scholarly journals De novo biosynthesis of bioactive isoflavonoids by engineered yeast cell factories

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Quanli Liu ◽  
Yi Liu ◽  
Gang Li ◽  
Otto Savolainen ◽  
Yun Chen ◽  
...  
Keyword(s):  
Natural Products ◽  
Yeast Cell ◽  
De Novo ◽  
Structural Complexity ◽  
Value Added ◽  
Fine Tuning ◽  
Attractive Alternative ◽  
Efficient Production ◽  
De Novo Biosynthesis ◽  
Cell Factories

AbstractIsoflavonoids comprise a class of plant natural products with great nutraceutical, pharmaceutical and agricultural significance. Their low abundance in nature and structural complexity however hampers access to these phytochemicals through traditional crop-based manufacturing or chemical synthesis. Microbial bioproduction therefore represents an attractive alternative. Here, we engineer the metabolism of Saccharomyces cerevisiae to become a platform for efficient production of daidzein, a core chemical scaffold for isoflavonoid biosynthesis, and demonstrate its application towards producing bioactive glucosides from glucose, following the screening-reconstruction-application engineering framework. First, we rebuild daidzein biosynthesis in yeast and its production is then improved by 94-fold through screening biosynthetic enzymes, identifying rate-limiting steps, implementing dynamic control, engineering substrate trafficking and fine-tuning competing metabolic processes. The optimized strain produces up to 85.4 mg L−1 of daidzein and introducing plant glycosyltransferases in this strain results in production of bioactive puerarin (72.8 mg L−1) and daidzin (73.2 mg L−1). Our work provides a promising step towards developing synthetic yeast cell factories for de novo biosynthesis of value-added isoflavonoids and the multi-phased framework may be extended to engineer pathways of complex natural products in other microbial hosts.

Insights on the Advancements of In Silico Metabolic Studies of Succinic Acid Producing Microorganisms: A Review with Emphasis on Actinobacillus succinogenes

Fermentation ◽  
2021 ◽  
Vol 7 (4) ◽  
pp. 220
Author(s):  
Wubliker Dessie ◽  
Zongcheng Wang ◽  
Xiaofang Luo ◽  
Meifeng Wang ◽  
Zuodong Qin
Keyword(s):  
Succinic Acid ◽  
In Silico ◽  
Value Added ◽  
Research Progress ◽  
Efficient Production ◽  
Actinobacillus Succinogenes ◽  
Cell Factories ◽  
In Silico Studies ◽  
Current Scenario ◽  
Critical Problems

Succinic acid (SA) is one of the top candidate value-added chemicals that can be produced from biomass via microbial fermentation. A considerable number of cell factories have been proposed in the past two decades as native as well as non-native SA producers. Actinobacillus succinogenes is among the best and earliest known natural SA producers. However, its industrial application has not yet been realized due to various underlying challenges. Previous studies revealed that the optimization of environmental conditions alone could not entirely resolve these critical problems. On the other hand, microbial in silico metabolic modeling approaches have lately been the center of attention and have been applied for the efficient production of valuable commodities including SA. Then again, literature survey results indicated the absence of up-to-date reviews assessing this issue, specifically concerning SA production. Hence, this review was designed to discuss accomplishments and future perspectives of in silico studies on the metabolic capabilities of SA producers. Herein, research progress on SA and A. succinogenes, pathways involved in SA production, metabolic models of SA-producing microorganisms, and status, limitations and prospects on in silico studies of A. succinogenes were elaborated. All in all, this review is believed to provide insights to understand the current scenario and to develop efficient mathematical models for designing robust SA-producing microbial strains.

Metabolic engineering of glycosylated polyketide biosynthesis

2018 ◽  
Vol 2 (3) ◽  
pp. 389-403 ◽  
Author(s):  
Ramesh Prasad Pandey ◽  
Prakash Parajuli ◽  
Jae Kyung Sohng
Keyword(s):  
Natural Products ◽  
Metabolic Engineering ◽  
Value Added ◽  
Chemical Diversity ◽  
Yeast Cells ◽  
Microbial Cell Factories ◽  
Cell Factories ◽  
Modification Method ◽  
Post Modification ◽  
Microbial Biosynthesis

Microbial cell factories are extensively used for the biosynthesis of value-added chemicals, biopharmaceuticals, and biofuels. Microbial biosynthesis is also realistic for the production of heterologous molecules including complex natural products of plant and microbial origin. Glycosylation is a well-known post-modification method to engineer sugar-functionalized natural products. It is of particular interest to chemical biologists to increase chemical diversity of molecules. Employing the state-of-the-art systems and synthetic biology tools, a range of small to complex glycosylated natural products have been produced from microbes using a simple and sustainable fermentation approach. In this context, this review covers recent notable metabolic engineering approaches used for the biosynthesis of glycosylated plant and microbial polyketides in different microorganisms. This review article is broadly divided into two major parts. The first part is focused on the biosynthesis of glycosylated plant polyketides in prokaryotes and yeast cells, while the second part is focused on the generation of glycosylated microbial polyketides in actinomycetes.

scholarly journals Divergent synthesis of complex diterpenes through a hybrid oxidative approach

Science ◽  
2020 ◽  
Vol 369 (6505) ◽  
pp. 799-806 ◽  
Author(s):  
Xiao Zhang ◽  
Emma King-Smith ◽  
Liao-Bin Dong ◽  
Li-Cheng Yang ◽  
Jeffrey D. Rudolf ◽  
...  
Keyword(s):  
Natural Products ◽  
De Novo ◽  
Biological Activities ◽  
Structural Complexity ◽  
Enzymatic Oxidation ◽  
Chemical Tools ◽  
Skeletal Rearrangements ◽  
Skeletal Diversity

Polycyclic diterpenes exhibit many important biological activities, but de novo synthetic access to these molecules is highly challenging because of their structural complexity. Semisynthetic access has also been limited by the lack of chemical tools for scaffold modifications. We report a chemoenzymatic platform to access highly oxidized diterpenes by a hybrid oxidative approach that strategically combines chemical and enzymatic oxidation methods. This approach allows for selective oxidations of previously inaccessible sites on the parent carbocycles and enables abiotic skeletal rearrangements to additional underlying architectures. We synthesized a total of nine complex natural products with rich oxygenation patterns and skeletal diversity in 10 steps or less from ent-steviol.

scholarly journals The Role of Synthetic Biology in the Design of Microbial Cell Factories for Biofuel Production

2011 ◽  
Vol 2011 ◽  
pp. 1-9 ◽  
Author(s):  
Verónica Leticia Colin ◽  
Analía Rodríguez ◽  
Héctor Antonio Cristóbal
Keyword(s):  
Synthetic Biology ◽  
Industrial Applications ◽  
Microbial Cell ◽  
Biodiesel Production ◽  
Biofuel Production ◽  
Attractive Alternative ◽  
Efficient Production ◽  
Microbial Cell Factories ◽  
Cell Factories

Insecurity in the supply of fossil fuels, volatile fuel prices, and major concerns regarding climate change have sparked renewed interest in the production of fuels from renewable resources. Because of this, the use of biodiesel has grown dramatically during the last few years and is expected to increase even further in the future. Biodiesel production through the use of microbial systems has marked a turning point in the field of biofuels since it is emerging as an attractive alternative to conventional technology. Recent progress in synthetic biology has accelerated the ability to analyze, construct, and/or redesign microbial metabolic pathways with unprecedented precision, in order to permit biofuel production that is amenable to industrial applications. The review presented here focuses specifically on the role of synthetic biology in the design of microbial cell factories for efficient production of biodiesel.

scholarly journals Cell-based and cell-free biocatalysis for the production of d-glucaric acid

2020 ◽  
Vol 13 (1) ◽  
Author(s):  
Lu-Zhou Chen ◽  
Si-Ling Huang ◽  
Jin Hou ◽  
Xue-Ping Guo ◽  
Feng-Shan Wang ◽  
...  
Keyword(s):  
Metabolic Pathways ◽  
De Novo ◽  
Value Added ◽  
Host Cells ◽  
Efficient Production ◽  
Glucaric Acid ◽  
A Value ◽  
Potential Applications ◽  
Increasing Demand

Abstractd-Glucaric acid (GA) is a value-added chemical produced from biomass, and has potential applications as a versatile platform chemical, food additive, metal sequestering agent, and therapeutic agent. Marketed GA is currently produced chemically, but increasing demand is driving the search for eco-friendlier and more efficient production approaches. Cell-based production of GA represents an alternative strategy for GA production. A series of synthetic pathways for GA have been ported into Escherichia coli, Saccharomyces cerevisiae and Pichia pastoris, respectively, and these engineered cells show the ability to synthesize GA de novo. Optimization of the GA metabolic pathways in host cells has leapt forward, and the titer and yield have increased rapidly. Meanwhile, cell-free multi-enzyme catalysis, in which the desired pathway is constructed in vitro from enzymes and cofactors involved in GA biosynthesis, has also realized efficient GA bioconversion. This review presents an overview of studies of the development of cell-based GA production, followed by a brief discussion of potential applications of biosensors that respond to GA in these biosynthesis routes.

scholarly journals Metabolic engineering Escherichia coli for efficient production of icariside D2

2019 ◽  
Vol 12 (1) ◽  
Author(s):  
Xue Liu ◽  
Lingling Li ◽  
Jincong Liu ◽  
Jianjun Qiao ◽  
Guang-Rong Zhao
Keyword(s):  
Escherichia Coli ◽  
De Novo ◽  
Carbon Sources ◽  
Gene Expression Pattern ◽  
Fine Tuning ◽  
Cell Factory ◽  
Efficient Production ◽  
Uridine Diphosphate ◽  
Microbial Cell Factory ◽  
E Coli

Abstract Background Icariside D2 is a plant-derived natural glycoside with pharmacological activities of inhibiting angiotensin-converting enzyme and killing leukemia cancer cells. Production of icariside D2 by plant extraction and chemical synthesis is inefficient and environmentally unfriendly. Microbial cell factory offers an attractive route for economical production of icariside D2 from renewable and sustainable bioresources. Results We metabolically constructed the biosynthetic pathway of icariside D2 in engineered Escherichia coli. We screened the uridine diphosphate glycosyltransferases (UGTs) and obtained an active RrUGT3 that regio-specifically glycosylated tyrosol at phenolic position to exclusively synthesize icariside D2. We put heterologous genes in E. coli cell for the de novo biosynthesis of icariside D2. By fine-tuning promoter and copy number as well as balancing gene expression pattern to decrease metabolic burden, the BMD10 monoculture was constructed. Parallelly, for balancing pathway strength, we established the BMT23–BMD12 coculture by distributing the icariside D2 biosynthetic genes to two E. coli strains BMT23 and BMD12, responsible for biosynthesis of tyrosol from preferential xylose and icariside D2 from glucose, respectively. Under the optimal conditions in fed-batch shake-flask fermentation, the BMD10 monoculture produced 3.80 g/L of icariside D2 using glucose as sole carbon source, and the BMT23–BMD12 coculture produced 2.92 g/L of icariside D2 using glucose–xylose mixture. Conclusions We for the first time reported the engineered E. coli for the de novo efficient production of icariside D2 with gram titer. It would be potent and sustainable approach for microbial production of icariside D2 from renewable carbon sources. E. coli–E. coli coculture approach is not limited to glycoside production, but could also be applied to other bioproducts.

scholarly journals Phenolic Amides Are Potent Inhibitors ofDe NovoNucleotide Biosynthesis

2015 ◽  
Vol 81 (17) ◽  
pp. 5761-5772 ◽  
Author(s):  
Tippapha Pisithkul ◽  
Tyler B. Jacobson ◽  
Thomas J. O'Brien ◽  
David M. Stevenson ◽  
Daniel Amador-Noguez
Keyword(s):  
Phenolic Compounds ◽  
De Novo ◽  
Plant Biomass ◽  
Value Added ◽  
Metabolic Effects ◽  
Efficient Production ◽  
Biosynthetic Pathways ◽  
Nucleotide Biosynthesis ◽  
Phenolic Amides ◽  
The Impact

ABSTRACTAn outstanding challenge toward efficient production of biofuels and value-added chemicals from plant biomass is the impact that lignocellulose-derived inhibitors have on microbial fermentations. Elucidating the mechanisms that underlie their toxicity is critical for developing strategies to overcome them. Here, usingEscherichia colias a model system, we investigated the metabolic effects and toxicity mechanisms of feruloyl amide and coumaroyl amide, the predominant phenolic compounds in ammonia-pretreated biomass hydrolysates. Using metabolomics, isotope tracers, and biochemical assays, we showed that these two phenolic amides act as potent and fast-acting inhibitors of purine and pyrimidine biosynthetic pathways. Feruloyl or coumaroyl amide exposure leads to (i) a rapid buildup of 5-phosphoribosyl-1-pyrophosphate (PRPP), a key precursor in nucleotide biosynthesis, (ii) a rapid decrease in the levels of pyrimidine biosynthetic intermediates, and (iii) a long-term generalized decrease in nucleotide and deoxynucleotide levels. Tracer experiments using13C-labeled sugars and [15N]ammonia demonstrated that carbon and nitrogen fluxes into nucleotides and deoxynucleotides are inhibited by these phenolic amides. We found that these effects are mediated via direct inhibition of glutamine amidotransferases that participate in nucleotide biosynthetic pathways. In particular, feruloyl amide is a competitive inhibitor of glutamine PRPP amidotransferase (PurF), which catalyzes the first committed step inde novopurine biosynthesis. Finally, external nucleoside supplementation prevents phenolic amide-mediated growth inhibition by allowing nucleotide biosynthesis via salvage pathways. The results presented here will help in the development of strategies to overcome toxicity of phenolic compounds and facilitate engineering of more efficient microbial producers of biofuels and chemicals.

scholarly journals De Novo Biosynthesis of C-Arabinosylated Flavones by Utilization of Indica Rice C-Glycosyltransferases

2021 ◽  
Author(s):  
Zhuo Chen ◽  
Yuwei Sun ◽  
Guangyi Wang ◽  
Ying Zhang ◽  
Qian Zhang ◽  
...  
Keyword(s):  
De Novo ◽  
Indica Rice ◽  
Fed Batch ◽  
Rice Varieties ◽  
E Coli ◽  
De Novo Biosynthesis ◽  
Microbial Cell Factories ◽  
Cell Factories ◽  
Fed Batch Fermentation ◽  
Heterologous Pathway

Abstract Flavone C-arabinosides/xylosides are plant-originated glycoconjugates with various bioactivities. However, the potential utility of these molecules is hindered by their low abundance in nature. Engineering biosynthesis pathway in heterologous bacterial chassis provides a sustainable source of these C-glycosides. We previously reported bifunctional C-glucosyl/C-arabinosyltransferases in Oryza sativa japonica and O. sativa indica, which influence the C-glycoside spectrum in different rice varieties. In this study, we proved the C-arabinosyltransferring activity of rice C-glycosyltransferases (CGTs) on the mono-C-glucoside substrate nothofagin, followed by taking advantage of specific CGTs and introducing heterologous UDP-pentose supply, to realize the production of eight different C-arabinosides/xylosides in recombinant E. coli. Fed-batch fermentation and precursor supplement maximized the titer of rice-originated C-arabinosides to 20~110 mg/L in an E. coli chassis. The optimized final titer of schaftoside and apigenin di-C-arabinoside reached 19.87 and 113.16 mg/L respectively. We demonstrate here the success of de novo bio-production of C-arabinosylated and C-xylosylated flavones by heterologous pathway reconstitution. These results lay a foundation for further optimal manufacture of complex flavonoid compounds in microbial cell factories.

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