Cargando…
Directed evolution of the fusion enzyme for improving astaxanthin biosynthesis in Saccharomyces cerevisiae
The accumulation of the intermediate zeaxanthin and canthaxanthin in the astaxanthin biosynthesis pathway catalyzed by β-carotene hydroxylase (crtZ) and β-carotene ketolase (crtW) decreases the content of the astaxanthin. Here, we exploited directed evolution of the fusion of crtZ and crtW for impro...
Autores principales: | , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
KeAi Publishing
2022
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9668533/ https://www.ncbi.nlm.nih.gov/pubmed/36408203 http://dx.doi.org/10.1016/j.synbio.2022.10.005 |
_version_ | 1784831934366482432 |
---|---|
author | Ding, Yong-Wen Lu, Chuan-Zhen Zheng, Yan Ma, Han-Zhang Jin, Jin Jia, Bin Yuan, Ying-Jin |
author_facet | Ding, Yong-Wen Lu, Chuan-Zhen Zheng, Yan Ma, Han-Zhang Jin, Jin Jia, Bin Yuan, Ying-Jin |
author_sort | Ding, Yong-Wen |
collection | PubMed |
description | The accumulation of the intermediate zeaxanthin and canthaxanthin in the astaxanthin biosynthesis pathway catalyzed by β-carotene hydroxylase (crtZ) and β-carotene ketolase (crtW) decreases the content of the astaxanthin. Here, we exploited directed evolution of the fusion of crtZ and crtW for improving astaxanthin biosynthesis in Saccharomyces cerevisiae. The results demonstrated that the fusion enzyme of crtZ-crtW with 2 X GGGGS peptides linker can effectively reduce the accumulation of intermediates and improves the content of astaxanthin. Compared with the control strain, the fusion enzyme of ketase and hydroxylase reduced zeaxanthin and canthaxanthin by 7 and 14 times and increased astaxanthin by 1.6 times, respectively. Moreover, 9 variant fusion mutants with improved astaxanthin production were generated through directed evolution. Combining these dominant mutants generated a variant, L95S + I206L, which increased the astaxanthin content of 3.8 times than the control strain. The AlphaFold2 assisted structural analysis indicated that these two mutations alter the interaction between the substrate and the enzymes pocket. Our research provided an efficient idea to reduce the accumulation of the intermediate products in complex biosynthesis pathway. |
format | Online Article Text |
id | pubmed-9668533 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | KeAi Publishing |
record_format | MEDLINE/PubMed |
spelling | pubmed-96685332022-11-18 Directed evolution of the fusion enzyme for improving astaxanthin biosynthesis in Saccharomyces cerevisiae Ding, Yong-Wen Lu, Chuan-Zhen Zheng, Yan Ma, Han-Zhang Jin, Jin Jia, Bin Yuan, Ying-Jin Synth Syst Biotechnol Original Research Article The accumulation of the intermediate zeaxanthin and canthaxanthin in the astaxanthin biosynthesis pathway catalyzed by β-carotene hydroxylase (crtZ) and β-carotene ketolase (crtW) decreases the content of the astaxanthin. Here, we exploited directed evolution of the fusion of crtZ and crtW for improving astaxanthin biosynthesis in Saccharomyces cerevisiae. The results demonstrated that the fusion enzyme of crtZ-crtW with 2 X GGGGS peptides linker can effectively reduce the accumulation of intermediates and improves the content of astaxanthin. Compared with the control strain, the fusion enzyme of ketase and hydroxylase reduced zeaxanthin and canthaxanthin by 7 and 14 times and increased astaxanthin by 1.6 times, respectively. Moreover, 9 variant fusion mutants with improved astaxanthin production were generated through directed evolution. Combining these dominant mutants generated a variant, L95S + I206L, which increased the astaxanthin content of 3.8 times than the control strain. The AlphaFold2 assisted structural analysis indicated that these two mutations alter the interaction between the substrate and the enzymes pocket. Our research provided an efficient idea to reduce the accumulation of the intermediate products in complex biosynthesis pathway. KeAi Publishing 2022-11-10 /pmc/articles/PMC9668533/ /pubmed/36408203 http://dx.doi.org/10.1016/j.synbio.2022.10.005 Text en © 2022 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Original Research Article Ding, Yong-Wen Lu, Chuan-Zhen Zheng, Yan Ma, Han-Zhang Jin, Jin Jia, Bin Yuan, Ying-Jin Directed evolution of the fusion enzyme for improving astaxanthin biosynthesis in Saccharomyces cerevisiae |
title | Directed evolution of the fusion enzyme for improving astaxanthin biosynthesis in Saccharomyces cerevisiae |
title_full | Directed evolution of the fusion enzyme for improving astaxanthin biosynthesis in Saccharomyces cerevisiae |
title_fullStr | Directed evolution of the fusion enzyme for improving astaxanthin biosynthesis in Saccharomyces cerevisiae |
title_full_unstemmed | Directed evolution of the fusion enzyme for improving astaxanthin biosynthesis in Saccharomyces cerevisiae |
title_short | Directed evolution of the fusion enzyme for improving astaxanthin biosynthesis in Saccharomyces cerevisiae |
title_sort | directed evolution of the fusion enzyme for improving astaxanthin biosynthesis in saccharomyces cerevisiae |
topic | Original Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9668533/ https://www.ncbi.nlm.nih.gov/pubmed/36408203 http://dx.doi.org/10.1016/j.synbio.2022.10.005 |
work_keys_str_mv | AT dingyongwen directedevolutionofthefusionenzymeforimprovingastaxanthinbiosynthesisinsaccharomycescerevisiae AT luchuanzhen directedevolutionofthefusionenzymeforimprovingastaxanthinbiosynthesisinsaccharomycescerevisiae AT zhengyan directedevolutionofthefusionenzymeforimprovingastaxanthinbiosynthesisinsaccharomycescerevisiae AT mahanzhang directedevolutionofthefusionenzymeforimprovingastaxanthinbiosynthesisinsaccharomycescerevisiae AT jinjin directedevolutionofthefusionenzymeforimprovingastaxanthinbiosynthesisinsaccharomycescerevisiae AT jiabin directedevolutionofthefusionenzymeforimprovingastaxanthinbiosynthesisinsaccharomycescerevisiae AT yuanyingjin directedevolutionofthefusionenzymeforimprovingastaxanthinbiosynthesisinsaccharomycescerevisiae |