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

Inventor： 한예선 ,  고훈영 ,  임혜원 ,  차주환 ,  김욱현 ,  백정호

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。

公开(公告)号：KR1020040011882A

公开(公告)日：2004-02-11

申请号：KR1020020045165

申请日：2002-07-31

Applicant: 한국과학기술연구원

Inventor： 한예선 ,  한수진 ,  백정호 ,  신평균 ,  정찬성 ,  성문희 ,  조현성 ,  이병석

IPC: C12N15/55

CPC classification number: C12N9/20 ,  C07K14/212 ,  C12N15/75 ,  C12Y301/01003

Abstract: PURPOSE: Provided are a gene coding lipase of Acinetobacter species SY-01 with stereoselective substrate specificity and a protein expressed therefrom. CONSTITUTION: A gene is characterized by coding amino acid sequence of lipase of Acinetobacter species SY-01, represented by the SEQ ID NO:2. A transformed Bacillus subtilis 168/pSYLipSM(1) (KCTC 10267BP) is obtained by transformation with a recombinant vector plasmid pSYLipSM(1) containing the gene coding the lipase of Acinetobacter species SY-01. The lipase of Acinetobacter species SY-01 is manufactured by the steps of culturing the transformed bacillus subtilis, and isolating and purifying expressed lipase of Acinetobacter species SY-01 from the cultured cell.

Abstract translation: 目的：提供具有立体选择底物特异性的不动杆菌属SY-01的脂肪酶基因和由其表达的蛋白质。 构成：基因的特征在于编码由SEQ ID NO：2表示的不动杆菌属物种SY-01的脂肪酶的氨基酸序列。 通过用含有编码不动杆菌属SY-01的脂肪酶的基因的重组载体质粒pSYLipSM（1）转化获得转化的枯草芽孢杆菌168 / pSYLipSM（1）（KCTC 10267BP）。 通过培养转化的枯草芽孢杆菌并从培养的细胞中分离和纯化不动杆菌属SY-01的表达脂肪酶的步骤制造不动杆菌属物种SY-01的脂肪酶。

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。

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

Inventor： 한예선 ,  고훈영 ,  임혜원 ,  차주환 ,  김욱현 ,  백정호

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。

公开(公告)号：KR1020010046020A

公开(公告)日：2001-06-05

申请号：KR1019990049591

申请日：1999-11-10

Applicant: 한국과학기술연구원

Inventor： 한예선

IPC: C12N15/52 ,  C12N15/70

CPC classification number: C12N9/93 ,  C12Q1/6876 ,  C12Q2521/501

Abstract: PURPOSE: Provided is a DNA ligase-coding gene of hyperthermophilic Aquifex pyrophilus. And protein expressed therefrom is also provided, which is stable at high temperatures and useful for assay methods in medical industry. CONSTITUTION: The DNA ligase-coding gene of hyperthermophilic Aquifex pyrophilus is obtained by the next steps of: i) cultivating Aquifex pyrophilus to obtain DNA; ii) cutting DNA with restriction enzyme HindIII to obtain small segments; iii) inserting the segments into vector pBluescript KS(±) and obtaining clones having segments of DNA ligase through polymerase chain reaction; iv) separating DNA segments included in the clones, 400bp, on agarose gel and making genome library; v) selecting clones showing positive signal when put on ECL direct system; vi) obtaining DNA from the clones and cutting DNA with restriction enzyme HindIII; vii) performing Southern blotting with DNA segments; viii) obtaining segments showing positive signal and linking the segments with pBluescript KS(±); and ix) determining the base sequence of the clones obtained.

Abstract translation: 目的：提供超嗜热Aquifex pyrophilus的DNA连接酶编码基因。 并且还提供了由其表达的蛋白质，其在高温下是稳定的并且可用于医疗行业中的测定方法。 构成：通过下列步骤获得超嗜热水Aqu螨的DNA连接酶编码基因：i）培养Aquifex pyrophilus以获得DNA; ii）用限制酶HindIII切割DNA以获得小片段; iii）将片段插入载体pBluescript KS（±）中，通过聚合酶链反应获得具有DNA连接酶片段的克隆; iv）将克隆中包含的DNA片段（400bp）分离在琼脂糖凝胶上并制备基因组文库; v）选择放在ECL直接系统上时显示阳性信号的克隆; vi）从克隆获得DNA并用限制酶HindIII切割DNA; vii）用DNA片段进行Southern印迹; viii）获得显示阳性信号的片段并将片段与pBluescript KS（±）连接; 和ix）确定获得的克隆的碱基序列。

公开(公告)号：KR100183362B1

公开(公告)日：1999-04-01

申请号：KR1019970001140

申请日：1997-01-16

Applicant: 한국과학기술연구원

Inventor： 한예선 ,  유연규 ,  김성호 ,  임재환 ,  류재련 ,  최인걸

IPC: C12N15/53

CPC classification number: C12N9/0089 ,  A61K38/00

Abstract: 초고온 미생물인 아퀴펙스 파이로필러스(Aquifex pyrophilus)의 슈퍼옥사이드 디스뮤타제 유전자 서열 및 그 서열로부터 발현되는 단백질이 제공된다.
슈퍼옥사이드 디스뮤타제는 산소에 의한 세포의 손상을 막아주는 중요한 방어 기작에 관여하므로, 염증, 자기 면역 질환 및 염색체 손상등의 치료에 의약으로 사용될 수 있다. 특히, 에이. 파이로필러스는 최적 온도 85℃에서 성장하는 초내열성 세균의 일종이므로, 그의 슈퍼옥사이드 디스뮤타제의 열안정성도 다른 미생물에 비해 아주 높아 제약 분야에서 폭 넓게 응용될 수 있다.