Construction of a fusion enzyme system by gene splicing as a new molecular recognition element for a sequence biosensor
文献类型：期刊论文
| 作者 | Zhou, YF; Zhang, XE; Liu, H; Zhang, ZP; Zhang, CG; Cass, AEG; Zhang, XE, Chinese Acad Sci, Wuhan Inst Virol, Wuhan 430071, Peoples R China |
| 刊名 | BIOCONJUGATE CHEMISTRY
 |
| 出版日期 | 2001 |
| 卷号 | 12期号:6页码:924-931 |
| ISSN号 | 1043-1802 |
| 英文摘要 | A bifunctional fusion enzyme system constructed by gene splicing is proposed as a new model to develop sequence biosensors, taking maltose biosensor as an example. The cDNA fragment of Aspergillus niger glucoamylase (E.C 3.2.1.3, GA) was fused to the 3 end of Aspergillus niger glucose oxidase (E.C 1.1.3.4, GOD) gene with the insertion of a flexible linker peptide [-(Ser-Gly)(5)-] coding sequence. The fusion gene was cloned into the vector pPIC9 and expressed in Pichia pastoris GS115 under the control of the AOX1 promoter. It was found that, a bifunctional hybrid protein with a molecular weight of 430 kDa was secreted after induction with methanol. The fusion enzyme GOD-(Ser-Gly)(5)-GA (GLG) was purified using Q Sepharose Fast Flow ion-exchang chromatography. Kinetic analysis demonstrated that GLG retained the typical kinetic properties of both GA and GOD. After being immobilized on an aminosilanized glass slide through covalent bonding by glutaraldehyde, GLG showed much higher sequential catalytic efficiency than the mixture of separately expressed GA and GOD (GA/GOD). Maltose biosensors were fabricated with GLG and GA/GOD, respectively. The performance characteristics of the maltose biosensor with respect to reproducibility, signal level, and linearity were effectively improved by using the fusion enzyme. Our findings offer a basis for the development of other sequence biosensors. |
| 学科主题 | Biochemical Research Methods ; Biochemistry & Molecular Biology ; Chemistry, Multidisciplinary ; Chemistry, Organic |
| 语种 | 英语 |
| WOS记录号 | WOS:000172451200014 |
| 公开日期 | 2011-09-23 |
| 源URL | [http://210.72.129.5/handle/321005/55513]  |
| 专题 | 沈阳应用生态研究所_沈阳应用生态研究所 |
| 通讯作者 | Zhang, XE, Chinese Acad Sci, Wuhan Inst Virol, Wuhan 430071, Peoples R China |
| 推荐引用方式 GB/T 7714 | Zhou, YF,Zhang, XE,Liu, H,et al. Construction of a fusion enzyme system by gene splicing as a new molecular recognition element for a sequence biosensor[J]. BIOCONJUGATE CHEMISTRY,2001,12(6):924-931. |
| APA | Zhou, YF.,Zhang, XE.,Liu, H.,Zhang, ZP.,Zhang, CG.,...&Zhang, XE, Chinese Acad Sci, Wuhan Inst Virol, Wuhan 430071, Peoples R China.(2001).Construction of a fusion enzyme system by gene splicing as a new molecular recognition element for a sequence biosensor.BIOCONJUGATE CHEMISTRY,12(6),924-931. |
| MLA | Zhou, YF,et al."Construction of a fusion enzyme system by gene splicing as a new molecular recognition element for a sequence biosensor".BIOCONJUGATE CHEMISTRY 12.6(2001):924-931. |
入库方式： OAI收割
来源：沈阳应用生态研究所
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