Bacterial cell-surface displaying of thermo-tolerant glutamate dehydrogenase and its application in L-glutamate assay | |
Song, Jianxia1,2,3,4; Liang, Bo3,4; Han, Dongfei3,4; Tang, Xiangjiang3,4; Lang, Qiaolin3,4; Feng, Ruirui1,2; Han, Lihui1,2; Liu, Aihua3,4 | |
2015-03-01 | |
发表期刊 | ENZYME AND MICROBIAL TECHNOLOGY |
卷号 | 70期号:1页码:72-78 |
摘要 | In this paper, glutamate dehydrogenase (Gldh) is reported to efficiently display on Escherichia coli cell surface by using N-terminal region of ice the nucleation protein as an anchoring motif. The presence of Gldh was confirmed by SDS-PAGE and enzyme activity assay. Gldh was detected mainly in the outer membrane fraction, suggesting that the Gldh was displayed on the bacterial cell surface. The optimal temperature and pH for the bacteria cell-surface displayed Gldh (bacteria-Gldh) were 70 degrees C and 9.0, respectively. Additionally, the fusion protein retained almost 100% of its initial enzymatic activity after 1 month incubation at 4 degrees C. Transition metal ions could inhibit the enzyme activity to different extents, while common anions had little adverse effect on enzyme activity. Importantly, the displayed Gldh is most specific to L-glutamate reported so far. The bacterial Gldh was enabled to catalyze oxidization of L-glutamate with NADP(+) as cofactor, and the resultant NADPH can be detected spectrometrically at 340 nm. The bacterial-Gldh based L-glutamate assay was established, where the absorbance at 340 nm increased linearly with the increasing L-glutamate concentration within the range of 10-400 mu M. Further, the proposed approach was successfully applied to measure L-glutamate in real samples. (C) 2014 Elsevier Inc. All rights reserved. |
文章类型 | Article |
关键词 | Bacterial Surface Display L-glutamate Thermo-tolerant Glutamate Dehydrogenase Enzyme Inhibition L-glutamate Detection |
WOS标题词 | Science & Technology ; Life Sciences & Biomedicine |
DOI | 10.1016/j.enzmictec.2014.12.002 |
关键词[WOS] | ICE-NUCLEATION PROTEIN ; ESCHERICHIA-COLI ; WHOLE-CELL ; GLUCOSE-DEHYDROGENASE ; ANCHORING MOTIF ; D-XYLOSE ; ORGANOPHOSPHORUS HYDROLASE ; CEREBROSPINAL-FLUID ; MODIFIED-ELECTRODE ; ORGANIC-SOLVENTS |
收录类别 | SCI |
语种 | 英语 |
WOS研究方向 | Biotechnology & Applied Microbiology |
WOS类目 | Biotechnology & Applied Microbiology |
WOS记录号 | WOS:000350094500010 |
引用统计 | |
文献类型 | 期刊论文 |
条目标识符 | http://ir.qibebt.ac.cn/handle/337004/6091 |
专题 | 生物传感技术团队(过去) |
作者单位 | 1.Ocean Univ China, Minist Educ, Key Lab Marine Chem Theory & Technol, Qingdao 266100, Peoples R China 2.Ocean Univ China, Coll Chem & Chem Engn, Qingdao 266100, Peoples R China 3.Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, Lab Biosensing, Qingdao 266101, Peoples R China 4.Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, Key Lab Biofuels, Qingdao 266101, Peoples R China |
推荐引用方式 GB/T 7714 | Song, Jianxia,Liang, Bo,Han, Dongfei,et al. Bacterial cell-surface displaying of thermo-tolerant glutamate dehydrogenase and its application in L-glutamate assay[J]. ENZYME AND MICROBIAL TECHNOLOGY,2015,70(1):72-78. |
APA | Song, Jianxia.,Liang, Bo.,Han, Dongfei.,Tang, Xiangjiang.,Lang, Qiaolin.,...&Liu, Aihua.(2015).Bacterial cell-surface displaying of thermo-tolerant glutamate dehydrogenase and its application in L-glutamate assay.ENZYME AND MICROBIAL TECHNOLOGY,70(1),72-78. |
MLA | Song, Jianxia,et al."Bacterial cell-surface displaying of thermo-tolerant glutamate dehydrogenase and its application in L-glutamate assay".ENZYME AND MICROBIAL TECHNOLOGY 70.1(2015):72-78. |
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