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    • 3. 发明公开
    • 바실러스 속 균주 유래의 폴리감마글루탐산 합성유전자를이용한 표면 발현용 벡터 및 이를 이용한 단백질의 미생물표면 발현 방법
    • 包含PGSBCA,编码多糖 - 谷氨酸合成酶的基因的表面表达载体,以及使用其表达微生物表面上的靶标蛋白的方法
    • KR1020040032824A
    • 2004-04-17
    • KR1020037016665
    • 2002-08-09
    • 한국생명공학연구원주식회사 바이오리더스(주)엠디랩
    • 성문희홍승표이종수정창민김철중부하령소다겐지아시우치마코토
    • C12N15/75
    • C12N9/1051C07K2319/02C12N9/1288C12Y204/01133C12Y207/08005
    • PURPOSE: A surface expression vector including pgsBCA, a gene encoding poly-gamma-glutamate synthetase, and a method for expressing a target protein on the surface of a microorganism using the same are provided. The surface expression vector can be effectively used for the stable and easily detachable surface expression of various exogenous proteins onto the cell surface regardless of the cell cycle. CONSTITUTION: Surface expression vectors for producing a protein on a microbial surface contains one or more than two genes selected among pgsB, pgsC and pgsA, encoding a poly-γ-glutamate synthetase complex, wherein the gene is derived from a Bacillus sp. strain producing poly-γ-glutamate synthetase; and the nucleotide sequences of pgsB, pgsC and pgsA genes are homologous to those of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, respectively, with an 80% homology. A method for expressing a target protein on the surface of a microorganism comprises the steps of: (a) constructing a recombinant expression vector by inserting a gene encoding the target protein into the surface expression vector; (b) transforming a Gram-negative host cell with the recombinant vector; and cultivating the transformed host cell and expressing the target protein on the surface of the host cell.
    • 目的:提供包含pgsBCA,编码聚-γ-谷氨酸合成酶的基因的表面表达载体和使用其的表达微生物的靶蛋白的方法。 无论细胞周期如何,表面表达载体都可以有效地用于将各种外源蛋白质稳定且容易地分离到细胞表面上的表面表达。 构成:用于在微生物表面上产生蛋白质的表面表达载体含有一个或多于两个选自pgsB,pgsC和pgsA的基因,编码聚-γ-谷氨酸合成酶复合物,其中该基因源自芽孢杆菌属。 生产聚-γ-谷氨酸合成酶的菌株; pgsB,pgsC和pgsA基因的核苷酸序列分别与SEQ ID NO:1,SEQ ID NO:2和SEQ ID NO:3的核苷酸序列同源,具有80%的同源性。 用于在微生物表面上表达靶蛋白的方法包括以下步骤:(a)通过将编码靶蛋白的基因插入表面表达载体来构建重组表达载体; (b)用重组载体转化革兰氏阴性宿主细胞; 并培养转化的宿主细胞并在宿主细胞的表面上表达靶蛋白。
    • 6. 发明公开
    • 고분자량의 폴리-감마-글루탐산을 생산하는 내염성 균주 바실러스 서브틸리스 청국장
    • 生产高分子量聚乳酸的BSA-4能够生产聚氨基酸的高分子量的HALOPHILE MICROORGANISM BACILLUS SUBTILIS BS-4
    • KR1020020079889A
    • 2002-10-19
    • KR1020027010976
    • 2001-08-11
    • 한국생명공학연구원주식회사 바이오리더스
    • 성문희백대헌이승구송재준홍승표최윤호한재숙아시우치마코토소다겐지부하령나유진
    • C12N1/20
    • C12R1/125C07K14/32
    • PURPOSE: Provided are halophile microorganism Bacillus subtilis BS-4 capable of producing high molecular weight of poly-γ-glutamic acid and a method for producing the high molecular weight of poly-γ-glutamic acid using the same, thereby producing the high molecular weight poly-γ-glutamic acid in higher yield. CONSTITUTION: The halophile microorganism Bacillus subtilis BS-4 (KCTC 0697BP) capable of producing high molecular weight of poly-γ-glutamic acid is isolated from 'Chug Kook Jang'(Fermented soybean paste) by the steps of: diluting 'Chug Kook Jang' in distilled water; heating the diluted solution at 60 deg. C for 20 minutes; plating the diluted solution on GS agar medium; culturing it at 37 deg. C for 3 days; isolating high viscosity of colonies; culturing the colonies in a broth at 37 deg. C for 20 hours; and selecting a strain showing the best growth activity. The method for producing the high molecular weight of poly-γ-glutamic acid comprises the steps of: (a) culturing the Bacillus subtilis BS-4(KCTC 0697BP) to obtain poly-γ-glutamic acid; and (b) removing polysaccharides from the poly-γ-glutamic acid; dissolving the precipitate of poly-γ -glutamic acid and adding a protease into the solution to decompose an extracellular protein; and dialyzing the solution to remove free glutamic acid and concentrating the solution.
    • 目的:提供能够生产高分子量聚-γ-谷氨酸的嗜碱性微生物枯草芽孢杆菌BS-4,以及使用其制备高分子量聚-γ-谷氨酸的方法,从而产生高分子量 聚-γ-谷氨酸。 构成:通过以下步骤从“Chug Kook Jang”(发酵大豆酱)分离能够产生高分子量聚-γ-谷氨酸的嗜碱性微生物枯草芽孢杆菌BS-4(KCTC 0697BP),稀释“Chug Kook Jang 在蒸馏水中 将稀释溶液加热至60℃。 C 20分钟; 将稀释的溶液电镀在GS琼脂培养基上; 在37度培养 C 3天; 分离菌落的高粘度; 在37℃的肉汤培养菌落。 C 20小时; 并选择显示最佳生长活性的菌株。 制备高分子量聚-γ-谷氨酸的方法包括以下步骤:(a)培养枯草芽孢杆菌BS-4(KCTC 0697BP)以获得聚-γ-谷氨酸; 和(b)从聚-γ-谷氨酸中除去多糖; 溶解聚-γ-谷氨酸的沉淀物,并向溶液中加入蛋白酶以分解细胞外蛋白质; 并透析溶液以除去游离谷氨酸并浓缩溶液。
    • 8. 发明公开
    • 고분자량의 폴리-감마-글루탐산을 생산하는 내염성 균주바실러스 서브틸리스 청국장
    • BACILLUS SUBTILISVAR。 CHUNGKOOKJANG(KTCT 0697BP)生产高分子量的多糖 - 谷氨酸
    • KR1020010078440A
    • 2001-08-21
    • KR1020010001481
    • 2001-01-11
    • 주식회사 바이오리더스가부시키 가이샤 파마푸즈
    • 성문희백대헌이승구송재준홍승표최윤호신선영한재숙아시우치마코토소다겐지
    • C12N1/20
    • C12R1/125C07K14/32
    • PURPOSE: Provided is salt tolerant strain, Bacillus subtilisvar. chungkookjang (KTCT 0697BP) isolated from Korean traditional fermented food, chungkookjang. And poly-gamma-glutamic acid is also provided which is edible, water-soluble, anionic, biodegradable and produced from the strain, and which is used in cosmetics, absorbent, biodegradable plastic materials. CONSTITUTION: A Bacillus subtilisvar. chungkookjang (KTCT 0697BP) is isolated from chungkookjang. A preparation method of poly-gamma-glutamic acid from the strain comprises the steps of: (a) inoculating and cultivating Bacillus subtilisvar. chungkookjang (KTCT 0697BP) in a medium to produce poly-gamma-glutamic acid and then, adding acids until PH becomes 2 to 4 , to obtain liquid that includes poly-gamma-glutamic acid; (b) removing polysaccharide from the resultant liquid, followed by extracting with solvent and centrifuging it to obtain precipitate of poly-gamma-glutamic acid; (c) dissolving the precipitant by adding distilled water, then degrading exogenous protein by proteinase; and (d) dialyzing it to remove free glutamic acid and concentrating it.
    • 目的:提供耐盐菌株,枯草芽胞杆菌菌。 chungkookjang(KTCT 0697BP)从韩国传统发酵食品,chungkookjang分离。 还提供了可从食品,水溶性,阴离子性,生物降解性和从菌株生产的聚γ-谷氨酸,并且用于化妆品,吸收性,可生物降解的塑料材料中。 构成:枯草芽孢杆菌。 chungkookjang(KTCT 0697BP)从chungkookjang隔离。 来自菌株的聚-γ-谷氨酸的制备方法包括以下步骤:(a)接种并培养枯草芽孢杆菌菌素。 (KTCT 0697BP)在培养基中产生聚-γ-谷氨酸,然后加入酸直到PH变为2〜4,得到包含聚-γ-谷氨酸的液体; (b)从所得液体中除去多糖,然后用溶剂萃取,离心分离得到聚-γ-谷氨酸沉淀物; (c)通过加入蒸馏水溶解沉淀剂,然后通过蛋白酶降解外源蛋白质; 和(d)透析以除去游离谷氨酸并浓缩。