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公开(公告)号：KR1020110060384A
公开(公告)日：2011-06-08
申请号：KR1020090116957
申请日：2009-11-30
Applicant: 한국화학연구원 , 충남대학교산학협력단
Abstract: PURPOSE: A Candida Antarctica-derived lipase B(CalB) protein variant is provided to increase enzyme activity and to be used in the industry. CONSTITUTION: A CalB protein variant is selected from a protein of amino acid sequences 2-8 in which 274-289th amino acid is partially substituted in Candida Antarctica-derived lipase B(CalB) protein. A gene encoding CalB protein variant sequence is denoted by sequence number selected from sequence numbers 10-16. A recombinant vector containing the gene is prepared by inserting the gen into pK7 vector. A transformed cell which produces CalB protein variant is obtained by transforming a host cell with the recombinant vector. The host cells are Pichia pastoris.
Abstract translation: 目的：提供来自南极假丝酵母的脂肪酶B（CalB）蛋白质变体以增加酶活性并用于该行业。 构成：CalB蛋白质变体选自氨基酸序列2-8的蛋白质，其中274-289位氨基酸被部分取代在南极假单胞菌来源的脂肪酶B（CalB）蛋白质中。 编码CalB蛋白变体序列的基因由选自序列号10-16的序列号表示。 通过将gen插入pK7载体中制备含有该基因的重组载体。 通过用重组载体转化宿主细胞获得产生CalB蛋白质变体的转化细胞。 宿主细胞是巴斯德毕赤酵母。
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公开(公告)号：KR101176802B1
公开(公告)日：2012-08-27
申请号：KR1020090116957
申请日：2009-11-30
Applicant: 한국화학연구원 , 충남대학교산학협력단
Abstract: 본 발명은 캔디다 안타르크티카(
Candida antarctica ) 유래의 리파제 B(CalB) 단백질 변이체에 관한 것으로서, 서열번호 1의 아미노산 서열로 표시되는 CalB 단백질의 274 위치 내지 289 위치의 아미노산 중 일부가 치환된, 서열번호 2 내지 8의 아미노산 서열로 표시되는 단백질로 이루어진 군에서 선택되는 본 발명에 따른 CalB 단백질 변이체는, 기존의 CalB 단백질보다 최대 35 배 증가된 효소 활성을 가지므로 리파제를 이용하는 산업에 유용하게 사용될 수 있다.
생명공학, 효소, 변이, 단백질, 활성-
公开(公告)号：KR100733712B1
公开(公告)日：2007-06-29
申请号：KR1020060015605
申请日：2006-02-17
Applicant: 충남대학교산학협력단
Abstract: A preparation method of a cell extract useful as catalyst for synthesis of a cell-free protein and the cell-free protein synthesis method by using the same extract are provided to improve economical efficiency and productivity of the cell-free protein synthesis by simplifying the preparation process of the cell extract through high-speed centrifugation. The preparation method of the cell extract useful as catalyst for synthesis of the cell-free protein comprises the steps of: destroying the cells cultured in the medium to prepare destroyed cell solution containing cellular organelles and factors required for synthesis of a desired protein; centrifuging the destroyed cell solution at 12,000 to 30,000Xg and collecting the supernatant to prepare the cell extract; and adding the cell extract into a reaction medium containing amino acid mixture, protein synthesis energy source, gene information source and buffer solution, wherein the cell is Escherichia coli, Bacillus subtilis, wheat germ, rice germ, barley germ, CHO(Chinese hamster ovary) cell, hybridoma cell or reticulocyte; the protein synthesis energy source is ATP(adenine triphosphate), CTP(cytidine triphosphate), GTP(guanosine triphosphate), TTP(thymine triphosphate) or UTP(uridine triphosphate); and the gene information source is DNA or mRNA(messenger RNA) encoding the target protein.
Abstract translation: 提供一种可用作合成无细胞蛋白质的催化剂的细胞提取物的制备方法和使用相同提取物的无细胞蛋白质合成方法，以通过简化制备来提高无细胞蛋白质合成的经济效率和生产率 通过高速离心进行细胞提取物的过程。 用作合成无细胞蛋白质的催化剂的细胞提取物的制备方法包括以下步骤：破坏在培养基中培养的细胞以制备含有细胞器的破坏细胞溶液和合成期望蛋白质所需的因子; 以12,000至30,000Xg离心破坏的细胞溶液并收集上清液以制备细胞提取物; 将细胞提取物加入到含有氨基酸混合物，蛋白质合成能源，基因信息来源和缓冲溶液的反应介质中，其中细胞为大肠杆菌，枯草芽孢杆菌，小麦胚芽，米胚芽，大麦胚芽，CHO（中国仓鼠卵巢 ）细胞，杂交瘤细胞或网织红细胞; 蛋白质合成能源是ATP（三磷酸腺嘌呤），CTP（三磷酸胞苷），GTP（鸟苷三磷酸），TTP（三磷酸胸腺嘧啶）或UTP（尿苷三磷酸）; 而基因信息来源是编码靶蛋白的DNA或mRNA（信使RNA）。
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