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Combinatorial expression of different β-carotene hydroxylases and ketolases in Escherichia coli for increased astaxanthin production

  • Yuanqing Wu
  • Panpan Yan
  • Xuewei Liu
  • Zhiwen Wang
  • Ya-Jie Tang
  • Tao ChenEmail author
  • Xueming Zhao
Metabolic Engineering and Synthetic Biology - Original Paper
  • 33 Downloads

Abstract

In natural produced bacteria, β-carotene hydroxylase (CrtZ) and β-carotene ketolase (CrtW) convert β-carotene into astaxanthin. To increase astaxanthin production in heterologous strain, simple and effective strategies based on the co-expression of CrtZ and CrtW were applied in E. coli. First, nine artificial operons containing crtZ and crtW genes from different sources were constructed and, respectively, introduced into E. coli ZF237T, a β-carotene producing host. Among the nine resulting strains, five accumulated detectable amounts of astaxanthin ranging from 0.49 to 8.07 mg/L. Subsequently, the protein fusion CrtZ to CrtW using optimized peptide linkers further increased the astaxanthin production. Strains expressing fusion proteins with CrtZ rather than CrtW attached to the N-terminus accumulated much more astaxanthin. The astaxanthin production of the best strain ZF237T/CrtZAs-(GS)1-WBs was 127.6% and 40.2% higher than that of strains ZF237T/crtZAsWBs and ZF237T/crtZBsWPs, respectively. The strategies depicted here also will be useful for the heterologous production of other natural products.

Keywords

Astaxanthin Combinatorial expression Artificial operon Fusion protein Bifunctional enzymes 

Notes

Funding

This work was supported by the National Natural Science Foundation of China (NSFC-21621004, NSFC-21776208 and NSFC-21776209).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants performed by any of the authors.

Supplementary material

10295_2019_2214_MOESM1_ESM.docx (1.1 mb)
Supplementary material 1 (DOCX 1159 kb)

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Copyright information

© Society for Industrial Microbiology and Biotechnology 2019

Authors and Affiliations

  1. 1.Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), School of Chemical Engineering and TechnologyTianjin UniversityTianjinPeople’s Republic of China
  2. 2.State Key Laboratory of Microbial TechnologyShandong UniversityQingdaoPeople’s Republic of China
  3. 3.Key Laboratory of Fermentation Engineering (Ministry of Education), Hubei Key Laboratory of Industrial Microbiology, Hubei Provincial Cooperative Innovation Center of Industrial FermentationHubei University of TechnologyWuhanPeople’s Republic of China

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