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公开(公告)号：US07132257B2
公开(公告)日：2006-11-07
申请号：US10886906
申请日：2004-07-08
申请人： Qiong Cheng , Pierre E. Rouviere , Wonchul Suh , Luan Tao
发明人： Qiong Cheng , Pierre E. Rouviere , Wonchul Suh , Luan Tao
CPC分类号： C12P23/00
摘要： A method for the in vivo bioconversion of cyclic carotenes having a β-ionone ring to the corresponding aryl carotene is provided. Gram negative host cells expressing a heterologous, codon-optimized gene encoding a carotene desaturase are grown in the presence of a suitable cyclic carotene substrate to effect the production of aromatic carotenoids.
摘要翻译： 提供了具有β-紫罗兰酮环到相应的芳基胡萝卜素的体内生物转化环状胡萝卜素的方法。 表达编码胡萝卜素去饱和酶的异源，密码子优化的基因的革兰氏阴性宿主细胞在合适的环状胡萝卜素底物的存在下生长，以产生芳香类胡萝卜素。
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公开(公告)号：US20120214202A1
公开(公告)日：2012-08-23
申请号：US13164990
申请日：2011-06-21
申请人： Ritu Bhalla , Qi Chen , Qiong Cheng , Neeraj Pandey , Pierre E. Rouviere , Kristin Ruebling-Jass , Annapurna Sachan
发明人： Ritu Bhalla , Qi Chen , Qiong Cheng , Neeraj Pandey , Pierre E. Rouviere , Kristin Ruebling-Jass , Annapurna Sachan
CPC分类号： C12P21/02 , C12N1/02 , C12N9/1025 , C12N15/70
摘要： Methods are provided to obtain recombinant microbial cells having at least one genetic modification that increase the buoyant density of a recombinant microbial cell or the buoyant density of inclusion bodies produced within a recombinant microbial cell. Exemplified are genetic modifications that increase the buoyant density of a recombinant microbial cell expressing heterologous peptides and polypeptides. Increasing expression of the genes ysaB, glyQ, glyS or a combination thereof within the recombinant microbial cell produces cells or inclusion bodies having higher buoyant density. A similar effect was achieved by decreasing or disrupting expression of the endogenous gltA gene. Increases in buoyant density render peptide production more efficient with respect to time and costs.
摘要翻译： 提供了获得具有增加重组微生物细胞的浮力密度或在重组微生物细胞内产生的包涵体的浮力密度的至少一种遗传修饰的重组微生物细胞的方法。 示例是增加表达异源肽和多肽的重组微生物细胞的浮力密度的遗传修饰。 在重组微生物细胞内增加基因ysaB，glyQ，glyS或其组合的表达产生具有较高浮力密度的细胞或包涵体。 通过减少或破坏内源性gltA基因的表达也可以获得类似的效果。 浮力密度的增加使肽生产在时间和成本方面更有效率。
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公开(公告)号：US07794979B2
公开(公告)日：2010-09-14
申请号：US12692665
申请日：2010-01-25
申请人： Qiong Cheng , Linda Jane Decarolis , Stephen R. Fahnestock , Tanja Maria Gruber , Lisa Diane Reiss , Pierre E. Rouviere
发明人： Qiong Cheng , Linda Jane Decarolis , Stephen R. Fahnestock , Tanja Maria Gruber , Lisa Diane Reiss , Pierre E. Rouviere
CPC分类号： C12P21/02 , C07K2319/50 , C12N9/90
摘要： Peptide tags, referred to here as inclusion body tags, are disclosed useful for the generation of insoluble fusion peptides. The fusion peptides comprise at least one inclusion body tag operably linked to a peptide of interest. Expression of the fusion peptide in a host cell results in a product that is insoluble and contained within inclusion bodies in the cell and/or cell lysate. The inclusion bodies may then be purified and the protein of interest may be isolated after cleavage from the inclusion body tag.
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14.发明申请公开(公告)号：US20090043075A1
公开(公告)日：2009-02-12
申请号：US12172395
申请日：2008-07-14
申请人： Albert W. Alsop , Qiong Cheng , Linda Jane Decarolis , Stephen R. Fahnestock , Tanja Maria Gruber , Pierre E. Rouviere
发明人： Albert W. Alsop , Qiong Cheng , Linda Jane Decarolis , Stephen R. Fahnestock , Tanja Maria Gruber , Pierre E. Rouviere
CPC分类号： C12P21/02 , C07K2319/00 , C12P21/06
摘要： The invention relates to the recombinant expression of a peptide of interest in the form of a fusion protein comprising a solubility tag. The fusion protein comprises at least two portions separated by a cleavable peptide sequence wherein one portion is devoid of cysteine residues and the second portion comprises an effective number of cross-linkable cysteine residues. After cell lysis and isolation of the fusion protein, the fusion protein is subsequently cleaved into a mixture of first and second portions. Oxidative cross-linking is used to selectively precipitate one of the two portions to facilitate simple and effective separation of the peptide of interest.
摘要翻译： 本发明涉及以包含溶解性标签的融合蛋白形式的目标肽的重组表达。 融合蛋白包含由可切割肽序列分开的至少两个部分，其中一部分缺少半胱氨酸残基，第二部分包含有效数目的可交联半胱氨酸残基。 细胞裂解和分离融合蛋白后，融合蛋白随后被切割成第一和第二部分的混合物。 氧化交联用于选择性沉淀两部分之一，以促进目的肽的简单和有效分离。
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公开(公告)号：US07232665B2
公开(公告)日：2007-06-19
申请号：US10735008
申请日：2003-12-12
申请人： Qiong Cheng , Pierre E. Rouviere , Luan Tao
发明人： Qiong Cheng , Pierre E. Rouviere , Luan Tao
摘要： Mutations in genes having no direct relationship to the carotenoid biosynthetic pathway have been found to increase carbon flux through that pathway. Complete disruption in the deaD, mreC, and yfhE genes were effective. Additionally where genes of the lower carotenoid pathway reside on a plasmid having either a p15A or pMB1 replicon, mutations in the thrS, rspA, rpoC, yjeR, and rhoL were found effective.
摘要翻译： 已经发现与类胡萝卜素生物合成途径没有直接关系的基因中的突变增加通过该途径的碳通量。 deaD，mreC和yfhE基因的完全破坏是有效的。 此外，较低类胡萝卜素途径的基因存在于具有p15A或pMB1复制子的质粒上，发现thrS，rspA，rpoC，yjeR和rhoL中的突变是有效的。
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16.发明申请公开(公告)号：US20100227361A1
公开(公告)日：2010-09-09
申请号：US12575550
申请日：2009-10-08
CPC分类号： C07K14/245 , C07K2319/00 , C07K2319/50 , C12N15/70
摘要： Disrupting the expression of endogenous Escherichia host cell genes gcvA and spr provides mutant host cells having increased heterologous peptide production. The addition of a genetic modification to the coding region of gene yejM further enhances peptide production and facilitates easier downstream processing. Recombinant Escherichia host cells are provided as well as methods of using such host cells for heterologous peptide production.
摘要翻译： 破坏内源性大肠杆菌宿主细胞基因gcvA和spr的表达提供具有增加的异源肽产生的突变宿主细胞。 在基因yejM的编码区添加遗传修饰进一步增强了肽的产生，并且便于下游处理。 提供重组埃希氏杆菌宿主细胞以及使用这种宿主细胞进行异源肽生产的方法。
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公开(公告)号：US07098000B2
公开(公告)日：2006-08-29
申请号：US10860291
申请日：2004-06-03
申请人： Qiong Cheng , Luan Tao
发明人： Qiong Cheng , Luan Tao
CPC分类号： C12N9/0083 , C12N9/1085 , C12N15/52 , C12P23/00
摘要： The present invention provides methods to engineer microorganisms for the production of C30-aldehyde carotenoids. Specifically, various combinations of crtM, sqs, crtN and crtN2 genes from Staphylococcus aureus and Methylomonas sp. 16a can be co-expressed in transformant hosts, leading to the production of diaponeurosporene monoaldehyde, diapocarotene monoaldehyde, and/or diapocarotene dialdehyde. In a preferred embodiment, the genetically engineered pathway is introduced into a strain of Escherichia coli that has been engineered for the expression of carotenoids, and the C30-carotenoid product is diapocarotene dialdehyde.
摘要翻译： 本发明提供了工程微生物用于生产C 30 N-醛类胡萝卜素的方法。 具体来说，来自金黄色葡萄球菌和Methylomonas sp。的crtM，sqs，crtN和crtN2基因的各种组合。 16a可以在转化体宿主中共表达，导致产生二头孢噻吩一醛，二卡非罗芬单醛和/或二卡必康二醛。 在优选的实施方案中，将经遗传工程改造的途径引入已被改造用于表达类胡萝卜素的大肠杆菌菌株中，并且C 30-30-类胡萝卜素产物是二卡必康二醛。
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公开(公告)号：US07064196B2
公开(公告)日：2006-06-20
申请号：US10808979
申请日：2004-03-25
申请人： Qiong Cheng , Luan Tao
发明人： Qiong Cheng , Luan Tao
CPC分类号： C12N9/001 , C12N9/00 , C12N9/0004 , C12N9/0071 , C12N9/0083 , C12N9/1051 , C12N9/1085 , C12N9/90 , C12N15/52 , C12P23/00
摘要： Genes have been isolated from strain DC260, a member of the Enterobacteriaceae family, encoding geranylgeranyl pyrophosphate (GGPP) synthetase (CrtE), phytoene synthase (CrtB), phytoene desaturase (CrtI), lycopene cyclase (CrtY), β-carotene hydroxylase (CrtZ), and zeaxanthin glucosyl transferase (CrtX) activity. The genes and their products are useful for the conversion of farnesyl pyrophosphate to carotenoids. Vectors containing those DNA segments, host cells containing the vectors and methods for producing those enzymes by recombinant DNA technology in transformed host organisms are disclosed.
摘要翻译： 基因已经从肠杆菌科家族的成员DC260中分离出来，编码香叶基香叶基焦磷酸（GGPP）合成酶（CrtE），八氢番茄红素合酶（CrtB），八氢番茄红素去饱和酶（CrtI），番茄红素环化酶（CrtY），β-胡萝卜素羟化酶 ）和玉米黄质葡萄糖基转移酶（CrtX）活性。 这些基因及其产物可用于将焦磷酸法呢基转化成类胡萝卜素。 公开了含有这些DNA片段的载体，含有载体的宿主细胞和通过重组DNA技术在转化的宿主生物体中产生这些酶的方法。
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公开(公告)号：US06929928B2
公开(公告)日：2005-08-16
申请号：US10808807
申请日：2004-03-24
申请人： Qiong Cheng , Natalia Sedkova , Luan Tao
发明人： Qiong Cheng , Natalia Sedkova , Luan Tao
IPC分类号： C07H21/04 , C12N20060101 , C12N1/14 , C12N1/20 , C12N1/21 , C12N9/00 , C12N9/02 , C12N9/10 , C12N9/90 , C12N15/52 , C12N15/82 , C12P5/00 , C12P5/02 , C12P7/26 , C12P23/00
CPC分类号： C12N9/00 , C12N9/0004 , C12N9/001 , C12N9/0083 , C12N9/1051 , C12N9/1085 , C12N9/90 , C12N15/52 , C12N15/825 , C12P23/00 , C12Y205/01029 , Y02E50/343
摘要： A unique carotenogenic biosynthetic gene cluster has been isolated from Panteoa agglomerans strain DC404, wherein the genetic organization of the cluster is crtE-idi-crtY-crtI-crtB-crtZ. The genes contained within this cluster encode geranylgeranyl pyrophosphate (GGPP) synthetase (CrtE), isopentenyl pyrophosphate isomerase (Idi), lycopene cyclase (CrtY), phytoene desaturase (CrtI), phytoene synthase (CrtB), and β-carotene hydroxylase (CrtZ). The gene cluster, genes and their products are useful for the conversion of farnesyl pyrophosphate to carotenoids. Vectors containing those DNA segments, host cells containing the vectors and methods for producing those enzymes by recombinant DNA technology in transformed host organisms are disclosed.
摘要翻译： 已经从成团Panthena菌株DC404中分离出独特的致病基因生物合成基因簇，其中簇的遗传组织为crtE-idi-crtY-crtI-crtB-crtZ。 包含在该簇内的基因编码er牛儿基焦磷酸（GGPP）合成酶（CrtE），异戊烯焦磷酸异构酶（Idi），番茄红素环化酶（CrtY），八氢番茄红素去饱和酶（CrtI），八氢番茄红素合酶（CrtB）和β-胡萝卜素羟化酶（CrtZ） 。 基因簇，基因及其产物可用于将焦磷酸法呢基转化成类胡萝卜素。 公开了含有这些DNA片段的载体，含有载体的宿主细胞和通过重组DNA技术在转化的宿主生物体中产生这些酶的方法。
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公开(公告)号：US06794165B2
公开(公告)日：2004-09-21
申请号：US10272419
申请日：2002-10-16
IPC分类号： C12P744
摘要： A gene cluster has been isolated from an Acinetobacter sp. that encodes the enzymes expected to convert cyclohexanol to adipic acid. The entire gene cluster has been cloned and all open reading frames have been sequenced. Cosmid clones have been identified containing the gene cluster. Demonstration of conversion of cyclohexanol to adipic acid has been made with the recombinant E. coli host strain containing the cosmids.
摘要翻译： 已经从不动杆菌属中分离出一个基因簇。 其编码预期将环己醇转化为己二酸的酶。 已经克隆了整个基因簇，并且所有的开放阅读框已被测序。 确定了含有基因簇的粘粒克隆。 用包含粘粒的重组大肠杆菌宿主菌已经证实了环己醇转化为己二酸。
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