An in vitro synthetic biosystem based on acetate for production of phloroglucinol
Abstract Background Phloroglucinol is an important chemical, and the biosynthesis processes which can convert glucose to phloroglucinol have been established. However, due to approximate 80% of the glucose being transformed into undesirable by-products and biomass, this biosynthesis process only sho...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2017-08-01
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| Series: | BMC Biotechnology |
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| Online Access: | http://link.springer.com/article/10.1186/s12896-017-0376-z |
| _version_ | 1818340830120771584 |
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| author | Rubing Zhang Wei Liu Yujin Cao Xin Xu Mo Xian Huizhou Liu |
| author_facet | Rubing Zhang Wei Liu Yujin Cao Xin Xu Mo Xian Huizhou Liu |
| author_sort | Rubing Zhang |
| collection | DOAJ |
| description | Abstract Background Phloroglucinol is an important chemical, and the biosynthesis processes which can convert glucose to phloroglucinol have been established. However, due to approximate 80% of the glucose being transformed into undesirable by-products and biomass, this biosynthesis process only shows a low yield with the highest value of about 0.20 g/g. The industrial applications are usually hindered by the low current productivity and yield and also by the high costs. Generally, several different aspects limit the development of phloroglucinol biosynthesis. The yield of phloroglucinol is one of the most important parameters for its bioconversion especially from economic and ecological points of view. The in vitro biosynthesis of bio-based chemicals, is a flexible alternative with potentially high-yield to in vivo biosynthetic technology. Results By comparing the activity of acetyl-CoA synthetase (ACS) from Escherichia coli and Acetobacter pasteurianus, the highly active ACS2 was identified in A. pasteurianus. Acetyl-CoA carboxylase (ACC) from Acinetobacter calcoaceticus and phloroglucinol synthase (PhlD) from Pseudomonas fluorescens pf-5 were expressed and purified. Acetate was successfully transformed into phloroglucinol by the combined activity of above-mentioned enzymes and required cofactor. After optimization of the in vitro reaction system, phloroglucinol was then produced with a yield of nearly 0.64 g phloroglucinol/g acetic acid, which was equal to 91.43% of the theoretically possible maximum. Conclusions In this work, a novel in vitro synthetic system for a highly efficient production of phloroglucinol from acetate was demonstrated. The system’s performance suggests that in vitro synthesis of phloroglucinol has some advantages and is potential to become a feasible industrial alternative. Based on the results presented herewith, it is believed that in vitro biosystem will provide a feasible option for production of important industrial chemicals from acetate, which could work as a versatile biosynthetic platform. |
| first_indexed | 2024-12-13T15:49:08Z |
| format | Article |
| id | doaj.art-8dfa5a34614c406886619d4d6de6ba03 |
| institution | Directory Open Access Journal |
| issn | 1472-6750 |
| language | English |
| last_indexed | 2024-12-13T15:49:08Z |
| publishDate | 2017-08-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Biotechnology |
| spelling | doaj.art-8dfa5a34614c406886619d4d6de6ba032022-12-21T23:39:35ZengBMCBMC Biotechnology1472-67502017-08-011711810.1186/s12896-017-0376-zAn in vitro synthetic biosystem based on acetate for production of phloroglucinolRubing Zhang0Wei Liu1Yujin Cao2Xin Xu3Mo Xian4Huizhou Liu5CAS Key Laboratory of Bio-Based Materials, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of SciencesCAS Key Laboratory of Bio-Based Materials, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of SciencesCAS Key Laboratory of Bio-Based Materials, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of SciencesCAS Key Laboratory of Bio-Based Materials, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of SciencesCAS Key Laboratory of Bio-Based Materials, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of SciencesCAS Key Laboratory of Bio-Based Materials, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of SciencesAbstract Background Phloroglucinol is an important chemical, and the biosynthesis processes which can convert glucose to phloroglucinol have been established. However, due to approximate 80% of the glucose being transformed into undesirable by-products and biomass, this biosynthesis process only shows a low yield with the highest value of about 0.20 g/g. The industrial applications are usually hindered by the low current productivity and yield and also by the high costs. Generally, several different aspects limit the development of phloroglucinol biosynthesis. The yield of phloroglucinol is one of the most important parameters for its bioconversion especially from economic and ecological points of view. The in vitro biosynthesis of bio-based chemicals, is a flexible alternative with potentially high-yield to in vivo biosynthetic technology. Results By comparing the activity of acetyl-CoA synthetase (ACS) from Escherichia coli and Acetobacter pasteurianus, the highly active ACS2 was identified in A. pasteurianus. Acetyl-CoA carboxylase (ACC) from Acinetobacter calcoaceticus and phloroglucinol synthase (PhlD) from Pseudomonas fluorescens pf-5 were expressed and purified. Acetate was successfully transformed into phloroglucinol by the combined activity of above-mentioned enzymes and required cofactor. After optimization of the in vitro reaction system, phloroglucinol was then produced with a yield of nearly 0.64 g phloroglucinol/g acetic acid, which was equal to 91.43% of the theoretically possible maximum. Conclusions In this work, a novel in vitro synthetic system for a highly efficient production of phloroglucinol from acetate was demonstrated. The system’s performance suggests that in vitro synthesis of phloroglucinol has some advantages and is potential to become a feasible industrial alternative. Based on the results presented herewith, it is believed that in vitro biosystem will provide a feasible option for production of important industrial chemicals from acetate, which could work as a versatile biosynthetic platform.http://link.springer.com/article/10.1186/s12896-017-0376-zIn vitroAcetateAcetyl-CoA synthetasePhloroglucinolHigh yield |
| spellingShingle | Rubing Zhang Wei Liu Yujin Cao Xin Xu Mo Xian Huizhou Liu An in vitro synthetic biosystem based on acetate for production of phloroglucinol BMC Biotechnology In vitro Acetate Acetyl-CoA synthetase Phloroglucinol High yield |
| title | An in vitro synthetic biosystem based on acetate for production of phloroglucinol |
| title_full | An in vitro synthetic biosystem based on acetate for production of phloroglucinol |
| title_fullStr | An in vitro synthetic biosystem based on acetate for production of phloroglucinol |
| title_full_unstemmed | An in vitro synthetic biosystem based on acetate for production of phloroglucinol |
| title_short | An in vitro synthetic biosystem based on acetate for production of phloroglucinol |
| title_sort | in vitro synthetic biosystem based on acetate for production of phloroglucinol |
| topic | In vitro Acetate Acetyl-CoA synthetase Phloroglucinol High yield |
| url | http://link.springer.com/article/10.1186/s12896-017-0376-z |
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