Biosensor-assisted transcriptional regulator engineering for Methylobacterium extorquens AM1 to improve mevalonate synthesis by increasing the acetyl-CoA supply

Liang, Wenzhong; Cui, Lanyu; Cui, Jinyu; Yu, Kaiwen; Yang, Song; Wang, Tianmin; Guan, Changge; Zhang, Chong; Xing, Xin‐Hui

doi:10.1016/j.ymben.2016.11.010

Cited by 53 publications

(42 citation statements)

References 45 publications

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“…Taken together, the generated essentiality data extend our understanding of methylotrophy. At the same time, the data provide a basis for metabolic engineering to convert methanol into value-added products in M. extorquens itself [50,[52][53][54] and beyond for transplanting methylotrophy to established platform organisms [6,7].…”

Section: Discussionmentioning

confidence: 91%

“…Besides confirming the link between Prk and QscR, the proteome data also allowed to enlarge the known QscR regulon by additional genes, i.e., formate-tetrahydrofolate ligase ftfL and oxalyl-CoA reductase panE2 (Mext_2139) [49], including its neighboring genes (Mext_2138/40; Data S4). Due to the central regulatory function of QscR, the regulator is also of interest for re-engineering of M. extorquens, as was suggested recently [50]. Thus, tuning RuBP levels via Prk activity may provide an alternative engineering path for the production of value-added products.…”

Section: A Regulatory Role Of Phosphoribulokinase For One-carbon Assimentioning

confidence: 96%

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Transposon Sequencing Uncovers an Essential Regulatory Function of Phosphoribulokinase for Methylotrophy

Ochsner¹,

Christen²,

Hemmerle³

et al. 2017

Current Biology

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Methylotrophy is the ability of organisms to grow at the expense of reduced one-carbon compounds, such as methanol or methane. Here, we used transposon sequencing combining hyper-saturated transposon mutagenesis with high-throughput sequencing to define the essential methylotrophy genome of Methylobacterium extorquens PA1, a model methylotroph. To distinguish genomic regions required for growth only on methanol from general required genes, we contrasted growth on methanol with growth on succinate, a non-methylotrophic reference substrate. About 500,000 insertions were mapped for each condition, resulting in a median insertion distance of five base pairs. We identified 147 genes and 76 genes as specific for growth on methanol and succinate, respectively, and a set of 590 genes as required under both growth conditions. For the integration of metabolic functions, we reconstructed a genome-scale metabolic model and performed in silico essentiality analysis. In total, the approach uncovered 95 genes not previously described as crucial for methylotrophy, including genes involved in respiration, carbon metabolism, transport, and regulation. Strikingly, regardless of the absence of the Calvin cycle in the methylotroph, the screen led to the identification of the gene for phosphoribulokinase as essential during growth on methanol, but not during growth on succinate. Genetic experiments in addition to metabolomics and proteomics revealed that phosphoribulokinase serves a key regulatory function. Our data support a model according to which ribulose-1,5-bisphosphate is an essential metabolite that induces a transcriptional regulator driving one-carbon assimilation.

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Section: Discussionmentioning

confidence: 91%

Section: A Regulatory Role Of Phosphoribulokinase For One-carbon Assimentioning

confidence: 96%

Transposon Sequencing Uncovers an Essential Regulatory Function of Phosphoribulokinase for Methylotrophy

Ochsner¹,

Christen²,

Hemmerle³

et al. 2017

Current Biology

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“…5b) 76 . Through further optimization of the methanol utilization and isoprenoid pathways, production of mevalonate reached 2.7 g l -1 with a yield of 0.08 g per g methanol 77 . The methylotrophic yeast Pichia pastoris has also been engineered for production of lycopene 78 and nootkatone 79 .…”

Section: Engineering Utilization Of Single-carbon Feedstocksmentioning

confidence: 99%

Barriers and opportunities in bio-based production of hydrocarbons

2018

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Global climate change caused by the accumulation of greenhouse gases (GHGs) has caused concerns regarding the continued reliance on fossil fuels as our primary energy source. Hydrocarbons produced from biomass using microbial fermentation processes can serve as high-quality liquid transportation fuels and may contribute to a reduction in GHG emissions. Here, we discuss the barriers and opportunities for bio-based production of hydrocarbons to be used as diesel and jet fuels and review recent advances in engineering microbes for production of these chemicals. There are two main challenges associated with establishing bio-based hydrocarbon production from cheap feedstocks; lowering the cost of developing efficient and robust microbial cell factories and establishing more efficient routes for biomass hydrolysis to sugars for fermentation. We discuss how to develop novel systems and synthetic biology tools that can enable faster and cheaper construction of microbial cell factories and thereby address the first challenge, as well as recent advances in biomass processing that will likely lead to overcoming the second challenge in the near future.

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“…This method has enabled rapid screening and isolation of flavonoid-producing microbes without the use of conventional chromatography and enzymatic assays techniques that are costly, laborious and time-consuming Siedler et al 2014). The advantages of using biosensor-aided strain development have led to growing interest in implementing this strategy in metabolite analysis, product monitoring and metabolic engineering of natural products (Chou & Keasling 2013;Liang et al 2017;Rogers et al 2015).…”

Section: Ecolimentioning

confidence: 99%

Current Progress in Production of Flavonoids using Systems and Synthetic Biology Platforms

Bahaudin

Bazli

Baharum

et al. 2018

JSM

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Flavonoid is an industrially-important compound due to its high pharmaceutical and cosmeceutical values. However, conventional methods in extracting and synthesizing flavonoids are costly, laborious and not sustainable due to small amount of natural flavonoids, large amounts of chemicals and space used. Biotechnological production of flavonoids represents a viable and sustainable route especially through the use of metabolic engineering strategies in microbial production hosts. In this review, we will highlight recent strategies for the improving the production of flavonoids using synthetic biology approaches in particular the innovative strategies of genetically-encoded biosensors for in vivo metabolite analysis and high-throughput screening methods using fluorescence-activated cell sorting (FACS). Implementation of transcription factor based-biosensor for microbial flavonoid production and integration of systems and synthetic biology approaches for natural product development will also be discussed.

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Biosensor-assisted transcriptional regulator engineering for Methylobacterium extorquens AM1 to improve mevalonate synthesis by increasing the acetyl-CoA supply

Cited by 53 publications

References 45 publications

Transposon Sequencing Uncovers an Essential Regulatory Function of Phosphoribulokinase for Methylotrophy

Transposon Sequencing Uncovers an Essential Regulatory Function of Phosphoribulokinase for Methylotrophy

Barriers and opportunities in bio-based production of hydrocarbons

Current Progress in Production of Flavonoids using Systems and Synthetic Biology Platforms

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