Abstract
Biological ethylene production is a promising sustainable alternative approach for fossil-based ethylene production. The high glucose utilization of Z. mobilis makes it as a promising bioethylene producer. In this study, Zymomonas mobilis has been engineered to produce ethylene through the introduction of the synthetic ethylene-forming enzyme (EFE). We also investigated the effect of systematically knocking out the competitive metabolic pathway of pyruvate in an effort to improve the availability of pyruvate for ethylene production in Z. mobilis expressing EFE. Guided by these results, we tested a number of conjectures that could improve the α-ketoglutarate supply. Optimization of these pathways and different substrate supplies resulted in a greater production of ethylene (from 1.36 to 12.83 nmol/OD600/mL), which may guide future engineering work on ethylene production using other organisms. Meanwhile, we achieved an ethylene production of 5.8 nmol/OD600/mL in the ZM532-efe strain using enzymatic straw hydrolysate of corn straw as the sole carbon source. As a preferred host in biorefinery technologies using lignocellulosic biomass as feedstock, heterologous expression of EFE in Z. mobilis converts the non-ethylene producing strain into an ethylene-producing one using a metabolic engineering approach, which is of great significance for the utilization of cellulosic biomass in the future.
Key points
• Heterologous expression of EFE in Z. mobilis successfully converted the non-ethylene producing strain into an ethylene producer (1.36 nmol/OD600/mL). Targeted modifications of the central carbon metabolism can effectively improve ethylene production (peak production: 8.3 nmol/OD600/mL).
• The addition of nutrients to the medium can further increase the production of ethylene (peak production: 12.8 nmol/OD600/mL).
• The ZM532-efe strain achieved an ethylene production of 5.8 nmol/OD600/mL when enzymatic hydrolysate of corn straw was used as the sole carbon source.
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Data availability
The Z. mobilis 532 has been deposited at Guangdong Microbial Culture Center (GDMCC) under the Accession Number GDMCC60527.
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Funding
This research was financially supported by the National Natural Sciences Foundation of China (Grant No. 32070036), the Elite Program and Basic Research Program of Chinese Academy of Agricultural Sciences, and Central Public-interest Scientific Institution Basal Research Fund (No. Y2019XK23-01) to Mingxiong He.
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MXH designed the whole study and edited the entire draft manuscript. YH participated in all experiments and data collection. BW participated in data analysis. WX, KYZ, QY, TQ, QHY, and PTL participated in plasmid construction, RT-PCR and HPLC analysis. GQH participated in helpful discussions regarding the manuscript. All authors read and approved the final manuscript.
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He, Y., Wu, B., Xia, W. et al. Metabolic engineering of Zymomonas moblis for ethylene production from straw hydrolysate. Appl Microbiol Biotechnol 105, 1709–1720 (2021). https://doi.org/10.1007/s00253-021-11091-7
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DOI: https://doi.org/10.1007/s00253-021-11091-7