12N15/01 , C12N9/88 , C12P13/04 , C12P13/08
摘要： The present invention is directed towards the fermentative production of amino acids, providing microorganisms, methods and processes useful therefor. Microorganisms of the invention are capable of converting glucose to amino acids other than L-isoleucine, L-leucine and L-valine with greater efficiency than the parent strain. The efficiency of conversion may be quantified by the formula: [(g amino acid produced/g dextrose consumed)*100]=% Yield and expressed as yield from dextrose. The invention provides microorganisms that are made bradytrophic for the synthesis of valine by mutagenesis.
摘要翻译：本发明涉及氨基酸的发酵生产，提供微生物，对其有用的方法和方法。本发明的微生物能够以比亲本菌株更高的效率将葡萄糖转化成L-异亮氨酸，L-亮氨酸和L-缬氨酸以外的氨基酸。转化效率可以通过以下公式进行定量：[（g氨基酸产生/ g葡萄糖消耗）* 100] =％产率并以葡萄糖的收率表示。本发明提供通过诱变制备缬氨酸营养不良的微生物。

7. 发明授权

US06599732B1 Regulation of carbon assimilation 有权
公开(公告)号：US06599732B1
公开(公告)日：2003-07-29
申请号：US09606312
申请日：2000-06-29
申请人： P. John Rayapati , Corey M. Crafton
发明人： P. John Rayapati , Corey M. Crafton
IPC分类号： C12N988
CPC分类号： C12N9/88 , C12P13/08
摘要： The present invention provides a method of increasing the productivity of a microorganism by improving the assimilation of carbon dioxide. Specifically, the invention provides a polypeptide having phosphoenolpyruvate carboxylase activity which does not require acetyl coenzyme A for activation and is desensitized to feedback inhibition by aspartic acid, and to genes coding for this polypeptide. A gene encoding a PEP carboxylase that is not regulated by acetyl-CoA or aspartic acid can improve carbon flow from the three carbon intermediate PEP to the four carbon intermediate OAA, contribute to compounds derived from OAA, and increase amino acid biosynthesis. The invention further provides recombinant DNA molecules containing these genes, bacteria transformed with these genes, and a method of producing amino acids using the transformed bacteria.

8. 发明申请

US20090042264A1 COMPOSITIONS AND METHODS FOR MANIPULATING CARBON FLUX IN CELLS 有权
标题翻译：用于在细胞中操作碳通量的组合物和方法
公开(公告)号：US20090042264A1
公开(公告)日：2009-02-12
申请号：US12195869
申请日：2008-08-21
申请人： Beth Fatland-Bloom , Gyan Rai , P. John Rayapati , Nyerhovwo John Tonukari
发明人： Beth Fatland-Bloom , Gyan Rai , P. John Rayapati , Nyerhovwo John Tonukari
IPC分类号： C12P7/56 , C12N15/11 , C12N15/00 , C12P7/20 , C12P7/46 , C12N1/20 , C12N1/19
CPC分类号： C12P7/56 , C12N9/93 , C12P7/20 , C12P7/46
摘要： Nucleotide sequences and genetic constructs that can be used to regulate genes encoding enzymes that change carbon flux through metabolic pathways that lead to lactic acid or fumarate production in a host cell, such as a R. oryzae cell, are provided. Methods of manipulating carbon flux in a cell also are provided.
摘要翻译：提供了可用于调节编码通过代谢途径转化碳通量的基因的基因组序列和遗传构建体，所述代谢途径导致宿主细胞例如米曲霉细胞中的乳酸或富马酸盐的产生。还提供了操作细胞中碳通量的方法。

9. 发明申请

US20100112651A1 COMPOSITIONS AND METHODS FOR MANIPULATING CARBON FLUX IN CELLS 有权
标题翻译：用于在细胞中操作碳通量的组合物和方法
公开(公告)号：US20100112651A1
公开(公告)日：2010-05-06
申请号：US12409072
申请日：2009-03-23
申请人： Beth Fatland-Bloom , P. John Rayapati , Nyerhovwo John Tonukari
发明人： Beth Fatland-Bloom , P. John Rayapati , Nyerhovwo John Tonukari
IPC分类号： C12P7/56 , C07H21/00 , C07H21/02 , C12N15/74 , C12N1/21 , C12N1/19 , C12N1/15 , C12P7/46 , C12P7/20
CPC分类号： C12P7/46 , C12N9/93 , C12P7/20 , C12P7/56 , C12Y604/01001
摘要： Nucleotide and protein sequences that encode enzymes that change carbon flux through metabolic pathways that lead to lactic acid or fumarate production in a host cell, such as a R. oryzae cell, are provided. Methods of manipulating carbon flux in a cell also are provided.
摘要翻译：提供编码通过代谢途径改变碳通量的酶的核苷酸和蛋白质序列，其导致宿主细胞如米曲霉细胞中的乳酸或富马酸盐的产生。还提供了操作细胞中碳通量的方法。

10. 发明授权

US07435168B2 Compositions and methods for manipulating carbon flux in cells 有权
标题翻译：用于操纵细胞中碳通量的组合物和方法
公开(公告)号：US07435168B2
公开(公告)日：2008-10-14
申请号：US11334713
申请日：2006-01-17
申请人： Beth Fatland-Bloom , Gyan Rai , P. John Rayapati , Nyerhovwo John Tonukari
发明人： Beth Fatland-Bloom , Gyan Rai , P. John Rayapati , Nyerhovwo John Tonukari
IPC分类号： C12P21/06 , C12P19/34 , C12P7/56 , C12P7/46 , C12N1/20 , C12N1/00 , C12N15/74
CPC分类号： C12P7/56 , C12N9/93 , C12P7/20 , C12P7/46
摘要： Nucleotide sequences and genetic constructs that can be used to regulate genes encoding enzymes that change carbon flux through metabolic pathways that lead to lactic acid or fumarate production in a host cell, such as a R. oryzae cell, are provided. Methods of manipulating carbon flux in a cell also are provided.
摘要翻译：提供了可用于调节编码通过代谢途径转化碳通量的基因的基因组序列和遗传构建体，所述代谢途径导致宿主细胞例如米曲霉细胞中的乳酸或富马酸盐的产生。还提供了操作细胞中碳通量的方法。

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