公开(公告)号：KR100464780B1

公开(公告)日：2005-01-06

申请号：KR1020020014243

申请日：2002-03-16

Applicant: 한국과학기술원

Inventor： 김학성 ,  오기훈 ,  남성훈

IPC: C12N15/55

Abstract: PURPOSE: Mutated N-carbamoylase with improved acid-stability is provided, thereby effectively and cheaply producing synthetic D-amino acids without addition of DTT(dithiothreitol). CONSTITUTION: A gene encoding mutated N-carbamoylase has the nucleotide sequence of SEQ ID NO: 4, 12, 13 or 14, wherein the mutated N-carbamoylase is NC23, Gln23Leu, Val40AAla or Gly75Ser, respectively. A recombinant plasmid pNC23 contains the gene of SEQ ID NO: 4. A transformed Escherichia coli NC23(JM109/pTrc99a/NC23)(KCTC 10164BP) is produced by transforming with the recombinant plasmid pNC23. A method for producing the mutated N-carbamoylase comprises culturing the transformed Escherichia coli NC23(JM109/pTrc99a/NC23)(KCTC 10164BP). A method for producing synthetic D-amino acid comprises reacting the mutated N-carbamoylase with N-carbamoyl-D-amino acid, wherein the N-carbamoyl-D-amino acid is N-carbamoyl-D-para-hydroxyphenylglycine, N-carbamoyl-D-valine or N-carbamoyl-D-serine; and the synthetic D-amino acid is D-para-hydroxyphenylglycine, D-valine or D-serine.

Abstract translation: 目的：提供具有改善的酸稳定性的突变N-氨基甲酰基酶，由此有效且廉价地生产合成D-氨基酸而不添加DTT（二硫苏糖醇）。 构成：编码突变N-氨基甲酰基酶的基因具有SEQ ID NO：4,12,13或14的核苷酸序列，其中突变的N-氨基甲酰基酶分别是NC23，Gln23Le​​u，Val40AAla或Gly75Ser。 重组质粒pNC23含有SEQ ID NO：4的基因。用重组质粒pNC23转化产生转化的大肠杆菌NC23（JM109 / pTrc99a / NC23）（KCTC 10164BP）。 产生突变的N-氨基甲酰基酶的方法包括培养转化的大肠杆菌NC23（JM109 / pTrc99a / NC23）（KCTC 10164BP）。 生产合成D-氨基酸的方法包括使突变的N-氨基甲酰基酶与N-氨基甲酰基-D-氨基酸反应，其中N-氨基甲酰基-D-氨基酸是N-氨基甲酰基-D-对羟基苯基甘氨酸，N-氨基甲酰基 -D-缬氨酸或N-氨基甲酰基-D-丝氨酸; 并且合成的D-氨基酸是D-对羟基苯基甘氨酸，D-缬氨酸或D-丝氨酸。

公开(公告)号：KR1020030075098A

公开(公告)日：2003-09-22

申请号：KR1020020014243

申请日：2002-03-16

Applicant: 한국과학기술원

Inventor： 김학성 ,  오기훈 ,  남성훈

IPC: C12N15/55

CPC classification number: C12N9/80 ,  C12N9/78 ,  C12N15/70 ,  C12P13/06 ,  C12P13/08 ,  C12Q1/34 ,  C12Q2334/00 ,  C12Y305/01077 ,  C12Y305/01087 ,  C12Y305/05001

Abstract: PURPOSE: Mutated N-carbamoylase with improved acid-stability is provided, thereby effectively and cheaply producing synthetic D-amino acids without addition of DTT(dithiothreitol). CONSTITUTION: A gene encoding mutated N-carbamoylase has the nucleotide sequence of SEQ ID NO: 4, 12, 13 or 14, wherein the mutated N-carbamoylase is NC23, Gln23Leu, Val40AAla or Gly75Ser, respectively. A recombinant plasmid pNC23 contains the gene of SEQ ID NO: 4. A transformed Escherichia coli NC23(JM109/pTrc99a/NC23)(KCTC 10164BP) is produced by transforming with the recombinant plasmid pNC23. A method for producing the mutated N-carbamoylase comprises culturing the transformed Escherichia coli NC23(JM109/pTrc99a/NC23)(KCTC 10164BP). A method for producing synthetic D-amino acid comprises reacting the mutated N-carbamoylase with N-carbamoyl-D-amino acid, wherein the N-carbamoyl-D-amino acid is N-carbamoyl-D-para-hydroxyphenylglycine, N-carbamoyl-D-valine or N-carbamoyl-D-serine; and the synthetic D-amino acid is D-para-hydroxyphenylglycine, D-valine or D-serine.

Abstract translation: 目的：提供具有改善的酸稳定性的突变的N-氨基甲酰基酶，从而有效且廉价地生产合成的D-氨基酸而不添加DTT（二硫苏糖醇）。 构成：编码突变的N-氨基甲酰基酶的基因具有SEQ ID NO：4,12,13或14的核苷酸序列，其中突变的N-氨基甲酰基酶分别为NC23，Gln23Le​​u，Val40AAla或Gly75Ser。 重组质粒pNC23含有SEQ ID NO：4的基因。通过用重组质粒pNC23转化产生转化的大肠杆菌NC23（JM109 / pTrc99a / NC23）（KCTC 10164BP）。 制备突变的N-氨甲酰基酶的方法包括培养转化的大肠杆菌NC23（JM109 / pTrc99a / NC23）（KCTC10164BP）。 制备合成的D-氨基酸的方法包括使突变的N-氨基甲酰基酶与N-氨基甲酰基-D-氨基酸反应，其中N-氨基甲酰基-D-氨基酸是N-氨基甲酰基-D-对羟基苯基甘氨酸，N-氨基甲酰基 D-缬氨酸或N-氨基甲酰-D-丝氨酸; 并且合成的D-氨基酸是D-对羟基苯基甘氨酸，D-缬氨酸或D-丝氨酸。