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1.发明授权
公开(公告)号：KR100522133B1
公开(公告)日：2005-10-18
申请号：KR1020020004657
申请日：2002-01-26
Applicant: 한국과학기술연구원
IPC: C12N9/02
CPC classification number: C12N9/0006
Abstract: 본 발명은 효모의 한 종류인 클루이베로마이세스 마르시아누스
(
Kluyveromyces marxianus )로부터 분리, 정제하여 얻은 카르보닐 환원효소 (Carbonyl Reaductase, CR) 및 이를 분리, 정제하는 방법에 관한 것이다. 본 발명에 의하여 얻어진 환원효소에 의한 환원반응은 미생물에 의한 환원반응 및 화학적 합성방법으로 얻어진 환원효소에 의한 환원반응보다 입체선택성이 높고, 비용과 시간이적인 측면에서 우수하다.-
2.发明授权
公开(公告)号：KR100419592B1
公开(公告)日：2004-02-19
申请号：KR1020010074837
申请日：2001-11-29
Applicant: 한국과학기술연구원
IPC: C12N15/53
CPC classification number: C12N9/0036
Abstract: The invention is a gene coding for a quinone oxidoreductase from Kluyveromyces marxianus and a protein having an amino acid sequence expressed therefrom, which can be advantageously used in a reduction reaction of a quinone compound and synthesis of intermediates for a biologically active compound.
Abstract translation: 本发明是编码来自马克斯克鲁维酵母的醌氧化还原酶的基因和具有由其表达的氨基酸序列的蛋白质，其可以有利地用于醌化合物的还原反应和用于生物活性化合物的中间体的合成。
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公开(公告)号：KR1020030044179A
公开(公告)日：2003-06-09
申请号：KR1020010074837
申请日：2001-11-29
Applicant: 한국과학기술연구원
IPC: C12N15/53
CPC classification number: C12N9/0036
Abstract: PURPOSE: A quinone oxidoreductase gene from Kluyveromyces marxianus is provided to be used for the reduction reaction of the quinone compounds and for the intermediate synthesis of the biologically active compounds by way of the excellent quinone reduction activity thereof. CONSTITUTION: A gene codes the amino acid sequence of Kluyveromyces marxianus quinone oxidoreductase(kmQOR) presented at the sequence No. 2. The kmQOR has a molecular weight of about 42kDa. A recombinant vector contains the gene coded with the kmQOR, and is determined to be plasmid pQOR22b. An E. coli is transformed into the recombinant vector. In a method of preparing the kmQOR, the transformed E. coli is first cultured, and β-D-isopropyl-D-thiogalactopyranoside (IPTG) is then added to the cultured E. coli to induce the expression of kmQOR therefrom. The expressed kmQOR is collected, and purified.
Abstract translation: 目的：提供马克斯克鲁维酵母的醌氧化还原酶基因，用于醌类化合物的还原反应和生物活性化合物的中间合成，其优异的醌还原活性。 构成：A基因编码序列号2中提供的马克斯基黄酮醌氧化还原酶（kmQOR）的氨基酸序列。kmQOR的分子量约为42kDa。 重组载体含有用kmQOR编码的基因，并确定为质粒pQOR22b。 将大肠杆菌转化入重组载体。 在制备kmQOR的方法中，首先培养转化的大肠杆菌，然后将β-D-异丙基-D-硫代吡喃半乳糖苷（IPTG）加入到培养的大肠杆菌中以诱导其表达KmQOR。 收集并纯化表达的kmQOR。
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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
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。
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