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    • 1. 发明申请
    • BI-DIRECTIONAL SELECTION MARKERS WITH IMPROVED ACTIVITY
    • 具有改进活动的双向选择标记
    • WO2007118836A1
    • 2007-10-25
    • PCT/EP2007/053549
    • 2007-04-12
    • DSM IP ASSETS B.V.BERG, VAN DEN, Marco AlexanderBOVENBERG, Roelof, Ary, Lans
    • BERG, VAN DEN, Marco AlexanderBOVENBERG, Roelof, Ary, Lans
    • C12N15/55C12N9/80
    • C12N9/80
    • The present invention discloses a polypeptide selected from the group consisting of: a polypeptide having an amino acid sequence according to SEQ ID NO 3, a polypeptide having an amino acid sequence according to SEQ ID NO 6, a polypeptide having an amino acid that is substantially homologous to the sequence of SEQ ID NO 3 and a polypeptide having an amino acid that is substantially homologous to the sequence of SEQ ID NO 6, the polypeptide displaying acetamidase activity and providing a reverse selection on fluoroacetamide with an efficiency of at least 95%, preferably at least 96%, more preferably at least 97%, more preferably at least 98%, more preferably at least 99%, most preferably 100%. The gene encoding the polypeptide of the invention is used as an efficient bi-directional selection marker in the construction of selection marker free strains, in particular for processes for the production of a compound of interest.
    • 本发明公开了一种选自以下的多肽:具有根据SEQ ID NO 3的氨基酸序列的多肽,具有根据SEQ ID NO 6的氨基酸序列的多肽,具有基本上 与SEQ ID NO 3的序列同源,以及具有与SEQ ID NO 6的序列基本同源的氨基酸的多肽,所述多肽显示乙酰胺酶活性,并提供效率至少为95%的氟代乙酰胺的反向选择, 优选至少96%,更优选至少97%,更优选至少98%,更优选至少99%,最优选100%。 编码本发明多肽的基因在构建选择标记免疫菌株时被用作有效的双向选择标记,特别是用于生产感兴趣的化合物的方法。
    • 10. 发明申请
    • IMPROVED IN VIVO PRODUCTION OF CEPHALOSPORINS
    • 改善头孢菌素的体内生产
    • WO0037671A3
    • 2000-09-14
    • PCT/EP9910292
    • 1999-12-21
    • DSM NVBOVENBERG ROELOF ARY LANSKERKMAN RICHARDKOENHEN ERIC
    • BOVENBERG ROELOF ARY LANSKERKMAN RICHARDKOENHEN ERIC
    • C12P35/06C12N9/10C12P35/00C12R1/82
    • C12P35/00C12N9/1029
    • The present invention discloses a process for the production of 7-amino cephalosporanic acid (7-ACA) or a derivative thereof comprising the steps of fermenting a P. chrysogenum strain being transformed with an expression construct comprising a nucleotide sequence encoding expandase, hydroxylase and acetyltransferase activity in the presence of a suitable acyl side chain precursor, or a salt or ester thereof, such that an N-acylated 7-ACA compound is produced, N-deacylating the thus produced N-acylated 7-ACA compound and, optionally, acylating the free amino group and/or substituting the 3' acetate group with a side chain suitable to form a cephalosporin antibiotic, characterised in that the nucleotide sequence encoding the acetyltransferase is derived from Acremonium chrysogenum and starts at the second ATG of the open reading frame as present in said nucleotide sequence.
    • 本发明公开了生产7-氨基头孢菌酸(7-ACA)或其衍生物的方法,其包括发酵产黄青霉菌菌株的步骤,所述产黄青霉菌菌株用包含编码扩环酶,羟化酶和乙酰​​转移酶的核苷酸序列的表达构建体转化 在合适的酰基侧链前体或其盐或酯存在下的活性,从而产生N-酰化的7-ACA化合物,将由此产生的N-酰化的7-ACA化合物N-脱酰基化,和任选地酰化 游离氨基和/或用适于形成头孢菌素抗生素的侧链取代3'乙酸酯基团,其特征在于编码乙酰转移酶的核苷酸序列来自Acremonium chrysogenum并开始于开放阅读框的第二个ATG, 存在于所述核苷酸序列中。