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    클루이베로마이세스 마르시아누스로부터 분리 및 정제된 카르보닐 환원효소 및 그 제조 방법
    2.
    发明公开
    클루이베로마이세스 마르시아누스로부터 분리 및 정제된 카르보닐 환원효소 및 그 제조 방법 失效
    氧化还原血红蛋白血症和分离及其纯化方法

    公开(公告)号：KR1020030064159A

    公开(公告)日：2003-07-31

    申请号：KR1020020004657

    申请日：2002-01-26

    Applicant: 한국과학기술연구원

    Inventor： 고훈영 ,  한예선 ,  차주환 ,  김욱현 ,  이종수 ,  윤홍철

    IPC: C12N9/02

    CPC classification number: C12N9/0006

    Abstract: PURPOSE: A carbonyl reductase of Kluyveromyces marxianus and an isolation and purification method thereof are provided, thereby higher stereo-selectively, cheaply and rapidly carrying out the reduction process. CONSTITUTION: A carbonyl reductase isolated and purified from Kluyveromyces marxianus has the properties of the size of 40 to 42 kDa, the active temperature of 25 to 35 deg. C, and the active pH of 6.5 to 7.5, wherein the carbonyl reductase contains the amino acid sequence of Thr-Phe-Thr-Val-Val-Thr-Gly in the amino-terminal. A method for isolating and purifying the carbonyl reductase of Kluyveromyces marxianus comprises the steps of: (1) culturing Kluyveromyces marxianus; (2) centrifuging the cultured medium to collect the pellet and pulverizing the pellet; and (3) centrifuging the pulverized cell extract to collect the supernatant, and subjecting the supernatant to column chromatography to isolate an active fraction containing active material to substrate, wherein the column uses Q sepharose, phenyl sepharose, high-trap blue and gel filtration chromatography, sequentially.

    Abstract translation: 目的：提供马克斯克鲁维酵母的羰基还原酶及其分离和纯化方法，从而立体选择性更好，成本低廉且快速地进行还原过程。 构成：从马克斯克鲁维酵母分离和纯化的羰基还原酶具有40至42kDa大小的性质，活性温度为25至35度。 C，活性pH为6.5〜7.5，其中羰基还原酶含有氨基末端的Thr-Phe-Thr-Val-Val-Thr-Gly的氨基酸序列。 分离和纯化马克斯克鲁维酵母的羰基还原酶的方法包括以下步骤：（1）培养马克斯克鲁维酵母; （2）离心培养基以收集颗粒并粉碎颗粒; （3）将粉碎的细胞提取物离心以收集上清液，并将上清液进行柱色谱以将含有活性物质的活性级分与底物分离，其中该柱使用Q琼脂糖凝胶，苯基琼脂糖凝胶，高陷阱蓝和凝胶过滤色谱 ，顺序。

    클루이베로마이시스 마르시아누스의 카르보닐 환원효소를이용한 카르보닐 화합물의 환원반응
    3.
    发明公开
    클루이베로마이시스 마르시아누스의 카르보닐 환원효소를이용한 카르보닐 화합물의 환원반응 失效
    碳化合物和使用从KLUYVEROMYCES MARXIANUS分离的碳氧化还原酶的制备方法

    公开(公告)号：KR1020030056992A

    公开(公告)日：2003-07-04

    申请号：KR1020010087362

    申请日：2001-12-28

    Applicant: 한국과학기술연구원

    Inventor： 고훈영 ,  최경일 ,  조용서 ,  배애님 ,  차주환 ,  한예선 ,  이종수 ,  윤홍철

    IPC: C07D209/48

    CPC classification number: C07D209/48

    Abstract: PURPOSE: Carbonyl compounds and a preparation process thereof using carbonyl reductase isolated from Kluyveromyces marxianus are provided. The carbonyl compounds are useful as intermediates for preparing beta-lactam family antimicrobial agents. CONSTITUTION: Carbonyl compounds represented by the formula(Ia) and (Ib) are provided, wherein R is methyl, ethyl, propyl, isopropyl, isobutyl and allyl containing saturated or unsaturated alkyl, or phenyl containing aryl; R1, R2, R3 and R4 are independently selected from the group consisting of hydrogen, halogen atoms including Br, Cl, F and I, methyl and ethyl containing C1-C4 alkyl, hydroxy, C1-C4 alkoxy, acetoxy containing ester, and phenyl. A process for preparing the carbonyl compound of the formula(Ia) and (Ib) comprises the steps of: mixing a compound of formula(VI) with β-NADPH and a pH 5.0 to 8.0 buffer solution; adding carbonyl reductase isolated from Kluyveromyces marxianus into the mixture; and reacting the mixture at 20 to 40 deg. C for 5 hours to 5 days.

    Abstract translation: 目的：提供羰基化合物及其使用从马克斯克鲁维酵母分离的羰基还原酶的制备方法。 羰基化合物可用作制备β-内酰胺家族抗微生物剂的中间体。 构成：提供由式（Ia）和（Ib）表示的羰基化合物，其中R是含有饱和或不饱和的烷基或含苯基的芳基的甲基，乙基，丙基，异丙基，异丁基和烯丙基; R 1，R 2，R 3和R 4独立地选自氢，包括Br，Cl，F和I的卤素原子，含有C 1 -C 4烷基，羟基，C 1 -C 4烷氧基，含乙酰氧基的酯和苯基的甲基和乙基 。 制备式（Ia）和（Ib）的羰基化合物的方法包括以下步骤：将式（VI）化合物与β-NADPH和pH 5.0-8.0缓冲溶液混合; 将从马克斯克鲁维酵母分离的羰基还原酶加入到混合物中; 并使混合物在20至40℃下反应。 C为5小时至5天。

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