公开(公告)号：KR1020110086686A

公开(公告)日：2011-07-29

申请号：KR1020110071078

申请日：2011-07-18

Applicant: 전남대학교산학협력단

Inventor： 윤철호 ,  김건희 ,  김동현 ,  강지연

IPC: C12N15/09 ,  C12P7/02

Abstract: PURPOSE: A method for preparing human metabolites of simvastatin or lovastatin using bacterial cytochrome p450 is provided to selectively produce metabolic protucts. CONSTITUTION: A composition for preparing human metabolites of simvastatin or lovastatin has: 47th amino acid arginin(R) of wild type CYP102A1, which is substituted with alanine, valine, leucine, isoleucine, proline, methionine, phenylalanine, or tryptopane; 87th amino acid, phenylalaine(F) which is substituted with alanine, valine, leucine, isoleucine, proline, methionine, or tryptopane; 143th amino acid, gluamic acid(E), which is substituted with glycine, serine, threonine, cysteine, tyrosine, asparagine, or glutamine; and 188th amino acid, leucine(L), which is substituted with glycine, serine, threonine, cysteine, tyrosine, asparagine, or glutamine.

Abstract translation: 目的：提供使用细菌细胞色素p450制备辛伐他汀或洛伐他汀的人代谢产物的方法，以选择性地产生代谢产物。 构成：用于制备辛伐他汀或洛伐他汀的人代谢物的组合物具有：用丙氨酸，缬氨酸，亮氨酸，异亮氨酸，脯氨酸，甲硫氨酸，苯丙氨酸或三苯甲烷取代的野生型CYP102A1的第47位氨基酸精氨酸（R） 第八十八氨基酸，被丙氨酸，缬氨酸，亮氨酸，异亮氨酸，脯氨酸，甲硫氨酸或三苯甲烷取代的苯丙氨酸（F）; 由甘氨酸，丝氨酸，苏氨酸，半胱氨酸，酪氨酸，天冬酰胺或谷氨酰胺取代的第143位氨基酸，葡萄糖酸（E） 和第188位氨基酸，其被甘氨酸，丝氨酸，苏氨酸，半胱氨酸，酪氨酸，天冬酰胺或谷氨酰胺取代的亮氨酸（L）。

公开(公告)号：KR101190881B1

公开(公告)日：2012-10-12

申请号：KR1020090078181

申请日：2009-08-24

Applicant: 전남대학교산학협력단

Inventor： 윤철호 ,  김건희 ,  김동현 ,  강지연

IPC: C12N15/09 ,  C12P7/02

CPC classification number: C12P17/06 ,  C12N9/0071 ,  C12Y114/14001

Abstract: 본 발명은 심바스타틴 또는 로바스타틴의 사람에서의 대사산물을 생산하는 방법에 관한 것이다. 보다 상세하게는, 본 발명은 박테리아 시토크롬 P450 BM3(CYP102A1) 또는 그의 돌연변이체를 이용하여 심바스타틴 또는 로바스타틴으로부터 그의 사람에서의 대사산물을 제조하는 방법 및 이를 위한 조성물 및 키트를 제공한다.
심바스타틴, 로바스타틴, CYP102A1, 돌연변이체

公开(公告)号：KR1020090093166A

公开(公告)日：2009-09-02

申请号：KR1020080018534

申请日：2008-02-28

Applicant: 전남대학교산학협력단

Inventor： 윤철호 ,  임성근 ,  김동현 ,  김건희

IPC: C12Q1/32 ,  C12Q1/26 ,  G01N33/52 ,  G01N33/50

CPC classification number: C12Q1/32 ,  G01N2333/90209 ,  C12Y106/02004 ,  G01N33/52

Abstract: A method for measuring the activation of NADPH-cytochrome P450 reductase (CPR) in a sample with 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) is provided to shorten the analysis time and easily and accurately detect a small amount of CTC-formazan. A method for measuring the activation of NADPH-cytochrome P450 reductase (CPR) in a sample comprises: a step of reacting a sample containing an enzyme having CPR activation with 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) under the presence of NADPH to convert the CTC to CTC-formazan; and a step of measuring the concentration of CTC-formazan with spectrophotometric or spectrofluorometric method.

Abstract translation: 提供了一种用5-氰基-2,3-二苯基四唑氯化物（CTC）测定样品中NADPH-细胞色素P450还原酶（CPR）活化的方法，以缩短分析时间，容易且准确地检测少量CTC -formazan。 用于测量样品中NADPH-细胞色素P450还原酶（CPR）的活化的方法包括：将含有CPR活化的酶的样品与5-氰基-2,3-二甲苯基四唑氯化物（CTC）反应的步骤， 的NADPH将CTC转化为CTC-甲an; 并用分光光度法或分光荧光法测定CTC-甲the的浓度。

公开(公告)号：KR1020110020525A

公开(公告)日：2011-03-03

申请号：KR1020090078181

申请日：2009-08-24

Applicant: 전남대학교산학협력단

Inventor： 윤철호 ,  김건희 ,  김동현 ,  강지연

IPC: C12N15/09 ,  C12P7/02

CPC classification number: C12P17/06 ,  C12N9/0071 ,  C12Y114/14001

Abstract: PURPOSE: A method for selective producing human metabolite from simvastatin or lovastatin is provided. CONSTITUTION: A composition for catalysis for producing human metabolite from simvastatin or lovastatin contains wild type CYP102A1 and/or CYP102A1 mutant. The CYP102A1 mutant has 47th amino acid, arginin substituted with leucine; 51th amino acid, tyrosine substituted with phenylalanine; 64th amino acid, glytamic acid substituted with glycine; 74th amino acid, alanine substituted with glycine, 81th amino acid, phenylalanine substituted with isoleucine, 86th amino acid, leucine substituted with isoleucine; 87th amino acid, phenylalanine substituted with valine; 143th amino acid, gluamic acid substituted with glycine; 188th amino acid, leucine substituted with glutamine; and 267th amino acid glutamic acid substituted with valine.

Abstract translation: 目的：提供从辛伐他汀或洛伐他汀选择性生产人类代谢物的方法。 构成：用于从辛伐他汀或洛伐他汀生产人代谢物的催化组合物含有野生型CYP102A1和/或CYP102A1突变体。 CYP102A1突变体具有第47位氨基酸，精氨酸被亮氨酸取代; 第51位氨基酸，被苯丙氨酸取代的酪氨酸; 第64位氨基酸，甘氨酸取代的甘氨酸; 第74位氨基酸，被甘氨酸取代的丙氨酸，第81位氨基酸，被异亮氨酸取代的苯丙氨酸，第86位氨基酸，被异亮氨酸取代的亮氨酸; 第87位氨基酸，用缬氨酸取代的苯丙氨酸; 第143位氨基酸，被甘氨酸取代的谷氨酸; 第188位氨基酸，用谷氨酰胺取代的亮氨酸; 和用缬氨酸取代的第267位氨基酸谷氨酸。