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1.发明公开BAUELEMENT AUF BASIS EINES STRUKTURIERBAREN SUBSTRATS MIT DREIDIMENSIONALER, POREN IM NM-BEREICH AUFWEISENDER MEMBRANSTRUKTUR UND HALBLEITERTECHNOLOGISCHES VERFAHREN ZU DESSEN HERSTELLUNG 审中-公开BAUELEMENT AUF BASIS EINES STRUKTURIERBAREN SUBSTRATS MIT DREIDIMENSIONALER，POREN IM NM-BEREICH AUFWEISENDER MEMBRANSTRUKTUR UND HALBLEITERTECHNOLOGISCHES VERFAHREN ZU DESSEN HERSTELLUNG
公开(公告)号：EP3213068A1
公开(公告)日：2017-09-06
申请号：EP15784646.0
申请日：2015-10-21
发明人： LISEC, Thomas , NEBLING, Eric , KUBICK, Stefan , RUPP, Steffen
IPC分类号： G01N33/487 , B81C1/00 , B01D67/00
CPC分类号： G01N33/48721 , B01D67/0039 , B81B2201/0214 , B81B2201/051 , B81B2203/0127 , B81B2203/0353 , B81C1/00087 , G01N27/44791
摘要： The invention relates to a component comprising a carrier composed of a structurable material with at least one through-opening which is closed off by a porous membrane, characterized in that the porous membrane projects from the surface of the component surrounding the through-opening. In many embodiments, the component also comprises a carrier substrate, wherein one side of the carrier substrate which points towards the component and the opposite side of the component preferably form a fluid duct, wherein in the preferred case the at least one through-opening of the carrier communicates on its non-closed side with the fluid duct. The component according to the invention is suitable for installation and for electrochemical measurement of trans-membrane proteins, preferably in double-lipid layers. The invention also proposes various methods for manufacturing the component.
摘要翻译： 本发明涉及一种包括由可结构化材料构成的载体的部件，该结构化材料具有至少一个通过多孔膜封闭的通孔，其特征在于，多孔膜从围绕通孔的部件表面突出。 在许多实施例中，部件还包括载体基板，其中载体基板的指向部件的一侧和部件的相对侧优选地形成流体管道，其中在优选情况下，至少一个通孔 载体在其非封闭侧与流体导管连通。 根据本发明的组件适合于安装和用于跨膜蛋白质的电化学测量，优选在双脂质层中。 本发明还提出了用于制造该部件的各种方法。
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2.发明公开
公开(公告)号：EP4381054A1
公开(公告)日：2024-06-12
申请号：EP22761079.7
申请日：2022-07-29
CPC分类号： C12Y601/01 , C12N9/93 , C12P21/02 , C12Y601/01001 , C12Y601/01004 , C12Y601/01026 , C12N9/127 , C12N9/12
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4.发明公开
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
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.-
5.发明公开
公开(公告)号：EP4245856A1
公开(公告)日：2023-09-20
申请号：EP22162240.0
申请日：2022-03-15
发明人： WALTER, Ruben , KUBICK, Stefan , ZEMELLA, Anne
摘要： The invention relates to a method for preparing a soluble and functional peroxygenase 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 fungal unspecific peroxygenase (UPO) classified as EC 111.2.1, an RNApolymerase, a eukaryotic cell lysate derived from insect cells or mammalian cells and supplemented with glutathione and glutathione-disulfide during lysate preparation wherein the cell lysate further comprises non-fungal membrane vesicles, as well as heme or a heme derivative, in particular a heme complex selected from the group comprising or consisting of hematin and hemin;
b) performing a cell-free reaction to synthesize the peroxygenase protein.
The invention further relates to a method for screening and characterization of mutants or variants of unspecific peroxygenases classified as EC 1.11.2.1., which method comprises steps a)- and b) of the method above and additionally at least a step c) screening the functional peroxygenase obtained after step b) for a target property, such as substrate specificity or stereoselectivity.
A still further aspect of the invention relates to a reaction medium for preparing a soluble and functional peroxygenase in acell-free system comprising at least the following components: a nucleic acid template coding for a fungal unspecific peroxygenase (UPO) classified as EC 111.2.1, an RNApolymerase, a eukaryotic cell lysate derived from insect cells or mammalian cells and supplemented with glutathione and glutathione-disulfide during lysate preparation wherein the cell lysate further comprises non-fungal membrane vesicles and heme or a heme derivative, in particular a heme complex selected from the group comprising or consisting of hematin and hemin.-
6.发明公开VERFAHREN UND VORRICHTUNG ZUR ZELLFREIEN PROTEINSYNTHESE IN GEGENWART EINES CASPASE-INHIBITORS 审中-公开方法和设备，在一个半胱氨酸蛋白酶抑制剂的存在无细胞蛋白质合成
公开(公告)号：EP3049530A2
公开(公告)日：2016-08-03
申请号：EP14777515.9
申请日：2014-09-17
CPC分类号： C12P21/00 , C07K5/0808 , C07K5/1016 , C07K5/1021 , C12M23/34 , C12M29/04 , C12P21/02
摘要： The invention relates to an improved cell-free protein synthesis method using a eukaryotic cell lysate in the presence of a caspase inhibitor, to a device for carrying out said method, and to the use of a caspase inhibitor for increasing the protein yield or for increasing the stability of the synthesized proteins in such a cell-free protein synthesis method using a eukaryotic cell lysate. In a preferred embodiment of the invention, the cell-free protein synthesis method is carried out as a continuous method in a device which comprises at least two compartments separated by a dialysis membrane. The translation reaction takes place in at least one first compartment, the reaction compartment, and during the translation reaction i) reactants are diffused from at least one other compartment, the supply and discharge compartment, into the reaction compartment, and ii) reaction byproducts are diffused from the reaction compartment into the supply and discharge compartment by means of the dialysis membrane. According to the invention, the caspase inhibitor is provided at least in the reaction compartment but can also be preferably provided in the supply and discharge compartment in order to supply unused inhibitors to the reaction compartment.
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