公开(公告)号：US06558921B1

公开(公告)日：2003-05-06

申请号：US09631022

申请日：2000-08-02

申请人： Jose Luis Barredo Fuente ,  Marta Rodriguez Saiz ,  Alfonso J. Collados de la Vieja ,  Migeul Angel Moreno Valle ,  Francisco Salto Maldonado ,  Bruno Diez Garcia

发明人： Jose Luis Barredo Fuente ,  Marta Rodriguez Saiz ,  Alfonso J. Collados de la Vieja ,  Migeul Angel Moreno Valle ,  Francisco Salto Maldonado ,  Bruno Diez Garcia

IPC分类号： C12P102

CPC分类号： C12N15/1137 ,  C07K2319/00 ,  C07K2319/02 ,  C07K2319/036 ,  C12N9/0016 ,  C12N9/2402 ,  C12N15/625 ,  C12N15/70 ,  C12N15/80 ,  C12Y104/01002 ,  C12Y302/01052

摘要： An isolated DNA having the promoter sequence of the hex gene of P. chrysogenum or a DNA fragment that is hybridizable to the complement of the promoter sequence under stringent conditions and is capable of directing expression of DNA downstream of the fragment in P. chrysogenum. Also a process for promoting expression of a coding sequence of interest in a microorganism using the isolated DNA and a process to block expression of a gene of interest in a microorganism using the isolated DNA are disclosed.

摘要翻译： 一种分离的DNA，其具有产黄青霉六方基因的启动子序列或在严格条件下可与启动子序列的互补体杂交的DNA片段，并且能够引导产生产黄青霉的片段下游的DNA的表达。 还公开了使用分离的DNA促进微生物中感兴趣的编码序列的表达的方法和使用分离的DNA阻断微生物中目的基因表达的过程。

公开(公告)号：US06300095B1

公开(公告)日：2001-10-09

申请号：US09171337

申请日：1999-05-14

申请人： Jose Luis Barredo Fuente ,  Marta Rodriguez Saiz ,  Alfonso J. Collados De La Vieja ,  Migeul Angel Moreno Valle ,  Francisco Salto Maldonado ,  Bruno Diez Garcia

发明人： Jose Luis Barredo Fuente ,  Marta Rodriguez Saiz ,  Alfonso J. Collados De La Vieja ,  Migeul Angel Moreno Valle ,  Francisco Salto Maldonado ,  Bruno Diez Garcia

IPC分类号： C07K14385

CPC分类号： C12N15/1137 ,  C07K2319/00 ,  C07K2319/02 ,  C07K2319/036 ,  C12N9/0016 ,  C12N9/2402 ,  C12N15/625 ,  C12N15/70 ,  C12N15/80 ,  C12Y104/01002 ,  C12Y302/01052

摘要： The invention relates to promoters of the genes glutamate dehydrogenase, &bgr;-acetylhexosaminidase and &ggr;-actin and their use in systems of expression, secretion and anti-sense of filamentary fungi. The invention also relates to the use of the promoters of the genes which code: (I) glutamate dehydrogenase NADP depending (EC.1.4.1.4) of Penicillium chrysogenum, (II) &ggr;-N-actylhexosaminidase (EC.3.2.1.52) of Penicillium chrysogenum and (III) &ggr;-actin of Penicillium chrysogenum and Acrimonium chrysogenum, which can be used for the construction of potent vectors of expression and secretion useful both for P. chrysogenum and for A. chrysogenum and related species. These promoters can also be used for blocking the genic expression through anti-sense construction. Under the control of the above mentioned promoters, it is possible to conduct the expression of other genes in filamentary fungi, thereby increasing the production of antibiotics and/or proteins inherent to the same.

摘要翻译： 本发明涉及谷氨酸脱氢酶，β-乙酰氨基己糖苷酶和γ-肌动蛋白基因的启动子及其在丝状真菌表达，分泌和反义系统中的用途。 本发明还涉及编码：（I）谷氨酸脱氢酶NADP依赖于产生产青青霉（EC.1.4.1.4），（II）γ-N-酰基六氨基甲酰氨基（EC.3.2.1.52）的基因的启动子的使用， 产黄青霉和（III）产黄青霉菌和产黄青霉素的γ-肌动蛋白，可用于构建有效的表达和分泌载体，可用于产黄青素和产黄青霉及相关物种。 这些启动子也可用于通过反义结构阻断基因表达。 在上述启动子的控制下，可以在丝状真菌中进行其他基因的表达，从而增加其固有的抗生素和/或蛋白质的产生。

公开(公告)号：US06187574B1

公开(公告)日：2001-02-13

申请号：US08973914

申请日：1998-04-28

申请人： Jose Luis Garcia Lopez ,  Estrella Cortes Rubio ,  Jorge Alonso Palacios ,  Jose Luis Barredo Fuente ,  Bruno Diez Garcia ,  Miguel Angel Moreno Valle ,  Carmen Schleissner Sanchez ,  Alfonso Collados de la Vieja ,  Alejandro Vitaller Alba ,  Francisco Salto Maldonado

发明人： Jose Luis Garcia Lopez ,  Estrella Cortes Rubio ,  Jorge Alonso Palacios ,  Jose Luis Barredo Fuente ,  Bruno Diez Garcia ,  Miguel Angel Moreno Valle ,  Carmen Schleissner Sanchez ,  Alfonso Collados de la Vieja ,  Alejandro Vitaller Alba ,  Francisco Salto Maldonado

IPC分类号： C12N902

CPC分类号： C12N9/0024

摘要： Process for producing the enzyme D-amino acid oxydase of Rhodotorula gracilis in host cells. The process for the expression of the enzyme comprises isolating the complementary DNA corresponding to the messager RNA of the gene which codes for the D-amino acid oxydase of any strain of Rhodotorula gracillis producing said enzyme; fusing the fragment of DNA which codes for D-amino acid oxydase of Rhodotorula gracillis with a DNA sequence which contains a site of union to the ribosome and a high efficiency promoter sequence for the express of genes in host cells; inserting the DNA fragment into a plasmid appropriate for the host cell; cultivating the host cells transformed with said plasmid; and collecting the enzyme.

摘要翻译： 在宿主细胞中生产细小红球菌的D-氨基酸氧化酶酶的方法。 表达酶的过程包括分离与编码产生所述酶的任何红曲霉菌株的D-氨基酸氧化酶的基因的信使RNA相对应的互补DNA; 将编码红景天细菌的D-氨基酸氧化酶的DNA片段与包含与核糖体结合的位点的DNA序列和用于在宿主细胞中表达基因的高效启动子序列融合; 将DNA片段插入适合宿主细胞的质粒中; 培养用所述质粒转化的宿主细胞; 并收集酶。

公开(公告)号：US06251655B1

公开(公告)日：2001-06-26

申请号：US08952457

申请日：1998-10-28

申请人： Baltasar Minambres Rodriguez ,  Honorina Martinez Blanco ,  Elias Rodriguez Olivera ,  Belen Garcia Alonso ,  Jose Manuel Fernandez Canon ,  Jose Luis Barredo Fuente ,  Bruno Diez Garcia ,  Carmen Schleissner Sanchez ,  Miguel Angel Moreno Valle ,  Francisco Salto Maldonado ,  Jose Maria Luengo Rodriguez

发明人： Baltasar Minambres Rodriguez ,  Honorina Martinez Blanco ,  Elias Rodriguez Olivera ,  Belen Garcia Alonso ,  Jose Manuel Fernandez Canon ,  Jose Luis Barredo Fuente ,  Bruno Diez Garcia ,  Carmen Schleissner Sanchez ,  Miguel Angel Moreno Valle ,  Francisco Salto Maldonado ,  Jose Maria Luengo Rodriguez

IPC分类号： C12N120

CPC分类号： C12N9/93 ,  C12N2310/111 ,  C12P37/02

摘要： An isolated DNA encoding phenyl acetyl-CoA-ligase and a process of increasing the production of penicillin G in a strain of Penicillium chrysogenum by transforming the strain with the isolated DNA. Also vectors and host organisms having the isolated DNA.

摘要翻译： 编码苯乙酰辅酶A连接酶的分离DNA和通过用分离的DNA转化菌株来增加产黄青霉菌株中青霉素G的产生的方法。 还有具有分离的DNA的载体和宿主生物体。

公开(公告)号：US6146871A

公开(公告)日：2000-11-14

申请号：US981321

申请日：1998-08-13

申请人： Jose Luis Garcia Lopez ,  Estrella Cortes Rubio ,  Jose Manuel Guisan Seijas ,  Jose Luis Barredo Fuente ,  Bruno Diez Garcia ,  Alfonso Collados de la Vieja ,  Alejandro Vitaller Alba ,  Francisco Salto Maldonado

发明人： Jose Luis Garcia Lopez ,  Estrella Cortes Rubio ,  Jose Manuel Guisan Seijas ,  Jose Luis Barredo Fuente ,  Bruno Diez Garcia ,  Alfonso Collados de la Vieja ,  Alejandro Vitaller Alba ,  Francisco Salto Maldonado

IPC分类号： C12N15/09 ,  C12N1/21 ,  C12N9/78 ,  C12N9/84 ,  C12N9/86 ,  C12N15/00 ,  C12N15/55 ,  C12P17/14 ,  C12R1/01 ,  C12R1/19 ,  C12N9/18 ,  C12N15/70

CPC分类号： C12N9/86 ,  C12N9/84 ,  C07K2319/00

摘要： A procedure for producing a modified 7.beta.-(4-carboxybutanamide) cephalosporinase enzyme which can be purified in a single chromatographic step. The procedure for production of the enzyme involves: mutagenizing the gene which codes for the enzyme from Acinetobacter sp. ATCC 53891 by inserting a nucleotide sequence coding for six histidine residues; fusing the mutant gene with high-efficiency promoter DNA sequences; transforming Escherichia coli cells with the fusion gene construct; growing the transformed Escherichia coli cells; and recovering the enzyme by the use of supports which contain metal chelates. 7-Aminocephalosporanic acid is an important intermediate for the manufacture of a wide range of antibacterial agents of the cephalosporin family.

摘要翻译： PCT No.PCT / ES97 / 00098 Sec。 371日期1998年8月13日 102（e）日期1998年8月13日PCT 1997年4月18日PCT PCT。 公开号WO97 / 40175 PCT 日期1997年10月30日制备修饰的7β-（4-羧基丁酰胺）头孢菌素酶的方法，其可以在单个色谱步骤中纯化。 酶的制备方法包括：从不动杆菌属（Acinetobacter sp。）中诱变编码酶的基因。 ATCC 53891通过插入编码六个组氨酸残基的核苷酸序列; 将突变基因与高效启动子DNA序列融合; 用融合基因构建体转化大肠杆菌细胞; 生长转化的大肠杆菌细胞; 并通过使用含有金属螯合物的载体回收酶。 7-氨基头孢烷酸是制造头孢菌素类的广泛抗菌剂的重要中间体。

公开(公告)号：US06815189B1

公开(公告)日：2004-11-09

申请号：US09958413

申请日：2002-03-01

申请人： Javier Velasco Alvarez ,  Santiago Gutierrez Martin ,  Francisco Javier Casqueiro Blanco ,  Sonia Campoy Garcia ,  Francisco Fierro Fierro ,  Jose Luis Barredo Fuente ,  Bruno Diez Garcia ,  Juan Francisco Martin Martin

发明人： Javier Velasco Alvarez ,  Santiago Gutierrez Martin ,  Francisco Javier Casqueiro Blanco ,  Sonia Campoy Garcia ,  Francisco Fierro Fierro ,  Jose Luis Barredo Fuente ,  Bruno Diez Garcia ,  Juan Francisco Martin Martin

IPC分类号： C12N914

CPC分类号： C12N9/18 ,  C12N9/58 ,  C12P35/00 ,  C12P35/02 ,  C12P35/06

摘要： An isolated DNA sequence that encodes a peptide with CPC-acetylhydrolase enzymatic activity is disclosed. Also, a method of expressing CPC-acetylhydrolase activity that includes the steps of (a) providing a microorganism that is susceptible to transformation with the isolated DNA sequence and that, upon transformation, expresses the CPC-acetylhydrolase activity encoded by the sequence, and (b) transforming the microorganism with the DNA sequence to cause expression of the CPC-acetylhydrolase activity in the microorganism is disclosed. Also, a method for producing a microorganism with increased capacity to aid in the production of cephalosporin, which method includes the steps of (a) providing a microorganism that has CPC-AH activity by virtue of expression of the DNA sequence or of a fragment thereof coding for the CPC-acetylhydrolase enzyme activity; and (b) inactivating the activity by disrupting expression of the DNA sequence is disclosed.

摘要翻译： 公开了编码具有CPC-乙酰水解酶酶活性的肽的分离的DNA序列。 此外，表达CPC-乙酰水解酶活性的方法包括以下步骤：（a）提供易于用分离的DNA序列转化的微生物，并且在转化时表达由序列编码的CPC-乙酰水解酶活性，和 b）公开了用DNA序列转化微生物以引起微生物中的CPC-乙酰水解酶活性的表达。 另外，生产具有增加能力以帮助头孢菌素生产的微生物的方法，该方法包括以下步骤：（a）通过DNA序列或其片段的表达提供具有CPC-AH活性的微生物 编码CPC-乙酰水解酶活性; 并且（b）通过破坏DNA序列的表达而使活性失活。