公开(公告)号：US08409798B2

公开(公告)日：2013-04-02

申请号：US11547948

申请日：2005-04-08

申请人： Markus Luy ,  Matthias Rüsing

发明人： Markus Luy ,  Matthias Rüsing

IPC分类号： C12Q1/68

CPC分类号： C12Q1/689

摘要： The invention relates to a method for the rapid and simple identification of PUFA PKS in microorganisms. Said method is characterized by the fact that DNA sections representing PKS that produce specifically for PUFA (polyunsaturated fatty acids) are reproduced in vitro. The PUFA PKS-specific amino acid sequence LGIDSIKRVEIL makes it possible to derive oligonucleotides which are used for the efficient PCR screening for PUFA PKS genes or PUFA-producing microorganisms. The inventive method is particularly suitable for the high throughput screening of microorganisms for PUFA PKS genes.

摘要翻译： 本发明涉及用于快速和简单地鉴定微生物中PUFA PKS的方法。 所述方法的特征在于代表PKS产生的专门用于PUFA（多不饱和脂肪酸）的DNA片段在体外再现。 PUFA PKS特异性氨基酸序列LGIDSIKRVEIL可以获得用于有效PCR筛选PUFA PKS基因或产生PUFA的微生物的寡核苷酸。 本发明的方法特别适合用于PUFA PKS基因的微生物的高通量筛选。

公开(公告)号：US08900831B2

公开(公告)日：2014-12-02

申请号：US10578968

申请日：2004-11-10

申请人： Matthias Rüsing ,  Markus Luy

发明人： Matthias Rüsing ,  Markus Luy

IPC分类号： C12P7/64 ,  C12N1/00

CPC分类号： C12P7/6427 ,  A23K10/37 ,  A23L33/12 ,  A23V2002/00 ,  C12N1/12 ,  C12P7/6472

摘要： The invention relates to an optimized method for cultivating microorganisms of the genus Thraustochytriales, according to which the microorganisms are cultivated in a low salt medium without adding sodium salts and chloride salts, the total salt content being less than 3.5 g/L (corresponding to less than 10 percent of sea water salt content), whereupon the PUFAs are isolated from the microorganisms and/or the medium. The invention especially relates to novel optimized media having a substantially reduced total salt content, above all a particularly reduced NaCl content. The production of PUFAs can be substantially improved and significantly simplified by using a novel combination of different salts as a media composition in which the overall weight ratios of ions do not exceed 1.75 g/L. Furthermore, said medium preferably contains no added sodium salt and chloride salt at all, which helps prevent environmental damages caused by wastewaters containing salt.

摘要翻译： 本发明涉及一种用于培养破囊壶菌属微生物的优化方法，根据该方法，微生物在低盐培养基中培养而不添加钠盐和氯化物盐，总盐含量小于3.5g / L（相当于少于 超过10％的海水盐含量），由此从微生物和/或介质中分离PUFA。 本发明特别涉及具有显着降低的总盐含量，特别是特别降低的NaCl含量的新型优化的介质。 通过使用不同盐的新型组合作为其中离子的总重量比不超过1.75g / L的介质组合物，可以显着改善和显着地简化PUFA的生产。 此外，所述介质优选根本不含有添加的钠盐和氯化物盐，这有助于防止由含有盐的废水引起的环境损害。

公开(公告)号：US08889382B2

公开(公告)日：2014-11-18

申请号：US10578965

申请日：2004-11-10

申请人： Markus Luy ,  Matthias Rüsing

发明人： Markus Luy ,  Matthias Rüsing

IPC分类号： C12P7/64 ,  C12N1/00

CPC分类号： C12P7/6427 ,  C12N1/14 ,  C12P7/6472

摘要： The invention relates to an optimized method for the production of PUFAs by cultivating microorganisms belonging to the group of Stramenopiles in a fermentation medium that is pH-stabilized using calcium carbonate and comprises 3-15 g/L CaCO3, whereupon the PUFAs are isolated from the microorganisms and/or the medium. The invention particularly relates to novel optimized media having a different CaCO3 content. By using adequate quantities of CaCO3, the process can be significantly simplified during fermentation while greater quantities of DHA can be obtained at an increased oil content in the biomass. They allow microorganisms belonging to the Stramenopiles to be fermented without controlling the pH, thereby substantially improving and significantly simplifying PUFA production.

摘要翻译： 本发明涉及通过在使用碳酸钙进行pH稳定化的发酵培养基中培养属于Stramenopiles组的微生物并且包含3-15g / L CaCO 3来生产PUFA的优化方法，由此将PUFA从 微生物和/或培养基。 本发明特别涉及具有不同CaCO3含量的新型优化介质。 通过使用足够量的CaCO 3，可以在发酵期间显着简化该过程，同时在生物质中增加的油含量可获得更多量的DHA。 它们允许属于Stramenopiles的微生物发酵而不控制pH值，从而显着改善和显着简化PUFA生产。

公开(公告)号：US07939305B2

公开(公告)日：2011-05-10

申请号：US11547921

申请日：2005-04-08

申请人： Markus Luy ,  Matthias Rüsing ,  Thomas Kiy

发明人： Markus Luy ,  Matthias Rüsing ,  Thomas Kiy

IPC分类号： C12N9/00

CPC分类号： C12P7/6472 ,  C12N15/52

摘要： The invention relates to genes which are coded for sequences specific to polyketide synthases (PKS). The thus synthesized PKS is characterized by the enzymatic capacity thereof to produce PUFAs (polyunsaturated fatty acids). The invention also relates to the identification of the corresponding DNA-sequences, in addition to the use of said nucleotide sequences for the production of recombined and/or transgenic organisms.

摘要翻译： 本发明涉及编码针对聚酮化合物合酶（PKS）特异性序列的基因。 由此合成的PKS的特征在于其产生PUFA（多不饱和脂肪酸）的酶能力。 除了使用所述核苷酸序列产生重组和/或转基因生物外，本发明还涉及鉴定相应的DNA序列。

公开(公告)号：US07105324B2

公开(公告)日：2006-09-12

申请号：US10457452

申请日：2003-06-10

申请人： Matthias Rüsing ,  Thomas Schweins ,  Petra Dresler ,  Wolfgang Stock ,  Thomas Kiy

发明人： Matthias Rüsing ,  Thomas Schweins ,  Petra Dresler ,  Wolfgang Stock ,  Thomas Kiy

IPC分类号： C12P7/64

CPC分类号： C12N9/90 ,  C07K2319/00

摘要： The present invention relates to nucleic acids isolated from Tetrahymena which code for a ciliate-specific triterpenoid cyclase. The inventive nucleotide sequences and the polypeptide sequences derived therefrom demonstrate a surprisingly minimal sequence identity to known isoprenoid cyclases. The invention also relates to the use of nucleic acids for the regulation of triterpenoid cyclase expression in a host organism, as well as the targeted knockout or repriming of the triterpenoid cyclase gene. As a result of the altered expression of the triterpenoid cyclase, it is possible to modify and enrich the levels of multiple unsaturated fatty acids in the host organism.

公开(公告)号：US06962800B2

公开(公告)日：2005-11-08

申请号：US10395433

申请日：2003-03-24

申请人： Thomas Kiy ,  Matthias Rüsing

发明人： Thomas Kiy ,  Matthias Rüsing

IPC分类号： C12N15/09 ,  C07K14/765 ,  C12N1/11 ,  C12N5/06 ,  C12N15/79 ,  C12P21/02 ,  C12P21/06

CPC分类号： C07K14/765 ,  C12N15/79

摘要： The present invention concerns a method for production of recombinant human proteins, in which Tetrahymena cells are transformed with recombinant DNA containing at least one functional gene that codes for a human protein, the recombinant Tetrahymena cells are cultured, in which the gene that codes for a human protein is expressed and the proteins are then isolated. The present invention also concerns a corresponding method, in which the gene that codes for a human protein contains a human leader sequence that leads to secretion of the expressed protein.

摘要翻译： 本发明涉及重组人蛋白质的制备方法，其中用包含编码人蛋白质的至少一个功能基因的重组DNA转化四膜虫细胞，培养重组四膜虫细胞，其中编码 表达人蛋白质，然后分离蛋白质。 本发明还涉及一种相应的方法，其中编码人蛋白质的基因含有导致表达的蛋白质分泌的人前导序列。

公开(公告)号：US20120172576A1

公开(公告)日：2012-07-05

申请号：US13339454

申请日：2011-12-29

申请人： Matthias Rüsing

发明人： Matthias Rüsing

IPC分类号： C07K14/00 ,  C12N1/11 ,  C12P21/06 ,  C12N15/74

CPC分类号： C12N9/0083 ,  C12N9/90 ,  C12N15/79 ,  C12Y114/19001 ,  C12Y114/19003

摘要： The present invention concerns a method for production of genetically modified (recombinant) protists without using negative selection markers, in which an auxotrophic mutant of the protist is produced, this mutant is then transformed with recombinant DNA containing at least one gene for complementation of the corresponding auxotrophy, and the resulting recombinant protist is finally selected on a minimal medium that makes possible growth of only the correspondingly complemented protist. The present invention also concerns an efficient method for production of proteins by protists so modified, in which the gene for the protein being produced is coupled to the marker gene.

摘要翻译： 本发明涉及一种生产基因修饰（重组）原生生物的方法，而不使用产生原生生物的营养缺陷突变体的阴性选择标记，然后用含有至少一个基因的重组DNA转化该突变体，以补充相应的 营养缺陷型，并且最终在最小的培养基上选择所得的重组原生生物，这使得只有相应补充的原生生物可能生长。 本发明还涉及通过如此修饰的原生质体生产蛋白质的有效方法，其中所产生的蛋白质的基因与标记基因偶联。

公开(公告)号：US06645751B2

公开(公告)日：2003-11-11

申请号：US09725735

申请日：2000-11-30

申请人： Matthias Rüsing ,  Thomas Schweins ,  Petra Dresler ,  Wolfgang Stock ,  Thomas Kiy

发明人： Matthias Rüsing ,  Thomas Schweins ,  Petra Dresler ,  Wolfgang Stock ,  Thomas Kiy

IPC分类号： C12N990

CPC分类号： C12N9/90 ,  C07K2319/00

摘要： The present invention relates to nucleic acids isolated from Tetrahymena which code for a ciliate-specific triterpenoid cyclase. The inventive nucleotide sequences and the polypeptide sequences derived therefrom demonstrate a surprisingly minimal sequence identity to known isoprenoid cyclases. The invention also relates to the use of nucleic acids for the regulation of triterpenoid cyclase expression in a host organism, as well as the targeted knockout or repriming of the triterpenoid cyclase gene. As a result of the altered expression of the triterpenoid cyclase, it is possible to modify and enrich the levels of multiple unsaturated fatty acids in the host organism.

摘要翻译： 本发明涉及从四膜虫分离的编码细胞特异性三萜环化酶的核酸。 本发明的核苷酸序列及其衍生的多肽序列与已知的类异戊二烯环化酶表现出惊人的最小序列同一性。 本发明还涉及核酸用于调节宿主生物体中三萜环化酶表达的用途，以及三萜环化酶基因的靶向敲除或重新引导。 作为三萜环化酶表达改变的结果，有可能修饰和丰富宿主生物体中多种不饱和脂肪酸的含量。