Renewable biosynthesis of isoprene from wastewater through a synthetic biology approach: the role of individual organic compounds

doi:10.1007/s11783-024-1788-3

Front. Environ. Sci. Eng.

2024, Vol. 18

Issue (3) : 28 https://doi.org/10.1007/s11783-024-1788-3

RESEARCH ARTICLE

Renewable biosynthesis of isoprene from wastewater through a synthetic biology approach: the role of individual organic compounds

Min Yang¹, Xianghui Li¹, Weixiang Chao¹, Xiang Gao², Huan Wang³, Lu Lu¹(

)

¹. State Key Laboratory of Urban Water Resource and Environment, School of Civil and Environmental Engineering, Harbin Institute of Technology, Shenzhen 518055, China
². Center for Materials Synthetic Biology, CAS Key Laboratory of Quantitative Engineering Biology of CAS, Shenzhen Institute of Synthetic Biology, Shenzhen Institutes of Advanced Technology, Chinese Academic of Science, Shenzhen 518000, China
³. Institute of Urban Ecology and Environmental Technology, School of Materials and Environmental Engineering, Shenzhen Polytechnic University, Shenzhen 518055, China

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Abstract

● Engineered E. coli can use wastewater as the only feedstock to product isoprene.

● Glucose, maltose, glycerol and lactate can be used for isoprene biosynthesis.

● Starch, protein and acetate can’t feed the E. coli growth.

● The optimum C/N ratio and essential nutrients addition enhance isoprene yield.

● The cost and CO₂ emission are significantly reduced by using wastewater.

The biosynthesis of isoprene offers a more sustainable alternative to fossil fuel-based approaches, yet its success has been largely limited to pure organic compounds and the cost remains a challenge. This study proposes a waste-to-wealth strategy for isoprene biosynthesis utilizing genetically engineered E. coli bacteria to convert organic waste from real food wastewater. The impact of organic compounds present in wastewater on E. coli growth and isoprene production was systematically investigated. The results demonstrated that with filtration pretreatment of wastewater, isoprene yield, and production achieved 115 mg/g COD and 7.1 mg/(L·h), respectively. Moreover, even without pretreatment, isoprene yield only decreased by ~ 24%, indicating promising scalability. Glucose, maltose, glycerol, and lactate are effective substrates for isoprene biosynthesis, whereas starch, protein, and acetate do not support E. coli growth. The optimum C/N ratio for isoprene production was found to be 8:1. Furthermore, augmenting essential nutrients in wastewater elevated the isoprene yield increased to 159 mg/g COD. The wastewater biosynthesis significantly reduced the cost (44%–53% decrease, p-value < 0.01) and CO₂ emission (46%–55% decrease, p-value < 0.01) compared with both sugar fermentation and fossil fuel–based refining. This study introduced a more sustainable and economically viable approach to isoprene synthesis, offering an avenue for resource recovery from wastewater.

Keywords Wastewater Resource recovery Genetic engineering Biosynthesis Isoprene

Corresponding Author(s): Lu Lu

About author:

Peng Lei and Charity Ngina Mwangi contributed equally to this work.

Issue Date: 26 October 2023

Cite this article:

Min Yang,Xianghui Li,Weixiang Chao, et al. Renewable biosynthesis of isoprene from wastewater through a synthetic biology approach: the role of individual organic compounds[J]. Front. Environ. Sci. Eng., 2024, 18(3): 28.

URL:

https://academic.hep.com.cn/fese/EN/10.1007/s11783-024-1788-3
https://academic.hep.com.cn/fese/EN/Y2024/V18/I3/28

Fig.1 Schematic of isoprene biosynthesis using organics in wastewater as the only substrates.

Fig.2 Isoprene production by engineered E. coli using real organic wastewater. (a) Evaluation of cell growth based on OD₆₀₀ values, (b) cumulative isoprene production and yield, (c) chemical oxygen demand, and (d) carbon balance during 24 h of bacterial culture.

Fig.3 Isoprene production by engineered E. coli using individual organic substrates as carbon source. (a) Optical density at 600 nm (OD₆₀₀) measurements, (b) isoprene yield after 24 h of bacterial culture.

Fig.4 Isoprene production by engineered E. coli using artificial wastewater. (a) Evaluation of cell growth based on OD₆₀₀ values, (b) the cumulative isoprene production and yield, (c) carbon balance during bacterial culture, and (d) changes in concentration of each organic component in artificial wastewater over culture time.

Fig.5 Isoprene production by engineered E. coli under various C/N ratios. (a) Optical density at 600 nm (OD₆₀₀), (b) isoprene production using artificial wastewater with varying C/N ratios after 24 h of bacterial culture, and (c) isoprene production using real wastewater after adding various elements with concentrations equivalent to those in M9 medium after 24 h of bacterial culture.

Fig.6 Isoprene and CO₂ production after 48 h culture using wastewater with and without filtration pretreatment.

Fig.7 Economic and carbon emission evaluation. (a) Comparison of current pathways to isoprene production: fossil-fuels refining, bacteria fermentation of sugars, and engineered bacteria fermentation of wastewater, (b) system boundary for engineered bacteria fermentation of wastewater, and its comparison in terms of (c) economic cost and (d) carbon emissions with the current pathways.

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