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    Recombinant bioprocess for the preparation of 7-amino cephalosporanic acid (7-ACA)
    1.
    发明授权
    Recombinant bioprocess for the preparation of 7-amino cephalosporanic acid (7-ACA) 失效
    用于制备7-氨基头孢菌酸（7-ACA）的重组生物过程

    公开(公告)号：US5629171A

    公开(公告)日：1997-05-13

    申请号：US439404

    申请日：1995-05-11

    Applicant: Michael J. Conder ,  John A. Rambosek ,  Phyllis C. McAda ,  Christopher D. Reeves

    Inventor： Michael J. Conder ,  John A. Rambosek ,  Phyllis C. McAda ,  Christopher D. Reeves

    IPC: C12N1/19 ,  C07D501/20 ,  C12N1/15 ,  C12N9/00 ,  C12N9/02 ,  C12N9/10 ,  C12N15/09 ,  C12N15/52 ,  C12N15/54 ,  C12P20060101 ,  C12P35/00 ,  C12P35/06 ,  C12P37/04 ,  C12R1/82 ,  C12N1/16 ,  C12P35/02

    CPC classification number: C12N9/0071 ,  C12N9/00 ,  C12N9/1029 ,  C12P35/00 ,  C12P37/04 ,  Y10S435/935

    Abstract: Important intermediates for preparing cephalosporin antibiotics, 7-amino-cephalosporanic acid (7-ACA) and 7-aminodeacetylcephalosporanic acid (7-ADAC), are prepared by a novel bioprocess in which a transformed Penicillium chrysogenum strain is cultured in the presence of an adipate feedstock to produce adipoyl-6-APA (6-amino penicillanic acid); followed by the in situ expression of the following genes with which the P. chrysogenum has been transformed:1) an expandase gene, whose expression product converts the adipoyl-6-APA by ring expansion to adipoyl-7-ADCA;2) an hydroxylase gene whose expression product converts the 3-methyl side chain of adipoyl-7-ADCA to 3-hydroxymethyl, to give the first product, 7-aminodeacetylcephalosporanic acid (7-ADAC); and3) an acetyltransferase gene whose expression product converts the 3-hydroxymethyl side chain to the 3-acetyloxymethyl side chain of 7-ACA. The final product, 7-ACA, is then prepared by cleavage of the adipoyl side chain using an adipoyl acylase. The entire synthesis, accordingly, is carried out using bioprocesses, and is efficient and economical.

    Abstract translation: 用于制备头孢菌素抗生素，7-氨基 - 头孢菌酸（7-ACA）和7-氨基脱乙酰头孢菌酸（7-ADAC）的重要中间体是通过新型生物工艺制备的，其中转化的产黄青霉菌株在己二酸存在下培养 生产己二酰-6-APA（6-氨基青霉烷酸）的原料; 其次是产生产黄青霉素的以下基因的原位表达：1）扩展酶基因，其表达产物通过环扩展将己二酰基-6-APA转化为己二酰-7-ADCA; 2）表达产物将己二酰-7-ADCA的3-甲基侧链转化为3-羟甲基的羟化酶基因，得到第一产物7-氨基乙酰头孢烷酸（7-ADAC）; 和3）其表达产物将3-羟甲基侧链转化为7-ACA的3-乙酰氧甲基侧链的乙酰转移酶基因。 然后通过使用己二酰酰化酶裂解己二酰侧链制备最终产物7-ACA。 因此，整个合成使用生物工艺进行，并且是高效和经济的。

    Bioprocesses for preparing 7-ACA and 7-ADAC
    4.
    发明授权
    Bioprocesses for preparing 7-ACA and 7-ADAC 失效
    用于制备7-ACA和7-ADAC的生物工艺

    公开(公告)号：US6071713A

    公开(公告)日：2000-06-06

    申请号：US340781

    申请日：1999-06-28

    Applicant: Michael J. Conder ,  John A. Rambosek ,  Phyllis C. McAda ,  Christopher D. Reeves

    Inventor： Michael J. Conder ,  John A. Rambosek ,  Phyllis C. McAda ,  Christopher D. Reeves

    IPC: C12N1/19 ,  C07D501/20 ,  C12N1/15 ,  C12N9/00 ,  C12N9/02 ,  C12N9/10 ,  C12N15/09 ,  C12N15/52 ,  C12N15/54 ,  C12P20060101 ,  C12P35/00 ,  C12P35/06 ,  C12P37/04 ,  C12R1/82 ,  C12N1/14 ,  C12N1/20 ,  C12P35/02

    CPC classification number: C12N9/0071 ,  C12N9/00 ,  C12N9/1029 ,  C12P35/00 ,  C12P37/04 ,  Y10S435/935

    Abstract: A bioprocess for preparing adipoyl-7-ACA comprising the steps: (a) transforming cells of a strain of Penicillium chrysogenum which produces isopenicillin N with an expression vector containing DNA encoding an enzyme, having expandase activity capable of accepting adipoyl 6-APA as a substrate, an enzyme having hydroxylase activity capable of accepting adipoyl-7-ADCA as a substrate and an enzyme having acetyl transferase activity capable of accepting adipoyl 7-ADAC as a substrate; (b) culturing the transformed cells from step a) in a suitable culture medium containing an adipate feedstock, wherein said cells produce adipoyl 6-APA; and (c) culturing the transformed cells producing adipoyl 6-APA of step b) under conditions suitable for expression of said DNA encoding enzyme, thereby producing the end product adipoyl-7-ACA.

    Abstract translation: 一种用于制备己二酰-7-ACA的生物工艺，包括以下步骤：（a）用含有编码酶的DNA的表达载体转化产生异青霉素N的产青霉菌菌株的细胞，其具有能够接受己二酰基6-APA的扩环酶活性 底物，能够接受己二酰-7-ADCA作为底物的羟化酶活性的酶和能够接受己二酰基7-ADAC作为底物的乙酰转移酶活性的酶; （b）在含有己二酸原料的合适培养基中培养来自步骤a）的转化细胞，其中所述细胞产生己二酰基6-APA; 和（c）在适于表达所述DNA编码酶的条件下培养产生步骤b）的己二酰基6-APA的转化细胞，从而产生最终产物己二酰-7-ACA。

    Bioprocess for preparing 7-ACA and 7-ADAC
    9.
    发明授权
    Bioprocess for preparing 7-ACA and 7-ADAC 失效
    用于制备7-ACA和7-ADAC的生物工艺

    公开(公告)号：US5559005A

    公开(公告)日：1996-09-24

    申请号：US250310

    申请日：1994-05-27

    Applicant: Michael J. Conder ,  Phyllis C. McAda ,  John A. Rambosek ,  Christopher D. Reeves

    Inventor： Michael J. Conder ,  Phyllis C. McAda ,  John A. Rambosek ,  Christopher D. Reeves

    IPC: C12N1/19 ,  C07D501/20 ,  C12N1/15 ,  C12N9/00 ,  C12N9/02 ,  C12N9/10 ,  C12N15/09 ,  C12N15/52 ,  C12N15/54 ,  C12P20060101 ,  C12P35/00 ,  C12P35/06 ,  C12P37/04 ,  C12R1/82 ,  C12P35/04 ,  C12N1/16

    CPC classification number: C12N9/0071 ,  C12N9/00 ,  C12N9/1029 ,  C12P35/00 ,  C12P37/04 ,  Y10S435/935

    Abstract: Important intermediates for preparing cephalosporin antibiotics, 7-amino-cephalosporanic acid (7-ACA) and 7-aminodeacetylcephalosporanic acid (7-ADAC), are prepared by a novel bioprocess in which a transformed Penicillium chrysogenum strain is cultured in the presence of an adipate feedstock to produce adipoyl-6-APA (6-amino penicillanic acid); followed by the in situ expression of the following genes with which the P. chrysogenum has been transformed:1) an expandase gene, e.g., from Cephalosporium acremonium, whose expression product converts the adipoyl-6-APA by ring expansion to adipoyl-7-ADCA;2) an hydroxylase gene whose expression product converts the 3-methyl side chain of adipoyl-7-ADCA to 3-hydroxymethyl, to give the first product, 7-aminodeacetylcephalosporanic acid (7-ADAC); and3) an acetyltransferase gene whose expression product converts the 3-hydroxymethyl side chain to the 3-acetyloxymethyl side chain of 7-ACA. The final product, 7-ACA, is then prepared by cleavage of the adipoyl side chain using an adipoyl acylase. The entire synthesis, accordingly, is carried out using bioprocesses, and is efficient and economical.

    Abstract translation: 用于制备头孢菌素抗生素，7-氨基头孢菌素酸（7-ACA）和7-氨基脱乙酰头孢菌酸（7-ADAC）的重要中间体是通过新型生物工艺制备的，其中转化的产黄青霉菌株在己二酸存在下培养 生产己二酰-6-APA（6-氨基青霉烷酸）的原料; 其次是产生产黄青霉的以下基因的原位表达：1）扩增酶基因，例如头孢霉头孢菌，其表达产物通过环扩展将己二酰基-6-APA转化为己二酰-7- ADCA; 2）表达产物将己二酰-7-ADCA的3-甲基侧链转化为3-羟甲基的羟化酶基因，得到第一产物7-氨基乙酰基头孢烷酸（7-ADAC）; 和3）其表达产物将3-羟甲基侧链转化为7-ACA的3-乙酰氧甲基侧链的乙酰转移酶基因。 然后通过使用己二酰酰化酶裂解己二酰侧链制备最终产物7-ACA。 因此，整个合成使用生物工艺进行，并且是高效和经济的。

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