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1.发明申请DNA SEQUENCE ENCODING PENICILLIN ACYLASE, NOVEL RECOMBINANT DNA CONSTRUCTS AND RECOMBINANT MICROORGANISMS CARRYING THIS SEQUENCE 有权DNA序列编码派尼列腺素酶，新型重组DNA构建体和携带此序列的重组微生物公开(公告)号：US20100112673A1
公开(公告)日：2010-05-06
申请号：US12524713
申请日：2007-05-15
Applicant: Pavel Kyslik , Vaclav Stepanek , Lenka Hollerova , Stanislav Becka , Vyasarayani Williams Rajasekar , Datla Anupama , Kamila Plhackova , Jaroslav Marsalek
Inventor： Pavel Kyslik , Vaclav Stepanek , Lenka Hollerova , Stanislav Becka , Vyasarayani Williams Rajasekar , Datla Anupama , Kamila Plhackova , Jaroslav Marsalek
CPC classification number: C12N9/84 , C12Y305/01011
Abstract: The invention consists in a nucleotide sequence having the size of (2646) bp, wherein the order of nucleotides is identical to the order of the nucleotide sequence encoding penicillin acylase from Achromobacter sp. CCM 4824 (formerly Comamonas testosteroni CCM 4824), eventually of the fragments of this sequence having the length of at least 150 nucleotides. The sequence can be used in the formation of a DNA construct, eventually the construct having at least one regulatory sequence regulating the expression of the gene and the production of a polypeptide with the penicillin acylase activity. The sequence can form part of a recombinant expression vector, which consists of the above-mentioned construct, promoter, translational start signal, translational and transcriptional stop signal. Further, the invention concerns a recombinant host cell, containing the nucleic acid construct carried by the vector or integrated into the cell chromosome, and the E. coli BL21 strain containing said sequence of the nucleotides encoding the penicillin acylase carried in the pKXIP1, the pKLP3 or the pKLP6 plasmid.
Abstract translation: 本发明包括具有（2646）bp大小的核苷酸序列，其中核苷酸的顺序与编码来自无色杆菌属的青霉素酰基转移酶的核苷酸序列的顺序相同。 CCM 4824（以前的Comamonas testosteroni CCM 4824），最终具有长度至少为150个核苷酸的该序列的片段。 该序列可以用于DNA构建体的形成，最终具有至少一个调节基因表达调节序列和具有青霉素酰基转移酶活性的多肽的产生的构建体。 该序列可以形成重组表达载体的一部分，其由上述构建体，启动子，翻译启动信号，翻译和转录停止信号组成。 此外，本发明涉及含有载体携带或整合到细胞染色体中的核酸构建体的重组宿主细胞，以及含有编码pKXIP1携带的青霉素酰基转移酶的核苷酸序列的大肠杆菌BL21菌株，pKLP3 或pKLP6质粒。
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2.发明授权DNA sequence encoding penicillin acylase, novel recombinant recombinant DNA constructs and recombinant microorganisms 有权编码青霉素酰基转移酶的DNA序列，新型重组重组DNA构建体和重组微生物公开(公告)号：US08039604B2
公开(公告)日：2011-10-18
申请号：US12524713
申请日：2007-05-15
Applicant: Pavel Kyslik , Vaclav Stepanek , Lenka Hollerova , Stanislav Becka , Vyasarayani Williams Rajasekar , Datla Anupama , Kamila Plhackova , Jaroslav Marsalek
Inventor： Pavel Kyslik , Vaclav Stepanek , Lenka Hollerova , Stanislav Becka , Vyasarayani Williams Rajasekar , Datla Anupama , Kamila Plhackova , Jaroslav Marsalek
CPC classification number: C12N9/84 , C12Y305/01011
Abstract: The invention consists in a nucleotide sequence having the size of (2646) bp, wherein the order of nucleotides is identical to the order of the nucleotide sequence encoding penicillin acylase from Achromobacter sp. CCM 4824 (formerly Comamonas testosteroni CCM 4824), eventually of the fragments of this sequence having the length of at least 150 nucleotides. The sequence can be used in the formation of a DNA construct, eventually the construct having at least one regulatory sequence regulating the expression of the gene and the production of a polypeptide with the penicillin acylase activity. The sequence can form part of a recombinant expression vector, which consists of the above-mentioned construct, promoter, translational start signal, translational and transcriptional stop signal. Further, the invention concerns a recombinant host cell, containing the nucleic acid construct carried by the vector or integrated into the cell chromosome, and the E. coli BL21 strain containing said sequence of the nucleotides encoding the penicillin acylase carried in the pKXIP1, the pKLP3 or the pKLP6 plasmid.
Abstract translation: 本发明包括具有（2646）bp大小的核苷酸序列，其中核苷酸的顺序与编码来自无色杆菌属的青霉素酰基转移酶的核苷酸序列的顺序相同。 CCM 4824（以前的Comamonas testosteroni CCM 4824），最终具有长度至少为150个核苷酸的该序列的片段。 该序列可以用于DNA构建体的形成，最终具有至少一个调节基因表达调节序列和具有青霉素酰基转移酶活性的多肽的产生的构建体。 该序列可以形成重组表达载体的一部分，其由上述构建体，启动子，翻译启动信号，翻译和转录停止信号组成。 此外，本发明涉及含有载体携带或整合到细胞染色体中的核酸构建体的重组宿主细胞，以及含有编码pKXIP1携带的青霉素酰基转移酶的核苷酸序列的大肠杆菌BL21菌株，pKLP3 或pKLP6质粒。
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3.发明申请PROCESS FOR THE PREPARATION OF IMMOBILIZED RECOMBINANT PENICILLIN ACYLASE CATALYST FROM ACHROMOBACTER SP. CCM 4824 EXPRESSED IN E. COLI BL 21 CCM 7394 AND ITS USE FOR THE SYNTHESIS OF BETA-LACTAM ANTIBIOTICS 审中-公开用于制备来自ACHROMOBACTER SP的固定化重组苯丙氨酸乙酰化酶催化剂的方法 CCM 4824在大肠杆菌BL21中表达CCM 7394及其用于合成BETA-LACTAM抗生素的用途公开(公告)号：US20100190206A1
公开(公告)日：2010-07-29
申请号：US12670792
申请日：2007-07-27
Applicant: Anupama Datla , Vyasarayani Williams Rajasekar , Pavel Kyslik , Stanislav Becka , Ashar Trupti Krishnakant , Zambre Sujata Yogesh , Kumar Nikunj
Inventor： Anupama Datla , Vyasarayani Williams Rajasekar , Pavel Kyslik , Stanislav Becka , Ashar Trupti Krishnakant , Zambre Sujata Yogesh , Kumar Nikunj
Abstract: The present invention discloses isolation of Penicillin Acylase (PA) from Achromobacter sp CCM 4824 expressed in recombinant strain E. coli BL21 CCM 7394 bearing the recombinant plasmid pKXIP1 and processing of PA into biocatalyst useful for the industrial synthesis of antibiotics. More particularly the invention discloses a synthesis of semi-synthetic β-lactam antibiotics in the reaction mixture consisting of activated acyl-donor (D-p-hydroxyphenylglycine methyl ester or amide for Amoxicillin and Cefadroxil; D-phenylglycine methyl ester or amide for Ampicillin and Cephalexin) and nucleophile (6-APA or 7-ADCA) catalyzed by PA obtained from recombinant E. coli BL21 CCM 7394 as the biocatalyst.
Abstract translation: 本发明公开了在具有重组质粒pKXIP1的重组菌大肠杆菌BL21 CCM7394中表达的来自无色杆菌sp CCM 4824的青霉素酰基酶（PA）的分离，并将PA加工成用于工业合成抗生素的生物催化剂。 更具体地，本发明公开了在由活化的酰基供体（Dp-羟基苯基甘氨酸甲酯或阿莫西林和头孢羟氨苄的酰胺）组成的反应混合物中合成半合成的β-内酰胺抗生素; D-苯基甘氨酸甲酯或氨苯青霉素和头孢氨苄的酰胺 ）和由重组大肠杆菌BL21 CCM 7394作为生物催化剂获得的PA催化的亲核试剂（6-APA或7-ADCA）。
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