Alleles of the rel gene of coryneform bacteria 机翻标题: 暂无翻译,请尝试点击翻译按钮。

公开号/公开日
EP2061881 B1 2015-09-09 [EP2061881]EP2061881 A2 2009-05-27 [EP2061881] / 2015-09-092009-05-27
申请号/申请日
2007EP-0821334 / 2007-10-15
发明人
BATHE BRIGITTE;KREUTZER CAROLINE;THIERBACH GEORG;
申请人
EVONIK DEGUSSA;
主分类号
IPC分类号
A23L-001/227C12N-001/21C12N-009/12C12N-015/54C12N-015/63C12N-015/69C12P-013/04C12P-013/08C12R-001/15
摘要
(EP2061881) An isolated mutant of a coryneform bacterium comprising a gene coding for a polypeptide having GTP-pyrophosphate kinase activity, wherein said polypeptide comprises an amino acid sequence in which one of the proteinogenic amino acids other than L-proline is present in position 38 or a corresponding or comparable position.  In addition, an isolated polynucleotide encoding a polypeptide having GTP-pyrophosphate kinase enzyme activity, a vector comprising the isolated polynucleotide, a recombinant microorganism comprising the vector, and a process for preparing the recombinant coryneform bacterium is described.  A method for over-expressing a GTP-pyrophosphate kinase, a method of preparing an L-amino acid, an L-lysine comprising and L-tryptophan comprising feed is also described.  (From US8293514 B2)
机翻摘要
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地址
代理人
代理机构
;
优先权号
2006DE-10048882 2006-10-17 2007WO-EP60967 2007-10-15
主权利要求
(EP2061881) 1. Isolated mutants of coryneform bacteria, which comprise a gene coding for a polypeptide having GTP-pyrophosphate kinase activity, characterized in that said polypeptide comprises an amino acid sequence selected from the group consisting of a) amino acid sequence according to SEQ ID NO: 6, and b) amino acid sequence according to SEQ ID NO: 6 including one or more, to a maximum of 5, insertions or deletions of amino acids, which comprises L-leucine in position 38 or in the position of the amino acid sequence that corresponds to position 38, wherein a position that corresponds to position 38 is to be understood as meaning that insertion or deletion of a codon coding for an amino acid in the N-terminal region based on position 38 of SEQ ID NO: 6 of the encoded polypeptide formally increases, in the case of an insertion, or decreases, in the case of a deletion, the indicated position and indicated length, in each case by one unit,  with L-glutamic acid optionally being present in position 262. 2. Mutants of coryneform bacteria according to Claim 1, characterized in that the coryneform bacteria are a bacterium selected from the group consisting of Corynebacterium efficiens, Corynebacterium glutamicum, Corynebacterium thermoaminogenes and Corynebacterium aminogenes. 3. Mutants of coryneform bacteria according to Claim 1 or 2, characterized in that they are L-amino acid-secreting bacteria, preferably L-lysine-, L-valine-, L-isoleucine-, L-tryptophan- or L-homoserine-secreting bacteria. 4. Mutants according to one of Claims 1 to 3, characterized in that the gene comprises the nucleotide sequence of SEQ ID NO:5 or SEQ ID NO:7, the guanine in position 785 of SEQ ID NO:5 or the guanine in position 1535 of SEQ ID NO:7 optionally being replaced by adenine. 5. Isolated polynucleotide, characterized in that the encoded protein comprises an amino acid sequence selected from the group consisting of a) an amino acid sequence according to SEQ ID NO:6, and b) an amino acid sequence according to SEQ ID NO:6, including one or more, to a maximum of 5, amino acid insertions or deletions, which comprises L-leucine in position 38 or in the position of the amino acid sequence that corresponds to position 38, wherein a position that corresponds to position 38 is to be understood as meaning that insertion or deletion of a codon coding for an amino acid in the N-terminal region based on position 38 of SEQ ID NO:6 of the encoded polypeptide formally increases, in the case of an insertion, or decreases, in the case of a deletion, the indicated position and indicated length, in each case by one unit, L-glutamic acid being optionally present in position 262. 6. Isolated polynucleotide according to Claim 5, characterized in that the nucleotide sequence thereof comprises SEQ ID NO:5, adenine optionally being present in position 785. 7. Recombinant microorganism comprising the isolated polynucleotide according to Claim 5 or 6,  which is preferably a coryneform bacterium or a bacterium of the genus Escherichia,  particularly preferably the genus Corynebacterium, very particularly preferably the species Corynebacterium glutamicum. 8. Vector comprising the isolated polynucleotide according to Claim 5 or 6. 9. Recombinant microorganism comprising the vector according to Claim 8,  preferably a recombinant microorganism which is a coryneform bacterium or a bacterium of the genus Escherichia,  particularly preferably the genus Corynebacterium, very particularly preferably the species Corynebacterium glutamicum. 10. Process for preparing an L-amino acid, which comprises a) fermenting an isolated coryneform bacterium in a suitable medium, said bacterium harboring at least one copy of a gene encoding a polypeptide having GTP-pyrophosphate kinase enzyme activity, wherein the polypeptide comprises the amino acid sequence according to SEQ ID NO: 6, or the amino acid sequence according to SEQ ID NO: 6 including one to a maximum of 5 insertions or deletions, with L-leucine being present in position 38 or in the position of the amino acid sequence that corresponds to position 38, and wherein a position that corresponds to position 38 is to be understood as meaning that insertion or deletion of a codon coding for an amino acid in the N-terminal region based on position 38 of SEQ ID NO:6 of the encoded polypeptide formally increases, in the case of an insertion, or decreases, in the case of a deletion, the indicated position and indicated length, in each case by one unit, and b) concentrating the L-amino acid in the fermentation broth or in the cells of said bacterium. 11. Process according to Claim 10, characterized in that the isolated coryneform bacterium is a mutant according to Claims 1 to 4, or a recombinant coryneform bacterium which harbors an isolated polynucleotide according to Claim 5 or 6 or which has been prepared using the latter. 12. Process according to Claim 10 or 11, characterized in that the L-amino acid is isolated or collected and preferably purified or  in that the L-amino acid is isolated or collected together with components of the fermentation broth and/or the biomass (> 0 to 100%). 13. Process according to one of Claims 10 to 12, characterized in that it comprises a) removing an amount of from 0 to 100% of the biomass produced from the fermentation broth obtained in step b) of Claim 12, and b) preparing from the broth obtained in step a) an essentially dry and shaped product by a method selected from the group consisting of granulation, compacting, spray drying and extrusion. 14. Process according to Claim 13, characterized in that, in the case of the amino acid L-lysine, an acid selected from the group consisting of sulfuric acid, hydrochloric acid and phosphoric acid is added to the fermentation broth prior to or after step a) or  in that water is removed from the broth obtained prior to or after step a) and/or in that the shaped product obtained in or during step b) is sprayed with an oil or  in that the following steps are carried out a) filtering the fermentation broth, preferably by using a membrane filter, so as to obtain a biomass-containing sludge and a filtrate, b) concentrating said filtrate, preferably so as to obtain a solids content of from 48 to 52% by weight, c) granulating the concentrate obtained in step b), preferably at a temperature of from 50 deg.C to 62 deg.C, and d) coating the granules obtained in c) with one or more of the coating agents.
法律状态
(EP2061881) LEGAL DETAILS FOR EP2061881  Actual or expected expiration date=2027-10-15    Legal state=ALIVE    Status=GRANTED     Event publication date=2007-10-15  Event code=EP/APP  Event indicator=Pos  Event type=Examination events  Application details  Application country=EP EP07821334  Application date=2007-10-15  Standardized application number=2007EP-0821334     Event publication date=2009-05-27  Event code=EP/A2  Event type=Examination events  Application published without search report  Publication country=EP  Publication number=EP2061881  Publication stage Code=A2  Publication date=2009-05-27  Standardized publication number=EP2061881     Event publication date=2009-05-27  Event code=EP/17P  Event indicator=Pos  Event type=Examination events  Request for examination filed Pruefungsantrag gestellt  Effective date of the event=2009-03-17     Event publication date=2009-05-27  Event code=EP/AK  Event indicator=Pos  Event type=Designated states  Designated contracting states: Benannte vertragsstaaten AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IS IT LI LT LU LV MC MT NL PL PT RO SE SI SK TR    Event publication date=2009-05-27  Event code=EP/AX  Event indicator=Pos  Event type=Designated states  Extension of the European patent to Request for extension of the european patent to AL BA HR MK RS    Event publication date=2009-12-02  Event code=EP/17Q  Event indicator=Pos  Event type=Examination events  First examination report Erster pruefungsbescheid  Effective date of the event=2009-11-02     Event publication date=2010-01-20  Event code=EP/DAX  Event indicator=Neg  Event type=Designated states  Request for extension of the european patent (to any country) deleted    Event publication date=2015-06-24  Event code=EP/INTG  Event indicator=Pos  Event type=Examination events  Announcement of intention to grant  Effective date of the event=2015-06-01     Event publication date=2015-09-09  Event code=EP/B1  Event indicator=Pos  Event type=Event indicating In Force  Patent specification  Publication country=EP  Publication number=EP2061881  Publication stage Code=B1  Publication date=2015-09-09  Standardized publication number=EP2061881     Event publication date=2015-09-09  Event code=EP/AK  Event indicator=Pos  Event type=Designated states  Designated contracting states: Benannte vertragsstaaten AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IS IT LI LT LU LV MC MT NL PL PT RO SE SI SK TR    Event publication date=2016-08-17  Event code=EP/26N  Event indicator=Pos  Event type=No opposition filed  No opposition filed Kein einspruch eingelegt  Effective date of the event=2016-06-10  MEMBER STATE LEGAL DETAILS FOR AT  Actual or expected expiration date=2015-10-15    Legal state=DEAD    Status=LAPSED   Corresponding cc:  Designated or member state=AT Corresponding appl: AT748152    Event publication date=2015-09-15  Event code=EP/REG  Event code=AT/REF  Event indicator=Pos  Event type=Entry into national phase  Reference to a national code Reference to at number (ep patent enters austrian national phase) Nennung der e-nummer (ep patent tritt in at nationale phase ein) Corresponding cc:  Designated or member state=AT   Effective date of the event=2015-09-15     Event publication date=2016-12-15  Event code=EP/REG  Event code=AT/MM01  Event indicator=Neg  Event type=Event indicating Not In Force  Reference to a national code Lapse because of not paying annual fees Loeschung wegen nichtzahlung der jaehrlichen gebuehr Corresponding cc:  Designated or member state=AT   Effective date of the event=2015-10-15     Event publication date=2017-02-28  Event code=EP/PG25  Event indicator=Neg  Event type=Event indicating Not In Force  Event type=Payment or non-payment notifications  Lapsed in a contracting state announced via postgrant inform. from nat. office to EPO LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES  Corresponding cc:  Designated or member state=AT  MEMBER STATE LEGAL DETAILS FOR CH  Actual or expected expiration 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type=Payment or non-payment notifications  Lapsed in a contracting state announced via postgrant inform. from nat. office to EPO LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT  Corresponding cc:  Designated or member state=FI  MEMBER STATE LEGAL DETAILS FOR FR  Actual or expected expiration date=2027-10-15    Legal state=ALIVE    Status=GRANTED   Corresponding cc:  Designated or member state=FR     Event publication date=2015-10-23  Event code=EP/REG  Event code=FR/PLFP  Event indicator=Pos  Event type=Payment or non-payment notifications  Reference to a national code Fee payment Derniere annuite payee   Annual fees payment date=2015-10-23     Year of payment of annual fees=9  Corresponding cc:  Designated or member state=FR     Event publication date=2016-10-20  Event code=EP/REG  Event code=FR/PLFP  Event indicator=Pos  Event type=Payment or non-payment notifications  Reference to a national code Fee payment 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office to EPO LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT  Corresponding cc:  Designated or member state=GR  MEMBER STATE LEGAL DETAILS FOR IE  Actual or expected expiration date=2016-07-27    Legal state=DEAD    Status=LAPSED   Corresponding cc:  Designated or member state=IE     Event publication date=2015-10-07  Event code=EP/REG  Event code=IE/FG4D  Event indicator=Pos  Event type=Event indicating In Force  Reference to a national code European patents granted designating ireland Corresponding cc:  Designated or member state=IE  LANGUAGE OF EP DOCUMENT: GERMAN     Event publication date=2016-07-27  Event code=EP/REG  Event code=IE/MM4A  Event indicator=Neg  Event type=Event indicating Not In Force  Reference to a national code Patent lapsed Corresponding cc:  Designated or member state=IE     Event publication date=2016-10-31  Event code=EP/PG25  Event indicator=Neg  Event type=Event indicating Not In Force  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专利类型码
B1A2
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