Directed evolution of the PobR allosteric transcription factor to generate a biosensor for 4-hydroxymandelic acid
Hydroxy-mandelic acid (HMA) is widely applied in pharmaceuticals, food and cosmetics. In this study, we aimed to develop an allosteric transcription factors (aTFs) based biosensor for HMA. PobR, an aTF for HMA analog 4-hydroxybenzoic acid, was used to alter its selectivity and create novel aTFs resp...
Main Authors: | , , , , , , , , , , , , |
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Format: | Article |
Language: | English |
Published: |
Springer Science and Business Media B.V.
2022
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Subjects: | |
Online Access: | View Fulltext in Publisher |
LEADER | 02398nam a2200505Ia 4500 | ||
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001 | 10.1007-s11274-022-03286-5 | ||
008 | 220706s2022 CNT 000 0 und d | ||
020 | |a 09593993 (ISSN) | ||
245 | 1 | 0 | |a Directed evolution of the PobR allosteric transcription factor to generate a biosensor for 4-hydroxymandelic acid |
260 | 0 | |b Springer Science and Business Media B.V. |c 2022 | |
856 | |z View Fulltext in Publisher |u https://doi.org/10.1007/s11274-022-03286-5 | ||
520 | 3 | |a Hydroxy-mandelic acid (HMA) is widely applied in pharmaceuticals, food and cosmetics. In this study, we aimed to develop an allosteric transcription factors (aTFs) based biosensor for HMA. PobR, an aTF for HMA analog 4-hydroxybenzoic acid, was used to alter its selectivity and create novel aTFs responsive to HMA by directed evolution. We established a PobR mutant library with a capacity of 550,000 mutants using error-prone PCR and Megawhop PCR. Through our screening, two mutants were obtained with responsiveness to HMA. Analysis of each missense mutation indicating residues 122–126 were involved in its PobR ligand specificity. These results showed the effectiveness of directed evolution in switching the ligand specificity of a biosensor and improving HMA production. © 2022, The Author(s), under exclusive licence to Springer Nature B.V. | |
650 | 0 | 4 | |a 4-hydroxybenzoic acids |
650 | 0 | 4 | |a Acid production |
650 | 0 | 4 | |a Biosensors |
650 | 0 | 4 | |a Directed evolution |
650 | 0 | 4 | |a Directed evolution |
650 | 0 | 4 | |a Error prone PCR |
650 | 0 | 4 | |a Hydroxymandelic acid |
650 | 0 | 4 | |a Hydroxy-mandelic acid |
650 | 0 | 4 | |a Hydroxy-mandelic acid |
650 | 0 | 4 | |a Ligand specificity |
650 | 0 | 4 | |a Ligand specificity |
650 | 0 | 4 | |a Ligands |
650 | 0 | 4 | |a Mandelic acid |
650 | 0 | 4 | |a Mutant libraries |
650 | 0 | 4 | |a Organic acids |
650 | 0 | 4 | |a Pobr |
650 | 0 | 4 | |a PobR |
650 | 0 | 4 | |a Transcription |
650 | 0 | 4 | |a Transcription factors |
700 | 1 | 0 | |a Chen, F. |e author |
700 | 1 | 0 | |a Dong, Y. |e author |
700 | 1 | 0 | |a Fu, H. |e author |
700 | 1 | 0 | |a Guo, S. |e author |
700 | 1 | 0 | |a Liang, Y.Y. |e author |
700 | 1 | 0 | |a Luo, J. |e author |
700 | 1 | 0 | |a Su, K. |e author |
700 | 1 | 0 | |a Sui, G. |e author |
700 | 1 | 0 | |a Wang, P. |e author |
700 | 1 | 0 | |a Wang, Z. |e author |
700 | 1 | 0 | |a Yang, C. |e author |
700 | 1 | 0 | |a Zhang, B. |e author |
700 | 1 | 0 | |a Zhang, Y. |e author |
773 | |t World Journal of Microbiology and Biotechnology |