公开(公告)号：KR100818611B1

公开(公告)日：2008-04-01

申请号：KR1020060078204

申请日：2006-08-18

Applicant: 재단법인서울대학교산학협력재단

Inventor： 김병기 ,  김주한 ,  경도현 ,  윤형돈

IPC: C12N1/20 ,  C12P21/00

Abstract: 본 발명은 토양으로부터 방향족 베타아미노산을 유일한 질소원으로 포함하는 최소배지를 이용하여 분리한 광학선택성이 높은 베타트랜스아미나제 활성을 갖는 신규한 미생물인 메조리조비움 sp. LUK (Mesorhizobium sp. LUK)와 상기 분리된 미생물로부터 분리 정제한 베타트랜스아미나제를 암호화하는 유전자에 관한 것이다.
또한, 본 발명은 상기 분리된 토양 미생물 또는 이의 세포 파쇄물로부터 수득된 방향족 베타트랜스아미나제 추출물을 생촉매로 이용하여 광학적으로 순수한 방향족 L형- 또는 D형-베타아미노기 화합물을 생산하는 방법에 관한 것이다.
베타아미노산, 트랜스아미나제, 베타트랜스아미나제

公开(公告)号：KR1020080016287A

公开(公告)日：2008-02-21

申请号：KR1020060078204

申请日：2006-08-18

Applicant: 재단법인서울대학교산학협력재단

Inventor： 김병기 ,  김주한 ,  경도현 ,  윤형돈

IPC: C12N1/20 ,  C12P21/00

Abstract: A novel microorganism primarily isolated from soil using a minimum culture medium including a beta amino acid as a single nitrogen source is provided to show the high optical selectivity and transaminase activity on a D-type beta amino acid. And a method for producing an L-type or a D-type beta amino acid is provided to be able to obtain the enantiomerically pure product using the same microorganism. A Mesorhizobium sp. LUK is deposited as a deposition no. KCCM-10752P. An L-type beta amino acid is produced at a temperature of 20-70 deg.C under pH of 5-10 by using the Mesorhizobium sp. LUK or a cell extract thereof as a bio-catalyst through optical resolution. A D-type beta amino acid is produced at a temperature of 20-70 deg.C under pH of 5-10 by using the Mesorhizobium sp. LUK or the cell extract thereof as a bio-catalyst through asymmetric synthesis. A transformed host cell is transformed by a recombinant DNA vector including a beta-transaminase gene encoding an amino acid sequence of SEQ ID : NO. 3 or a gene having more than 97% homology to the sequence of SEQ ID : NO. 3 and encoding a protein with the beta-transaminase activity. Further, an amino accepter for the optical resolution is selected from keto acid group.

Abstract translation: 提供了使用包含β氨基酸作为单一氮源的最小培养基从土壤中主要分离的新型微生物，以显示对D型β氨基酸的高光学选择性和转氨酶活性。 并且提供了用于生产L型或D型β氨基酸的方法以能够使用相同的微生物获得对映异构体纯的产物。 中生根瘤菌 LUK作为沉积物进行沉积。 KCCM-10752P。 通过使用中生根瘤菌属（Mesorhizobium sp。），在pH 5-10的温度下，在20-70℃的温度下产生L型β-氨基酸。 LUK或其细胞提取物作为生物催化剂通过光学拆分。 通过使用中生根瘤菌属（Mesorhizobium sp。），在pH 5-10的温度下，在20-70℃的温度下产生D型β氨基酸。 LUK或其细胞提取物作为通过不对称合成的生物催化剂。 通过重组DNA载体转化转化的宿主细胞，所述重组DNA载体包含编码SEQ ID NO：1的氨基酸序列的β-转氨酶基因。 3或与SEQ ID NO：3的序列具有超过97％同源性的基因。 3并编码具有β-转氨酶活性的蛋白质。 此外，用于光学拆分的氨基取代基选自酮酸基团。

公开(公告)号：KR1020020043349A

公开(公告)日：2002-06-10

申请号：KR1020000072824

申请日：2000-12-04

Applicant: 재단법인서울대학교산학협력재단

Inventor： 김병기 ,  김주한

IPC: C12N11/04

CPC classification number: C12N11/04

Abstract: PURPOSE: Provided are an immobilized phospholipase A-2 and a method for hydrolysis of phospholipid or phospholipid-containing lecithin by using it. Thereby, enzymes can be reused and consequently, the costs for hydrolysis of phospholipid can be decreased. CONSTITUTION: The phospholipase A-2 is immobilized in a calcium alginate bead and is coated with a gelating agent, wherein the gelating agent is selected from the group consisting of tetraalkoxysilane wherein the alkoxy is selected from methoxy, ethoxy, propoxy, isopropoxy, and butoxy; and aminoalkoxy trialkoxysilane and carboxyalkyltrialkoxysilane, wherein the alkyl is C1 to C50 linear saturated carbohydrate and the alkoxy is the same above. The method for hydrolysis of phospholipid or phospholipid-containing lecithin comprises dissolving phospholipid or phospholipid-containing lecithin in a mixed solution of buffer and organic solvent using the immobilized phospholipase A-2, in which divalent cationic ions such as calcium, magnesium or iron is added into the solution; the reaction temperature is 30 to 50 deg. C; the hydrogen concentration of the solution is pH 8 to 10; and the organic solvent is selected from methanol, ethanol, butanol, 2-butanol, t-butanol, propanol, isopropanol or a combination thereof.

Abstract translation: 目的：提供一种固定化磷脂酶A-2和通过使用它来水解磷脂或磷脂卵磷脂的方法。 因此，酶可以重复使用，因此可以降低磷脂水解的成本。 构成：将磷脂酶A-2固定在海藻酸钙珠中，并用凝胶剂包衣，其中凝胶剂选自四烷氧基硅烷，其中烷氧基选自甲氧基，乙氧基，丙氧基，异丙氧基和丁氧基 ; 和氨基烷氧基三烷氧基硅烷和羧烷基三烷氧基硅烷，其中烷基是C1至C50线性饱和碳水化合物，并且烷氧基与上述相同。 含磷脂或含磷脂的卵磷脂的水解方法包括使用固定化磷脂酶A-2将含磷脂或磷脂的卵磷脂溶解在缓冲液和有机溶剂的混合溶液中，其中添加了诸如钙，镁或铁的二价阳离子离子 进入解决方案; 反应温度为30〜50℃。 C; 溶液的氢浓度为8〜10; 有机溶剂选自甲醇，乙醇，丁醇，2-丁醇，叔丁醇，丙醇，异丙醇或其组合。