Rewiring the native methanol assimilation metabolism by incorporating the heterologous ribulose monophosphate cycle into Methylorubrum extorquens

Yuan, Xiao-Jie; Chen, Wenjing; Ma, Zhenhua; Yuan, Qianqian; Zhang, Min; He, Lian; Mo, Xuhua; Zhang, Chong; Zhang, Chang-Tai; Wang, Meng-Ying; Xing, Xin‐Hui; Yang, Song

doi:10.1016/j.ymben.2021.01.009

Cited by 27 publications

(20 citation statements)

References 69 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Expressing B. methanolicus derived hps and phi genes for completing the RuMP cycle and pfk for flux distribution from F6P, the engineered strain showed an increased cell growth and methanol consumption rate. Using this newly engineered chassis of M. extorquens AM1, 3-HP production was increased up to 0.857 g/L in the fed-batch bioreactor condition ( Yuan et al, 2021 ).…”

Section: Targeting Natural Methylotrophs For Chemical Productionmentioning

confidence: 99%

Developing methylotrophic microbial platforms for a methanol-based bioindustry

Singh

Kang

Kwon

et al. 2022

Front. Bioeng. Biotechnol.

View full text Add to dashboard Cite

Methanol, a relatively cheap and renewable single-carbon feedstock, has gained considerable attention as a substrate for the bio-production of commodity chemicals. Conventionally produced from syngas, along with emerging possibilities of generation from methane and CO2, this C1 substrate can serve as a pool for sequestering greenhouse gases while supporting a sustainable bio-economy. Methylotrophic organisms, with the inherent ability to use methanol as the sole carbon and energy source, are competent candidates as platform organisms. Accordingly, methanol bioconversion pathways have been an attractive target for biotechnological and bioengineering interventions in developing microbial cell factories. This review summarizes the recent advances in methanol-based production of various bulk and value-added chemicals exploiting the native and synthetic methylotrophic organisms. Finally, the current challenges and prospects of streamlining these methylotrophic platforms are discussed.

show abstract

Section: Targeting Natural Methylotrophs For Chemical Productionmentioning

confidence: 99%

Developing methylotrophic microbial platforms for a methanol-based bioindustry

Singh

Kang

Kwon

et al. 2022

Front. Bioeng. Biotechnol.

View full text Add to dashboard Cite

show abstract

“…Methanol can be produced from the greenhouse gases methane and CO 2 , thus conversion of methanol to valueadded chemicals via green biological processes may provide an attractive approach toward carbon neutrality (Zhu et al, 2020). Although natural methylotrophs such as Methylobacterium OPEN ACCESS EDITED BY Tian-Qiong Shi, Nanjing Normal University, China extorquens (Yuan et al, 2021) and Bacillus methanolicus (Brito et al, 2021) have been engineered to produce several chemicals, the relatively slow cell growth, incomprehensive understanding of cellular metabolism and the lack of effective genetic engineering tools significantly hinder the systematic engineering of these organisms toward real industrial applications (Zhu et al, 2020). Alternatively, engineering of well-characterized fast-growing microbial chassis such as Escherichia coli as synthetic methylotrophs by introducing heterologous methanol assimilation pathways has attracted broad attention in recent years (Gregory et al, 2022;Keller et al, 2022).…”

Section: Introductionmentioning

confidence: 99%

Engineering and adaptive laboratory evolution of Escherichia coli for improving methanol utilization based on a hybrid methanol assimilation pathway

Sun

Liu

Chen

2023

Front. Bioeng. Biotechnol.

View full text Add to dashboard Cite

Engineering Escherichia coli for efficient methanol assimilation is important for developing methanol as an emerging next-generation feedstock for industrial biotechnology. While recent attempts to engineer E. coli as a synthetic methylotroph have achieved great success, most of these works are based on the engineering of the prokaryotic ribulose monophosphate (RuMP) pathway. In this study, we introduced a hybrid methanol assimilation pathway which consists of prokaryotic methanol dehydrogenase (Mdh) and eukaryotic xylulose monophosphate (XuMP) pathway enzyme dihydroxyacetone synthase (Das) into E. coli and reprogrammed E. coli metabolism to improve methanol assimilation by combining rational design and adaptive laboratory evolution. By deletion and down-regulation of key genes in the TCA cycle and glycolysis to increase the flux toward the cyclic XuMP pathway, methanol consumption and the assimilation of methanol to biomass were significantly improved. Further improvements in methanol utilization and cell growth were achieved via adaptive laboratory evolution and a final evolved strain can grow on methanol with only 0.1 g/L yeast extract as co-substrate. 13C-methanol labeling assay demonstrated significantly higher labeling in intracellular metabolites in glycolysis, TCA cycle, pentose phosphate pathway, and amino acids. Transcriptomics analysis showed that the expression of fba, dhak, and part of pentose phosphate pathway genes were highly up-regulated, suggesting that the rational engineering strategies and adaptive evolution are effective for activating the cyclic XuMP pathway. This study demonstrated the feasibility and provided new strategies to construct synthetic methylotrophy of E. coli based on the hybrid methanol assimilation pathway with Mdh and Das.

show abstract

“…In contrast, methanol has been considered as a promising fermentation substrate due to its liquid properties and high reduction potential. − Moreover, methanol can be massively produced from CO 2 via the liquid sunlight route, which enables simultaneous carbon capture and solar energy storage. Recently, a methylotrophic bacteria Methylorubrum extorquens was harnessed for 3-HP production by engineering the methanol utilization pathways, which improved 3-HP production from 69 to 91 mg/L . This low 3-HP titer in methylotrophic bacteria was supposed to be attributed to inefficient methanol assimilation (<20 mmol/gDCW/h).…”

Section: Introductionmentioning

confidence: 99%

“…Recently, a methylotrophic bacteria Methylorubrum extorquens was harnessed for 3-HP production by engineering the methanol utilization pathways, which improved 3-HP production from 69 to 91 mg/L. 25 This low 3-HP titer in methylotrophic bacteria was supposed to be attributed to inefficient methanol assimilation (<20 mmol/gDCW/h). Alternatively, methylotrophic yeasts, such as Pichia pastoris, have high efficiency in methanol utilization (up to 100 mmol/ gDCW/h) and tolerance to harsh conditions, making them ideal hosts for methanol biotransformation.…”

Section: ■ Introductionmentioning

confidence: 99%

Efficient Bioproduction of 3-Hydroxypropionic Acid from Methanol by a Synthetic Yeast Cell Factory

Cai

Gao

et al. 2023

ACS Sustainable Chem. Eng.

View full text Add to dashboard Cite

Methanol is an ideal feedstock for bio-manufacturing chemicals without the dependence of sugars and the competition of arable lands. We here engineered an industrial yeast Pichia pastoris to efficiently produce 3-hydroxypropionic acid (3-HP) from sole methanol as a carbon source by using a malonyl-CoA-derived pathway. Optimizing the expression of malonyl-CoA reductase gene MCR from Chloroflexus aurantiacus and enhancing the supply of precursors and NADPH enabled 3-HP production of 1.5 g/L. To avoid the time-consuming genetic manipulation, metabolically transforming a free fatty acid (FFA)-overproducing strain toward 3-HP biosynthesis results in a 3-HP production of 1.9 g/L in a shake flask. Through further downregulation of methanol dissimilation, 3-HP production was improved to 2.2 g/L. Subsequent fed-batch cultivation in bioreactors achieved a remarkable 3-HP production of 48.2 g/L from minimal medium with a yield of 0.23 g/g methanol. Notably, this represents the highest reported 3-HP production from one-carbon (C1) feedstocks and is comparable to that from sugar in yeast. The high-level 3-HP production from methanol highlights the potential of P. pastoris as a workhorse for methanol biotransformation. Furthermore, the strategies presented in this study could be applied for production of other acetyl-CoA derivatives from methanol in P. pastoris.

show abstract

Rewiring the native methanol assimilation metabolism by incorporating the heterologous ribulose monophosphate cycle into Methylorubrum extorquens

Cited by 27 publications

References 69 publications

Developing methylotrophic microbial platforms for a methanol-based bioindustry

Developing methylotrophic microbial platforms for a methanol-based bioindustry

Engineering and adaptive laboratory evolution of Escherichia coli for improving methanol utilization based on a hybrid methanol assimilation pathway

Efficient Bioproduction of 3-Hydroxypropionic Acid from Methanol by a Synthetic Yeast Cell Factory

Contact Info

Product

Resources

About