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1. 发明申请

US20090246835A1 L-AMINO ACID-PRODUCING BACTERIUM AND A METHOD FOR PRODUCING AN L-AMINO ACID 有权
标题翻译： L-氨基酸生产细菌和一种生产L-氨基酸的方法
公开(公告)号：US20090246835A1
公开(公告)日：2009-10-01
申请号：US12055438
申请日：2008-03-26
申请人： Shintaro Iwatani , Akira Imaizumi , Yoshihiro Usuda , Kazuhiko Matsui
发明人： Shintaro Iwatani , Akira Imaizumi , Yoshihiro Usuda , Kazuhiko Matsui
IPC分类号： C12P13/22 , C12P13/04 , C12P13/08
CPC分类号： C07K14/245 , C12P13/04 , C12P13/08 , C12P13/22
摘要： An L-amino acid is produced by culturing a microorganism of the family Enterobacteriaceae which has the ability to produce an L-amino acid and which has been modified so as to increase the expression of the evgA gene, the gadE gene, and/or the ydeO gene. These genes encode a transcription factor involved in the EvgAS two-component system regulon. The culture takes place in a medium, and the L-amino acid is collected from the medium or cells.
摘要翻译： L-氨基酸是通过培养具有产生L-氨基酸的能力的肠杆菌科的微生物而产生的，该微生物被修饰以增加evgA基因，gadE基因和/或 ydeO基因。这些基因编码参与EvgAS双组分系统调节子的转录因子。培养物在培养基中进行，从培养基或细胞收集L-氨基酸。

2. 发明授权

US09644009B2 L-amino acid-producing bacterium and a method for producing an L-amino acid 有权
公开(公告)号：US09644009B2
公开(公告)日：2017-05-09
申请号：US12055438
申请日：2008-03-26
申请人： Shintaro Iwatani , Akira Imaizumi , Yoshihiro Usuda , Kazuhiko Matsui
发明人： Shintaro Iwatani , Akira Imaizumi , Yoshihiro Usuda , Kazuhiko Matsui
IPC分类号： C12P13/04 , C07K14/245 , C12P13/08 , C12P13/22
CPC分类号： C07K14/245 , C12P13/04 , C12P13/08 , C12P13/22
摘要： An L-amino acid is produced by culturing a microorganism of the family Enterobacteriaceae which has the ability to produce an L-amino acid and which has been modified so as to increase the expression of the evgA gene, the gadE gene, and/or the ydeO gene. These genes encode a transcription factor involved in the EvgAS two-component system regulon. The culture takes place in a medium, and the L-amino acid is collected from the medium or cells.

3. 发明授权

US08510054B2 Intracellular metabolic flux analysis method using substrate labeled with isotope 失效
标题翻译：使用同位素标记的底物的细胞内代谢通量分析方法
公开(公告)号：US08510054B2
公开(公告)日：2013-08-13
申请号：US11048923
申请日：2005-02-03
申请人： Shintaro Iwatani , Yoshihiro Usuda , Kazuhiko Matsui
发明人： Shintaro Iwatani , Yoshihiro Usuda , Kazuhiko Matsui
IPC分类号： G01N33/48 , C12Q1/00
CPC分类号： G01N33/5091 , C12Q1/00 , G06F19/12
摘要： A method for intracellular metabolic flux analysis comprising (a) culturing cells in a medium not containing any isotope-labeled substrate to a target phase of the metabolic flux analysis, (b) adding an isotope-labeled substrate to the medium, and continuing culture and collecting samples from the medium in time course, (c) measuring isotope distribution in an intracellular metabolite contained in the samples collected in time course, (d) performing a regression analysis for measured data and calculating an isotope distribution ratio in a steady state, and (e) analyzing a metabolic flux of the cultured cells by using the calculated isotope distribution ratio.
摘要翻译：一种用于细胞内代谢通量分析的方法，包括（a）在不含任何同位素标记的底物的培养基中将细胞培养至代谢通量分析的目标阶段，（b）将同位素标记的底物加入到培养基中，在时间过程中从培养基中收集样品，（c）测量在时间过程中收集的样品中所含的细胞内代谢物中的同位素分布，（d）对稳定状态进行测量数据和计算同位素分布比的回归分析，以及（e）使用计算出的同位素分布比分析培养细胞的代谢通量。

4. 发明授权

US08030036B2 Method for producing L-lysine or L-threonine 有权
标题翻译：生产L-赖氨酸或L-苏氨酸的方法
公开(公告)号：US08030036B2
公开(公告)日：2011-10-04
申请号：US11877726
申请日：2007-10-24
申请人： Stephen Van Dien , Shintaro Iwatani , Yoshihiro Usuda , Kazuhiko Matsui , Yuta Nakai , Tomoko Suzuki , Mika Moriya , Yuichiro Tsuji , Takuji Ueda
发明人： Stephen Van Dien , Shintaro Iwatani , Yoshihiro Usuda , Kazuhiko Matsui , Yuta Nakai , Tomoko Suzuki , Mika Moriya , Yuichiro Tsuji , Takuji Ueda
IPC分类号： C12P13/08 , C12N9/04 , C12N1/21 , C07H21/04
CPC分类号： C12N9/0006 , C12P13/08
摘要： A bacterium belonging to the genus Escherichia which has an ability to produce L-lysine or L-threonine and which is modified so that a malic enzyme does not function normally in a cell, and a method for producing L-lysine or L-threonine, comprising culturing the bacterium in a medium to produce and cause accumulation of L-lysine or L-threonine, and collecting the L-lysine or L-threonine from the medium.
摘要翻译：属于埃希氏菌属的细菌，其具有产生L-赖氨酸或L-苏氨酸的能力，并且其被修饰以使得苹果酸在细胞中不起作用;以及L-赖氨酸或L-苏氨酸的制备方法，包括在培养基中培养细菌以产生并引起L-赖氨酸或L-苏氨酸的积累，并从培养基中收集L-赖氨酸或L-苏氨酸。

5. 发明申请

US20090239269A1 METHOD FOR PRODUCTION OF L-AMINO ACID 有权
标题翻译： L-氨基酸生产方法
公开(公告)号：US20090239269A1
公开(公告)日：2009-09-24
申请号：US12420934
申请日：2009-04-09
申请人： Yoshinori Tajima , Shintaro Iwatani , Yoshihiro Usuda , Kazuhiko Matsui
发明人： Yoshinori Tajima , Shintaro Iwatani , Yoshihiro Usuda , Kazuhiko Matsui
IPC分类号： C12P13/20 , C12P13/08
CPC分类号： C12N9/88 , C12P13/08 , C12P13/227
摘要： A bacterium which belongs to the Enterobacteriaceae family and has an ability to produce an L-amino acid such as L-lysine, L-threonine and L-tryptophan and is modified to enhance glutamic acid decarboxylase activity is cultured in a medium to produce and accumulate the L-amino acid in the medium or cells of the bacterium. Then, the L-amino acid is collected from the medium or the cells.
摘要翻译：属于肠杆菌科的具有生产L-赖氨酸，L-苏氨酸和L-色氨酸的L-氨基酸的能力并经过修饰以增强谷氨酸脱羧酶活性的细菌在培养基中培养以产生和积累细菌的培养基或细胞中的L-氨基酸。然后，从培养基或细胞收集L-氨基酸。

6. 发明授权

US07306933B2 Method for producing L-lysine or L-threonine 有权
标题翻译：生产L-赖氨酸或L-苏氨酸的方法
公开(公告)号：US07306933B2
公开(公告)日：2007-12-11
申请号：US11275437
申请日：2006-01-03
申请人： Stephen Van Dien , Shintaro Iwatani , Yoshihiro Usuda , Kazuhiko Matsui , Yuta Nakai , Tomoko Suzuki , Mika Moriya , Yuichiro Tsuji , Takuji Ueda
发明人： Stephen Van Dien , Shintaro Iwatani , Yoshihiro Usuda , Kazuhiko Matsui , Yuta Nakai , Tomoko Suzuki , Mika Moriya , Yuichiro Tsuji , Takuji Ueda
IPC分类号： C12P13/08 , C12N9/04 , C12N1/21 , C07H21/04
CPC分类号： C12N9/0006 , C12P13/08
摘要： A bacterium belonging to the genus Escherichia which has an ability to produce L-lysine or L-threonine and which is modified so that a malic enzyme does not function normally in a cell, and a method for producing L-lysine or L-threonine, comprising culturing the bacterium in a medium to produce and cause accumulation of L-lysine or L-threonine, and collecting the L-lysine or L-threonine from the medium.
摘要翻译：属于埃希氏菌属的细菌，其具有产生L-赖氨酸或L-苏氨酸的能力，并且其被修饰以使得苹果酸在细胞中不起作用;以及L-赖氨酸或L-苏氨酸的制备方法，包括在培养基中培养细菌以产生并引起L-赖氨酸或L-苏氨酸的积累，并从培养基中收集L-赖氨酸或L-苏氨酸。

7. 发明申请

US20070038419A1 Method for simulating a production process of a substance 审中-公开
标题翻译：模拟物质生产过程的方法
公开(公告)号：US20070038419A1
公开(公告)日：2007-02-15
申请号：US11501232
申请日：2006-08-09
申请人： Yoshihiro Usuda , Shintaro Iwatani , Kazuhiko Matsui
发明人： Yoshihiro Usuda , Shintaro Iwatani , Kazuhiko Matsui
IPC分类号： G06F17/10
CPC分类号： G16B5/00
摘要： A method for simulating a substance production process using cells based on a set of differential equations that represent intracellular metabolites and gene expression comprising including a specific growth rate of cells, expressed as a differential equation, in the set of differential equations; assigning values for parameters in the set of differential equations, wherein at least one of said parameters is represented as a growth rate factor; incorporating in the set of differential equations a formation rate for formation of a cell component, wherein said formation rate is represented as a growth rate factor; incorporating in the set of differential equations an inflow rate of a metabolite taken from outside of the cells and/or an outflow rate of a metabolite excreted out of the cells, wherein said inflow/outflow rates are represented as a growth rate factor; solving the set of differential equations; and generating data representative of the substance-production process.
摘要翻译：一种基于代表细胞内代谢物和基因表达的微分方程组，使用细胞模拟物质生产过程的方法，其包括在微分方程组中包含表示为微分方程的细胞的比生长速率; 为所述微分方程组中的参数分配值，其中所述参数中的至少一个被表示为增长率因子; 在微分方程组中并入形成细胞成分的形成速率，其中所述形成速率表示为生长速率因子; 在微分方程组中引入从细胞外部获取的代谢物的流入速率和/或从细胞排出的代谢物的流出速率，其中所述流入/流出速率表示为生长速率因子; 求解微分方程组; 并产生代表物质生产过程的数据。

8. 发明授权

US07555392B2 Method for determining metabolic flux 失效
公开(公告)号：US07555392B2
公开(公告)日：2009-06-30
申请号：US11095521
申请日：2005-04-01
申请人： Shintaro Iwatani , Stephen Van Dien , Yoshihiro Usuda , Kazuhiko Matsui
发明人： Shintaro Iwatani , Stephen Van Dien , Yoshihiro Usuda , Kazuhiko Matsui
IPC分类号： G06F7/00
CPC分类号： G06F19/12 , Y10S707/99943
摘要： A method for determining metabolic flux distribution of a cell, which comprises the steps of: 1) creating a stoichiometric matrix based on formulas of biochemical reactions from a substrate to a product, 2) computing a set of vectors of solutions existing in a solution space which the stoichiometric matrix may have, 3) selecting, from the computed set of vectors of solutions, maximum vectors whose vector elements corresponding to respective substances for which input values can be obtained are maximum, and 4) performing linear combination for the selected vectors in accordance with the following equations to obtain a vector representing metabolic flux distribution: S flux = ∑ i = 1 n ⁢ a i · p i · S ⁡ ( i ) + ∑ i = 1 n ⁢ b i · q i · A ⁡ ( i ) ( I ) ∑ i = 1 n ⁢ a i + ∑ i = 1 n ⁢ b i = 1 ( II ) Sflux: Vector representing metabolic flux distribution to be determined, S(i): Vector selected for substance for which inputted value can be obtained, A(i): Adjustment vector, a, b: Coefficients for linear combination (b may be 0), p: Coefficient for obtaining consistency with input value as numerical value, q: Coefficient of adjustment vector.

9. 发明申请

US20060154289A1 Method for determining metabolic flux affecting substance production 有权
标题翻译：测定代谢通量影响物质生产的方法
公开(公告)号：US20060154289A1
公开(公告)日：2006-07-13
申请号：US11337580
申请日：2006-01-24
申请人： Stephen Dien , Shintaro Iwatani , Yoshihiro Usuda , Kazuhiko Matsui , Takuji Ueda , Yuichiro Tsuji
发明人： Stephen Dien , Shintaro Iwatani , Yoshihiro Usuda , Kazuhiko Matsui , Takuji Ueda , Yuichiro Tsuji
IPC分类号： C12Q1/68 , C12Q1/18 , C12N1/21
CPC分类号： C12N9/0006 , C12P13/08
摘要： A metabolic flux affecting substance production using cells is determined by 1) creating a stoichiometric matrix based on formulas of biochemical reactions from a substrate through a desired produced substance, 2) selecting the same number of independent metabolic fluxes from all metabolic fluxes as the degree of freedom of the stoichiometric matrix as free fluxes, 3) creating a sufficient number of random combinations of the free fluxes for a statistical analysis and calculating a metabolic flux distribution from each created combination based on the stoichiometric matrix, 4) obtaining a regression equation including a minimum number of free fluxes that shows a correlation with substance production from the calculated metabolic flux distributions by a multivariate statistical analysis, and 5) determining at least one metabolic flux affecting substance production based on a coefficient in the obtained regression equation.
摘要翻译：影响使用细胞的物质生产的代谢通量是通过以下方法确定的：1）根据基质通过所需生产物质的生物化学反应的公式产生化学计量矩阵，2）从所有代谢通量中选择相同数量的独立代谢通量，作为自由通量的化学计量矩阵的自由度，3）产生足够数量的自由通量的随机组合用于统计分析，并且基于化学计量矩阵计算每个创建的组合的代谢通量分布，4）获得包括通过多变量统计分析从计算的代谢通量分布中显示与物质生产相关的最小数量的自由通量，以及5）基于所获得的回归方程中的系数确定影响物质生产的至少一种代谢通量。

10. 发明申请

US20060105322A1 Intracellular metabolic flux analysis method using substrate labeled with isotope 审中-公开
标题翻译：使用同位素标记的底物的细胞内代谢通量分析方法
公开(公告)号：US20060105322A1
公开(公告)日：2006-05-18
申请号：US11319104
申请日：2005-12-28
申请人： Shintaro Iwatani , Stephen Van Dien , Yoshihiro Usuda , Kazuhiko Matsui
发明人： Shintaro Iwatani , Stephen Van Dien , Yoshihiro Usuda , Kazuhiko Matsui
IPC分类号： C12Q1/00 , G06F19/00
CPC分类号： G16B5/00
摘要： A method for analyzing an intracellular metabolic flux comprising determining the intracellular metabolic flux from analytical values of cells cultured in a medium containing an isotope-labeled substrate as a carbon source on the basis of an intracellular metabolic flux model constructed for the intracellular metabolic flux to be analyzed, wherein (a) influence of an exchange reaction between an intracellular metabolite and a cell component produced by integration of the intracellular metabolite is considered, (b) uptake of a compound in a medium into cells, which compound is identical to an intracellular metabolite and unlabeled with an isotope, is considered, or (c) carbon dioxide used in a fixation reaction is assumed as carbon dioxide produced in a production reaction.
摘要翻译：一种用于分析细胞内代谢通量的方法，其包括基于在构建用于细胞内代谢通量的细胞内代谢通量模型的基础上，在含有同位素标记的底物作为碳源的培养基中培养的细胞的分析值来测定细胞内代谢通量分析，其中（a）细胞内代谢物与通过细胞内代谢物的整合产生的细胞成分之间的交换反应的影响被考虑，（b）将培养基中的化合物摄取到细胞中，该化合物与细胞内代谢物相同被认为是未标记的同位素，或（c）在定影反应中使用的二氧化碳被认为是在生产反应中产生的二氧化碳。

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