Three steps in one pot: biosynthesis of 4-hydroxycinnamyl alcohols using immobilized whole cells of two genetically engineered Escherichia coli strains
Abstract Background 4-Hydroxycinnamyl alcohols are a class of natural plant secondary metabolites that include p-coumaryl alcohol, caffeyl alcohol, coniferyl alcohol and sinapyl alcohol, and have physiological, ecological and biomedical significance. While it is necessary to investigate the biologic...
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BMC
2017-06-01
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Series: | Microbial Cell Factories |
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Online Access: | http://link.springer.com/article/10.1186/s12934-017-0722-9 |
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author | Shuxin Liu Jiabin Liu Jiayin Hou Nan Chao Ying Gai Xiangning Jiang |
author_facet | Shuxin Liu Jiabin Liu Jiayin Hou Nan Chao Ying Gai Xiangning Jiang |
author_sort | Shuxin Liu |
collection | DOAJ |
description | Abstract Background 4-Hydroxycinnamyl alcohols are a class of natural plant secondary metabolites that include p-coumaryl alcohol, caffeyl alcohol, coniferyl alcohol and sinapyl alcohol, and have physiological, ecological and biomedical significance. While it is necessary to investigate the biological pathways and economic value of these alcohols, research is hindered because of their limited availability and high cost. Traditionally, these alcohols are obtained by chemical synthesis and plant extraction. However, synthesis by biotransformation with immobilized microorganisms is of great interest because it is environmentally friendly and offers high stability and regenerable cofactors. Therefore, we produced 4-hydroxycinnamyl alcohols using immobilized whole cells of engineered Escherichia coli as the biocatalyst. Results In this study, we used the recombinant E. coli strain, M15–4CL1–CCR, expressing the fusion protein 4-coumaric acid: coenzyme A ligase and the cinnamoyl coenzyme A reductase and a recombinant E. coli strain, M15–CAD, expressing cinnamyl alcohol dehydrogenase from Populus tomentosa (P. tomentosa). High performance liquid chromatography and mass spectrometry showed that the immobilized whole cells of the two recombinant E. coli strains could effectively convert the phenylpropanoic acids to their corresponding 4-hydroxycinnamyl alcohols. Further, the optimum buffer pH and the reaction temperature were pH 7.0 and 30 °C. Under these conditions, the molar yield of the p-coumaryl alcohol, the caffeyl alcohol and the coniferyl alcohol was around 58, 24 and 60%, respectively. Moreover, the highly sensitive and selective HPLC–PDA–ESI–MSn method used in this study could be applied to the identification and quantification of these aromatic polymers. Conclusions We have developed a dual-cell immobilization system for the production of 4-hydroxycinnamyl alcohols from inexpensive phenylpropanoic acids. This biotransformation method is both simple and environmental-friendly, which is promising for the practical and cost effective synthesis of natural products. Graphical abstract Biotransformation process of phenylpropanoic acids by immobilized whole-cells |
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issn | 1475-2859 |
language | English |
last_indexed | 2024-04-12T13:06:56Z |
publishDate | 2017-06-01 |
publisher | BMC |
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series | Microbial Cell Factories |
spelling | doaj.art-4ebab84112f3449e9453da536d4f51e82022-12-22T03:32:00ZengBMCMicrobial Cell Factories1475-28592017-06-0116111210.1186/s12934-017-0722-9Three steps in one pot: biosynthesis of 4-hydroxycinnamyl alcohols using immobilized whole cells of two genetically engineered Escherichia coli strainsShuxin Liu0Jiabin Liu1Jiayin Hou2Nan Chao3Ying Gai4Xiangning Jiang5College of Biological Science and Technology, Beijing Forestry UniversityCollege of Biological Science and Technology, Beijing Forestry UniversityCollege of Biological Science and Technology, Beijing Forestry UniversityCollege of Biological Science and Technology, Beijing Forestry UniversityCollege of Biological Science and Technology, Beijing Forestry UniversityCollege of Biological Science and Technology, Beijing Forestry UniversityAbstract Background 4-Hydroxycinnamyl alcohols are a class of natural plant secondary metabolites that include p-coumaryl alcohol, caffeyl alcohol, coniferyl alcohol and sinapyl alcohol, and have physiological, ecological and biomedical significance. While it is necessary to investigate the biological pathways and economic value of these alcohols, research is hindered because of their limited availability and high cost. Traditionally, these alcohols are obtained by chemical synthesis and plant extraction. However, synthesis by biotransformation with immobilized microorganisms is of great interest because it is environmentally friendly and offers high stability and regenerable cofactors. Therefore, we produced 4-hydroxycinnamyl alcohols using immobilized whole cells of engineered Escherichia coli as the biocatalyst. Results In this study, we used the recombinant E. coli strain, M15–4CL1–CCR, expressing the fusion protein 4-coumaric acid: coenzyme A ligase and the cinnamoyl coenzyme A reductase and a recombinant E. coli strain, M15–CAD, expressing cinnamyl alcohol dehydrogenase from Populus tomentosa (P. tomentosa). High performance liquid chromatography and mass spectrometry showed that the immobilized whole cells of the two recombinant E. coli strains could effectively convert the phenylpropanoic acids to their corresponding 4-hydroxycinnamyl alcohols. Further, the optimum buffer pH and the reaction temperature were pH 7.0 and 30 °C. Under these conditions, the molar yield of the p-coumaryl alcohol, the caffeyl alcohol and the coniferyl alcohol was around 58, 24 and 60%, respectively. Moreover, the highly sensitive and selective HPLC–PDA–ESI–MSn method used in this study could be applied to the identification and quantification of these aromatic polymers. Conclusions We have developed a dual-cell immobilization system for the production of 4-hydroxycinnamyl alcohols from inexpensive phenylpropanoic acids. This biotransformation method is both simple and environmental-friendly, which is promising for the practical and cost effective synthesis of natural products. Graphical abstract Biotransformation process of phenylpropanoic acids by immobilized whole-cellshttp://link.springer.com/article/10.1186/s12934-017-0722-94-Hydroxycinnamyl alcohols4-Coumaric acid: coenzyme A ligaseCinnamoyl coenzyme A reductaseCinnamyl alcohol dehydrogenaseImmobilized whole-cellEscherichia coli |
spellingShingle | Shuxin Liu Jiabin Liu Jiayin Hou Nan Chao Ying Gai Xiangning Jiang Three steps in one pot: biosynthesis of 4-hydroxycinnamyl alcohols using immobilized whole cells of two genetically engineered Escherichia coli strains Microbial Cell Factories 4-Hydroxycinnamyl alcohols 4-Coumaric acid: coenzyme A ligase Cinnamoyl coenzyme A reductase Cinnamyl alcohol dehydrogenase Immobilized whole-cell Escherichia coli |
title | Three steps in one pot: biosynthesis of 4-hydroxycinnamyl alcohols using immobilized whole cells of two genetically engineered Escherichia coli strains |
title_full | Three steps in one pot: biosynthesis of 4-hydroxycinnamyl alcohols using immobilized whole cells of two genetically engineered Escherichia coli strains |
title_fullStr | Three steps in one pot: biosynthesis of 4-hydroxycinnamyl alcohols using immobilized whole cells of two genetically engineered Escherichia coli strains |
title_full_unstemmed | Three steps in one pot: biosynthesis of 4-hydroxycinnamyl alcohols using immobilized whole cells of two genetically engineered Escherichia coli strains |
title_short | Three steps in one pot: biosynthesis of 4-hydroxycinnamyl alcohols using immobilized whole cells of two genetically engineered Escherichia coli strains |
title_sort | three steps in one pot biosynthesis of 4 hydroxycinnamyl alcohols using immobilized whole cells of two genetically engineered escherichia coli strains |
topic | 4-Hydroxycinnamyl alcohols 4-Coumaric acid: coenzyme A ligase Cinnamoyl coenzyme A reductase Cinnamyl alcohol dehydrogenase Immobilized whole-cell Escherichia coli |
url | http://link.springer.com/article/10.1186/s12934-017-0722-9 |
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