公开(公告)号：KR100479902B1

公开(公告)日：2005-03-31

申请号：KR1020030018093

申请日：2003-03-24

Applicant: 한국과학기술연구원

Inventor： 한예선 ,  한수진 ,  정찬성 ,  임혜원 ,  조현성 ,  이병석 ,  양은경

IPC: C12N15/55

Abstract: 본 발명은 아시네토박토 종 SY-01의 입체선택적 기질 특이성을 갖는 리파제 변이체의 유전자 서열 및 이로부터 발현되는 단백질에 관한 것이다. 본 발명의 아시네토박토 종 SY-01 리파제로부터 돌연변이된 리파제는 활성이 우수하므로 상기 리파제 변이체 코딩 유전자를 포함하는 형질전환된 리파제를 생체 촉매로서 산업적으로 이용될 수 있다.

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：KR100468547B1

公开(公告)日：2005-01-29

申请号：KR1020020045165

申请日：2002-07-31

Applicant: 한국과학기술연구원

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

IPC: C12N15/55

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：提供具有立体选择性底物特异性的不动杆菌种SY-01的编码脂肪酶的基因和由其表达的蛋白质。 组成：基因的特征在于编码不动杆菌属物种SY-01的脂肪酶的氨基酸序列，由SEQ ID NO：2表示。 通过用含有编码不动杆菌属物种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）。 不动杆菌菌种SY-01的脂肪酶是通过培养转化的枯草芽孢杆菌，从培养细胞中分离纯化不动杆菌菌种SY-01的表达脂肪酶。

公开(公告)号：KR100453212B1

公开(公告)日：2004-10-15

申请号：KR1020020083097

申请日：2002-12-24

Applicant: 한국과학기술연구원

Inventor： 정찬성 ,  이소하 ,  김영희 ,  채한국 ,  조현성

IPC: C12P41/00

Abstract: PURPOSE: A catalyzed resolution method of racemic cis-1,3-dioxolane derivatives is provided, thereby effectively separating racemic cis-1,3-dioxolane derivatives which is useful as an intermediate used for preparing itraconazole and ketoconazole. CONSTITUTION: The catalyzed resolution method of racemic cis-1,3-dioxolane derivatives represented by formula (1) comprises hydrolysis of racemic cis-1,3-dioxolane derivatives using enzyme in a phosphate buffer solution at pH 6 to 9 and 20 to 60 deg. C to prepare pure cis-1,3-dioxolane derivatives of formulas (1a) and (1b), wherein R is CH2OH, COOH, COOR' or CH2OCOR'; R' is C1-C10 alkyl; the enzyme is Pseudomonas cepacia(PCL) or Candida antartica(CAL).

Abstract translation: 目的：提供外消旋顺式-1,3-二氧戊环衍生物的催化拆分方法，由此有效地分离用作制备伊曲康唑和酮康唑中间体的外消旋顺式-1,3-二氧戊环衍生物。 构成：由式（1）表示的外消旋顺式-1,3-二氧戊环衍生物的催化拆分方法包括使用酶在pH 6至9和20至60的磷酸盐缓冲溶液中水解外消旋顺式-1,3-二氧戊环衍生物 度。 制备式（1a）和（1b）的纯顺式-1,3-二氧戊环衍生物，其中R为CH 2 OH，COOH，COOR'或CH 2 OCOR'; R'是C 1 -C 10烷基; 该酶是洋葱假单胞菌（Pseudomonas cepacia）（PCL）或南极假丝酵母（Candida antartica）（CAL）。

公开(公告)号：KR1020040083588A

公开(公告)日：2004-10-06

申请号：KR1020030018093

申请日：2003-03-24

Applicant: 한국과학기술연구원

Inventor： 한예선 ,  한수진 ,  정찬성 ,  임혜원 ,  조현성 ,  이병석 ,  양은경

IPC: C12N15/55

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

Abstract: PURPOSE: Provided are a gene encoding mutant lipase of acinetobacter species SY-01, Bacillus subtilis 168/pSYLipM58 (KCTC 10422BP) as a transformant containing the same, and mutant lipase expressed therefrom displaying improved enzyme activity. The transformant is useful as a bio-catalyst. CONSTITUTION: The gene encoding mutant lipase of Acinetobacter species SY-01 has the nucleotide sequence set forth in SEQ ID NO:2. A recombinant vector of Acinetobacter species SY-01, plasmid pSYLipM58, comprises the gene encoding mutant lipase of Acinetobacter species SY-01. The transformant, Bacillus subtilis 168/pSYLipM58(KCTC 10422BP), contains the plasmid pSYLipM58. The method for producing the mutant lipase of Acinetobacter species SY-01 comprises the steps of: culturing Bacillus subtilis 168/pSYLipM58(KCTC 10422BP); and recovering and purifying the mutant lipase of Acinetobacter species SY-01 expressed from the cultured microorganism.

Abstract translation: 目的：提供编码不动杆菌属SY-01，枯草芽孢杆菌168 / pSYLipM58（KCTC10422BP）的突变体脂肪酸的基因，其为含有该不动杆菌属物种的转化体，由表达改良的酶活性的突变体脂肪酶表达。 转化体可用作生物催化剂。 构成：编码不动杆菌属SY-01的突变体脂肪酶的基因具有SEQ ID NO：2所示的核苷酸序列。 不动杆菌属SY-01，质粒pSYLipM58的重组载体包含编码不动杆菌属SY-01突变体脂肪酶的基因。 转化体枯草芽孢杆菌168 / pSYLipM58（KCTC10422BP）含有质粒pSYLipM58。 生产不动杆菌属SY-01突变型脂肪酶的方法包括以下步骤：培养枯草芽孢杆菌168 / pSYLipM58（KCTC 10422BP）; 并回收和纯化由培养的微生物表达的不动杆菌属SY-01的突变脂肪酶。

公开(公告)号：KR1020040006307A

公开(公告)日：2004-01-24

申请号：KR1020020040325

申请日：2002-07-11

Applicant: 한국과학기술연구원 ,  한올바이오파마주식회사 ,  경동제약 주식회사

Inventor： 정찬성 ,  한예선 ,  윤문영 ,  조현성 ,  이병석

IPC: C12P17/04

Abstract: PURPOSE: A process for preparing cis-2- (bromomethyl)-2- (2,4-dichlorophenyl)- 13-dioxolane- 4-methyl alcohol having high optical selectivity is provided. A stereo-selective enzyme maintains its maximum activity because the reaction product is rapidly removed from the reaction solution during the reaction in order not to inhibit the activity of the enzyme. CONSTITUTION: A process for preparing cis-2- (bromomethyl)-2- (2,4-dichlorophenyl)- 13-dioxolane- 4-methyl alcohol having high optical selectivity comprises hydrolysis of cis-2- (bromomethyl)-2- (2,4-dichlorophenyl)- 1,3-dioxolane- 4-methyl acetate using lipase produced from Acinetobacter sp. SY-01(KFCC-11111), and recovering an alcohol product as a reaction product from the reaction solution during the reaction, wherein the concentration of the alcohol product is 0.1 to 300 mg/l; the reaction product is recovered when the conversion rate of the hydrolysis reaction is 40 to 60%; the hydrolysis reaction is carried out under conditions of 40 to 60 deg. C, pH 7.0 to 9.0 and 100 to 300 rpm; and the reaction product is recovered by column chromatography.

Abstract translation: 目的：提供具有高光学选择性的顺式-2-（溴甲基）-2-（2,4-二氯苯基）-1,3-二氧戊环-4-甲醇的方法。 立体选择性酶维持其最大活性，因为在反应期间快速从反应溶液中除去反应产物，以便不抑制酶的活性。 构成：具有高光学选择性的顺式-2-（溴甲基）-2-（2,4-二氯苯基）-1,3-二氧戊环-4-甲醇的制备方法包括顺式-2-（溴甲基）-2-（ 2,4-二氯苯基）-1,3-二氧戊环-4-甲基乙酸酯，使用由不动杆菌属（Acinetobacter sp。）生产的脂肪酶。 SY-01（KFCC-11111），并且在反应期间从反应溶液中回收作为反应产物的醇产物，其中醇产物的浓度为0.1至300mg / l; 当水解反应的转化率为40〜60％时回收反应产物; 水解反应在40〜60℃的条件下进行。 C，pH 7.0至9.0和100至300rpm; 并通过柱色谱法回收反应产物。

公开(公告)号：KR100446256B1

公开(公告)日：2004-09-01

申请号：KR1020020040325

申请日：2002-07-11

Applicant: 한국과학기술연구원 ,  한올바이오파마주식회사 ,  경동제약 주식회사

Inventor： 정찬성 ,  한예선 ,  윤문영 ,  조현성 ,  이병석

IPC: C12P17/04

Abstract: PURPOSE: A process for preparing cis-2- (bromomethyl)-2- (2,4-dichlorophenyl)- 13-dioxolane- 4-methyl alcohol having high optical selectivity is provided. A stereo-selective enzyme maintains its maximum activity because the reaction product is rapidly removed from the reaction solution during the reaction in order not to inhibit the activity of the enzyme. CONSTITUTION: A process for preparing cis-2- (bromomethyl)-2- (2,4-dichlorophenyl)- 13-dioxolane- 4-methyl alcohol having high optical selectivity comprises hydrolysis of cis-2- (bromomethyl)-2- (2,4-dichlorophenyl)- 1,3-dioxolane- 4-methyl acetate using lipase produced from Acinetobacter sp. SY-01(KFCC-11111), and recovering an alcohol product as a reaction product from the reaction solution during the reaction, wherein the concentration of the alcohol product is 0.1 to 300 mg/l; the reaction product is recovered when the conversion rate of the hydrolysis reaction is 40 to 60%; the hydrolysis reaction is carried out under conditions of 40 to 60 deg. C, pH 7.0 to 9.0 and 100 to 300 rpm; and the reaction product is recovered by column chromatography.

Abstract translation: 目的：提供一种制备光学选择性高的顺式-2-（溴甲基）-2-（2,4-二氯苯基）-13-二氧戊环-4-甲醇的方法。 立体选择性酶保持其最大活性，因为在反应过程中反应产物从反应溶液中快速除去以不抑制酶的活性。 结构：制备具有高光学选择性的顺式-2-（溴甲基）-2-（2,4-二氯苯基）-13-二氧戊环-4-甲醇的方法包括顺式-2-（溴甲基）-2-（ 2,4-二氯苯基）-1,3-二氧戊环-4-甲基乙酸酯使用由不动杆菌产生的脂肪酶进行。 SY-01（KFCC-11111），在反应过程中从反应溶液中回收作为反应产物的醇产物，其中醇产物的浓度为0.1-300mg / l; 当水解反应的转化率为40至60％时，反应产物被回收; 水解反应在40〜60℃的条件下进行。 C，pH 7.0至9.0和100至300rpm; 并通过柱色谱法回收反应产物。

公开(公告)号：KR1020040056592A

公开(公告)日：2004-07-01

申请号：KR1020020083097

申请日：2002-12-24

Applicant: 한국과학기술연구원

Inventor： 정찬성 ,  이소하 ,  김영희 ,  채한국 ,  조현성

IPC: C12P41/00

Abstract: PURPOSE: A catalyzed resolution method of racemic cis-1,3-dioxolane derivatives is provided, thereby effectively separating racemic cis-1,3-dioxolane derivatives which is useful as an intermediate used for preparing itraconazole and ketoconazole. CONSTITUTION: The catalyzed resolution method of racemic cis-1,3-dioxolane derivatives represented by formula (1) comprises hydrolysis of racemic cis-1,3-dioxolane derivatives using enzyme in a phosphate buffer solution at pH 6 to 9 and 20 to 60 deg. C to prepare pure cis-1,3-dioxolane derivatives of formulas (1a) and (1b), wherein R is CH2OH, COOH, COOR' or CH2OCOR'; R' is C1-C10 alkyl; the enzyme is Pseudomonas cepacia(PCL) or Candida antartica(CAL).

Abstract translation: 目的：提供外消旋顺式-1,3-二氧戊环衍生物的催化拆分方法，有效分离外消旋顺式-1,3-二氧戊环衍生物，其用作制备伊曲康唑和酮康唑的中间体。 构成：由式（1）表示的外消旋顺式-1,3-二氧戊环衍生物的催化分解方法包括使用酶在pH6至9和20至60的磷酸盐缓冲溶液中水解外消旋的顺式-1,3-二氧戊环衍生物 度。 制备式（1a）和（1b）的纯的顺式-1,3-二氧戊环衍生物，其中R是CH 2 OH，COOH，COOR'或CH 2 OCOR'; R'是C 1 -C 10烷基; 该酶是洋葱假单胞菌（PCL）或假丝酵母（CAL）。