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    • 4. 发明授权
    • 산화적 안정성이 증대된 엔-카바모일라아제 변이효소
    • 산화적안정성이증대된엔 - 카바모일라아제변이효소
    • KR100464780B1
    • 2005-01-06
    • KR1020020014243
    • 2002-03-16
    • 한국과학기술원
    • 김학성오기훈남성훈
    • C12N15/55
    • 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.
    • 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。
    • 6. 发明公开
    • 기능요소의 동시 삽입에 의한 신기능을 갖는 단백질을제조하는 방법
    • 通过同时进行功能元素的具有新功能的蛋白质的制备方法
    • KR1020070058732A
    • 2007-06-11
    • KR1020050117353
    • 2005-12-05
    • 한국과학기술원
    • 김학성박희성남성훈김진현
    • C07K1/36
    • C12N15/1027C12N9/78C12N9/88C12N15/1044
    • A method for preparing a protein having new functions is provided to be widely utilized for developing protein for medicines and making industrial enzymes in the field of biotechnology. The method comprises the steps of: (a) designing function elements required for new functions which are desired to be provided to pre-existing protein scaffold; (b) simultaneously inserting at least two gene fragments corresponding to the designed function elements into a protein scaffold gene; and (c) selecting mutant having the new function from a library of mutants in which the new mutant genes are inserted and improving and adjusting the function of the selected mutant using a directed evolution technique such as an error prone-PCR and a DNA shuffling. The mutant protein evMBL8 of SEQ ID : NO. 29, in which function elements of a metallo beta-lactamase are introduced into a glyoxalase scaffold, is prepared by the method and deposited as a deposition number of KCTC 10877BP.
    • 提供一种制备具有新功能的蛋白质的方法,被广泛用于开发生物技术领域的药物蛋白质和制造工业酶。 该方法包括以下步骤:(a)设计期望提供给预先存在的蛋白质支架的新功能所需的功能元件; (b)同时将至少两个对应于所设计的功能元件的基因片段插入到蛋白质支架基因中; 和(c)从其中插入新的突变基因的突变体的文库中选择具有新功能的突变体,并使用定向进化技术(例如易错PCR和DNA改组)改进和调整所选突变体的功能。 SEQ ID NO:1的突变蛋白evMBL8。 29,其中将金属β-内酰胺酶的功能元件引入乙二醛酶支架中,通过该方法制备并沉积为KCTC 10877BP的沉积数。
    • 10. 发明公开
    • 산화적 안정성이 증대된 엔-카바모일라아제 변이효소
    • 具有改善的酸稳定性的杂化N-羧酸酶
    • KR1020030075098A
    • 2003-09-22
    • KR1020020014243
    • 2002-03-16
    • 한국과학기술원
    • 김학성오기훈남성훈
    • C12N15/55
    • C12N9/80C12N9/78C12N15/70C12P13/06C12P13/08C12Q1/34C12Q2334/00C12Y305/01077C12Y305/01087C12Y305/05001
    • 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.
    • 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。