公开(公告)号：KR100818982B1

公开(公告)日：2008-04-04

申请号：KR1020060114786

申请日：2006-11-20

Applicant: 주식회사농심 ,  재단법인서울대학교산학협력재단

Inventor： 이형주 ,  안창원 ,  신동석 ,  정도원 ,  한기선

IPC: C12N15/64 ,  C12N15/63

Abstract: A method for mass-producing a recombinant antihypertensive peptide is provided to be usefully used for producing a peptide having low molecular weight at low cost with high efficiency and use the antihypertensive peptide for a recombinant expression system for foods. A method for preparing a recombinant pEUH40 having a cleavage map depicted in Fig. 2 comprises the steps of: (a) inserting a gene of SEQ ID : NO. 1 into a cloning vector pUC18; (b) after self-ligating the gene of SEQ ID : NO. 1, inserting the self-ligated gene into the recombinant vector obtained from the step(a) to transform E. coli DH5alpha; and (c) after inserting the vector obtained from the step(b) into a vector pET29b, fusing a ubiquitin gene thereinto. A method for mass-producing a recombinant antihypertensive peptide His-His-Leu comprises the steps of: (a) culturing 40 copies of His-His-Leu sequences using a transgenic E.coli KCTC 11004BP including the recombinant pEUH40 plasmid; (b) after centrifuging the culture material, eluting cells therefrom and extracting and purifying a cell eluent with cation exchange chromatography; and (c) after hydrolyzing the purified material with leucine aminopeptidase, isolating His-His-Leu from the hydrolysate through HPLC.

Abstract translation: 提供了大规模生产重组抗高血压肽的方法，可有效地用于高效率地低成本生产低分子量的肽，并使用抗高血压肽作为食品的重组表达系统。 制备具有图1所示切割图的重组pEUH40的方法。 2包括以下步骤：（a）插入SEQ ID NO： 1转化成克隆载体pUC18; （b）在自身连接SEQ ID NO： 1，将自连接基因插入从步骤（a）获得的重组载体中，以转化大肠杆菌DH5α; 和（c）将从步骤（b）获得的载体插入载体pET29b之后，将泛素基因融合在其中。 用于大规模生产重组抗高血压肽His-His-Leu的方法包括以下步骤：（a）使用包含重组pEUH40质粒的转基因大肠杆菌KCTC 11004BP培养40个拷贝的His-His-Leu序列; （b）离心培养物后，从其中洗脱细胞，用阳离子交换色谱提取和纯化细胞洗脱液; 和（c）在用亮氨酸氨基肽酶水解纯化的物质后，通过HPLC从水解产物中分离出His-His-Leu。

公开(公告)号：KR1020080061611A

公开(公告)日：2008-07-03

申请号：KR1020060136535

申请日：2006-12-28

Applicant: 주식회사농심

Inventor： 송인상 ,  박창신 ,  안창원 ,  강주희 ,  이호현 ,  한기선

IPC: C12P21/06

CPC classification number: C12P21/06 ,  C12N1/20 ,  C12N9/0083 ,  C12Y114/99003

Abstract: A soybean protein hydrolysate is provided to induce expression of heme oxygenase-1(HO-1), protect cells by effects of HO-1 including anti-inflammation, cell proliferation and apoptosis inhibition, and protect and treat vascular damage shown in obesity, hypertension, impotence, dysuria, diabetes, arthritis and aging. A soybean protein hydrolysate for increasing expression of heme oxygenase-1 is prepared by hydrolyzing soybeans with endopeptidase and exopeptidase, or fermenting soybeans with a microorganism selected from Bacillus sp., Leuconostoc sp., Lactobacillus sp. and Aspergillus sp., separating the hydrolyzed or fermented solution into a solid phase and a liquid phase, and filtering and powdering the liquid phase, wherein the soybean is black soybean; the microorganism is Bacillus subtilis(KCTC 11029BP); and the soybean protein hydrolysate activate Co-cGMP(cyclic guanosine mono-phosphate) pathway.

Abstract translation: 提供大豆蛋白水解物以诱导血红素加氧酶-1（HO-1）的表达，通过HO-1的作用保护细胞，包括抗炎，细胞增殖和凋亡抑制，并保护和治疗肥胖，高血压中显示的血管损伤 ，无能，排尿困难，糖尿病，关节炎和衰老。 通过用内肽酶和外肽酶水解大豆，或用选自芽孢杆菌（Bacillus sp。），明串珠菌（Leuconostoc sp。），乳杆菌属（Lactobacillus sp。））的微生物发酵大豆来制备用于增加血红素加氧酶-1表达的大豆蛋白水解产物。 和曲霉菌，将水解或发酵的溶液分离成固相和液相，过滤和粉化液相，其中大豆是黑豆; 该微生物是枯草芽孢杆菌（KCTC 11029BP）; 大豆蛋白水解物活化Co-cGMP（环鸟苷酸一磷酸）途径。