MICROBE THAT PRODUCES CAROTENOID
    21.
    发明公开

    公开(公告)号:EP4328317A1

    公开(公告)日:2024-02-28

    申请号:EP22787940.0

    申请日:2022-03-18

    申请人: ENEOS Corporation

    发明人: KUWAHARA Daichi

    摘要: The object of the present invention is to provide a production method for β-cryptoxanthin with improved production efficiency. The present invention provides a method for producing β-cryptoxanthin by culturing bacteria of the genus Paracoccus at a given dissolved oxygen concentration, wherein the bacteria of the genus Paracoccus are bacteria forming yellow to orange colonies; or bacteria whose endogenous crtW gene has been partially or completely deleted or inactivated to lose the ability to produce a functional crtW enzyme.

    SYNTHESIS OF FUNCTIONAL CYTOCHROME P450 MONOOXYGENASES USING A EUKARYOTIC CELL-FREE SYSTEM COMPRISING MEMBRANE VESICLES

    公开(公告)号:EP4328302A1

    公开(公告)日:2024-02-28

    申请号:EP22192384.0

    申请日:2022-08-26

    IPC分类号: C12N9/02 C12P21/02

    摘要: The invention relates to a method for preparing functional membrane attached monooxygenases in cell-free systems which comprises at least the following steps:
    a) providing a reaction medium comprising at least the following components:
    a nucleic acid template coding for a Cytochrome P450 monooxygenase classified as EC 1.14.-.-, an RNApolymerase, a eukaryotic cell lysate containing ER-derived membrane vesicles which comprise NADPH-Cytochrom-P450-oxidoreductase (CPR), optionally heme or heme derivatives, in particular heme complexes selected from the group comprising or consisting of hematin and hemin;
    b) performing a cell-free reaction to synthesize the monooxygenase protein, wherein the cell-free translation reaction is performed at a temperature in the range from 18 to 33°C.
    The invention further relates to a reaction medium for preparing a functional Cytochrome P450 monooxygenase classified as EC 1.14.-.- in a cell-free system comprising at least the following components:
    a nucleic acid template coding for a Cytochrome P450 classified as EC 1.14.-.-, an RNA polymerase, a eukaryotic cell lysate containing membrane vesicles which comprise NADPH-Cytochrom-P450-oxidoreductase (CPR), heme or heme derivatives, in particular heme complexes selected from the group comprising or consisting of hematin and hemin.