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公开(公告)号：KR1020080078777A
公开(公告)日：2008-08-28
申请号：KR1020070112892
申请日：2007-11-06
Applicant: 서강대학교산학협력단 , 국민대학교산학협력단
IPC: C12Q1/18
CPC classification number: C12Q1/025 , G01N2333/90283
Abstract: A method for screening an antioxidant using mutant bacteria and chlorophyllide is provided to screen a large amount of the antioxidant easily by monitoring the growth state of the mutant bacteria and be useful for selective screening of an amphiphilic antioxidant, thereby applying the effective antioxidant with little toxicity as an additive of various foods and a cosmetic product. A method for screening an antioxidant comprises the steps of: (a) culturing mutant bacteria, which is transformed by including mutation in a DNA sequence encoding superoxide dismutase, thereby inhibiting the activity of the superoxide dismutase and expressing chlorophyllide reductase capable of inducing generation of superoxide anion in the presence of oxygen, in a plate; (b) locating a plurality of filter discs where an antioxidant candidate material to be screened and chlorophyllide are added on the plate; and (c) selecting the filter disc where the growth of the mutant bacteria is observed, wherein the bacteria to be mutated is Rhodobacter sphaeroides, Rhodospirillum rubrum, Rhodopseudomonas palustris, Allochromatium vinosum, Chlorobium tepidum or Chloroflexus aurantiacus. Further, an adding weight of chlorophyllide is 0.1 to 100 mM.
Abstract translation: 提供使用突变型细菌和叶绿素来筛选抗氧化剂的方法，通过监测突变细菌的生长状态容易地筛选大量的抗氧化剂，并且可用于选择性筛选两亲性抗氧化剂，从而将有效的抗氧化剂施用少量毒性 作为各种食品和化妆品的添加剂。 筛选抗氧化剂的方法包括以下步骤：（a）通过在编码超氧化物歧化酶的DNA序列中包含突变而转化突变细菌，从而抑制超氧化物歧化酶的活性并表达能诱导产生超氧化物的叶绿素还原酶 阴离子在氧的存在下，在板中; （b）定位多个滤盘，其中待筛选的抗氧化剂候选材料和叶绿素被添加到板上; （c）选择观察到突变细菌的生长的滤膜，其中待突变的细菌是球形红细菌，红色红孢菌，红假罗毕假单胞菌，紫苏胶，氧化铁或绿脓杆菌。 此外，叶绿素的添加重量为0.1〜100mM。
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公开(公告)号：KR1020080076502A
公开(公告)日：2008-08-20
申请号：KR1020070016514
申请日：2007-02-16
Applicant: 서강대학교산학협력단
Abstract: A reagent generating superoxide anion is provided to improve production yield and stability of superoxide anion by inhibiting generation of hydrogen peroxide, so that the reagent is useful for screening superoxide anion-removing materials and for analyzing activity of superoxide dismutase. A reagent generating superoxide anion contains BchZ protein generating superoxide anion with generation of hydrogen peroxide, represented by SEQ ID NO:2, and derived from photosynthetic bacteria. A superoxide anion-removing material is screened by contacting a candidate superoxide anion-removing material and luminal derivatives with the superoxide anion-generating reagent, and measuring luminescence intensity. The activity of superoxide dismutase is analyzed by adding cytochrome C or NBT(nitro blue tetrazolium) into the superoxide anion-generating reagent to induce reduction of cytochrome C or NBT, and measuring absorbance changes of the reaction product. Further, a hemin is connected with the BchZ protein.
Abstract translation: 提供产生超氧阴离子的试剂，以通过抑制过氧化氢的产生来提高超氧阴离子的产率和稳定性，使得该试剂可用于筛选超氧阴离子去除材料和分析超氧化物歧化酶的活性。 产生超氧化物阴离子的试剂含有产生超氧阴离子的BchZ蛋白，产生由SEQ ID NO：2表示的过氧化氢，并衍生自光合细菌。 通过使候选超氧阴离子去除材料和管腔衍生物与超氧阴离子产生试剂接触并测量发光强度来筛选超氧阴离子去除材料。 通过将细胞色素C或NBT（硝基蓝四唑）加入超氧阴离子产生试剂中来分析超氧化物歧化酶的活性，以诱导细胞色素C或NBT的还原，并测量反应产物的吸光度变化。 此外，血红素与BchZ蛋白相关。
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公开(公告)号：KR100814502B1
公开(公告)日：2008-03-18
申请号：KR1020070015913
申请日：2007-02-15
Applicant: 서강대학교산학협력단
CPC classification number: C12N15/63 , C12N15/113
Abstract: A method for screening genes regulating oxidative stress is provided to investigate harmfulness of active oxygen by directly controlling the generation of superoxide active oxygen in specific cell organelles and search genes and physiologically active materials inhibiting outbreak of various diseases generated from various harmfulness of the active oxygen and harmful effect of the active oxygen. A method for screening genes regulating oxidative stress with high throughput comprises the steps of: (a) preparing a recombinant expression vector to express gene coding polymer proteins inducing generation of superoxide anions; (b) transfecting the expression vector into a cell to clone a gene expression single cell line; and (c) after putting the expression vector in the cell line and adding chlorophyllide thereto, measuring apoptosis caused by active oxygen, wherein the gene coding the polymer protein includes bchX described as SEQ ID : NO. 1, bchY described as SEQ ID : NO. 2 and bchZ described as SEQ ID : NO. 3.
Abstract translation: 提供了筛选调节氧化应激基因的方法，通过直接控制特定细胞器中的超氧化物活性氧的产生和搜索基因以及生理活性物质抑制由活性氧的各种有害性引起的各种疾病的爆发，来研究活性氧的危害， 活性氧的有害影响。 用于筛选以高通量调节氧化应激的基因的方法包括以下步骤：（a）制备重组表达载体以表达导致产生超氧化物阴离子的编码聚合物蛋白的基因; （b）将表达载体转染入细胞以克隆基因表达单细胞系; 和（c）在将表达载体置于细胞系中并加入叶绿素后，测定由活性氧引起的凋亡，其中编码聚合蛋白的基因包括如SEQ ID NO：1所示的bchX。 1，bchY描述为SEQ ID NO： 2和bchZ如SEQ ID NO： 3。
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公开(公告)号：KR100861126B1
公开(公告)日：2008-09-30
申请号：KR1020070016514
申请日：2007-02-16
Applicant: 서강대학교산학협력단
Abstract: 본 발명은 슈퍼옥시드 음이온 생성 시약 및 이의 용도에 관한 것으로서, 더욱 상세하게는 산소 조건 하에서 과산화수소를 생성하지 않으면서 슈퍼옥시드 음이온을 생성하는 활성을 가진 로도박터(
Rhodobacter )를 포함하는 광합성 세균 유래 단백질을 이용한 슈퍼옥시드 음이온 생성 시약으로 사용하여, 직접 슈퍼옥시드 음이온 제거물질을 검색하거나, 또는 시토크롬 C나 NBT(nitro blue tetrazolium)를 이용하여 간접적으로, 생성된 슈퍼옥시드 음이온에 의하여 시토크롬 C나 NBT가 환원된 것을 흡광도의 변화를 측정함으로써, 시료에 있는 슈퍼옥시드 디스뮤타제 양에 비례하여 슈퍼옥시드 생성 시약에 의해 생성된 슈퍼옥시드 음이온의 분해율의 상관관계를 이용하여 슈퍼옥시드 디스뮤타제의 활성 측정하는 방법에 관한 것이다.
슈퍼옥시드 음이온 생성 시약, 항산화활성 검색, 슈퍼옥시드 디스뮤타제 활성 측정
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