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Overview of piceatannol biosynthesis pathway genes from p-coumaric acid. The biosynthesis pathway genes and precursors biosynthesis genes are divided into three modules. Module I contains 4-coumarate: CoA ligase and stilbene synthase genes, module II contains various genes for biosynthesis of acetyl-CoA and malonyl-CoA, and module III contains resveratrol 3′-hydroxylase to convert resveratrol to piceatannol. The bold arrow reactions catalyzing genes were overexpressed in different combinations. Enzymes catalyzing each step are given in blue color. Each module is highlighted in different colored boxes
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Resveratrol and its ortho-hydroxylated derivative piceatannol were biosynthesized by modular pathway engineering in Escherichia coli. The biosynthetic pathway was divided into three different modules. Module I includes polyketide biosynthetic genes; module II genes include acetyl-CoA and malonyl-CoA pool-enhancing genes from three different organis...
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Citations
... The resulting C. glutamicum strain accumulated 112 mg L −1 of resveratrol from glucose, without supplementation of phenylpropanoid precursor molecules or any inhibitors of fatty acid synthesis. Using a modular pathway engineering strategy, Shrestha et al. (2018) also increased the resveratrol titer in E. coli (137 mg L −1 ) by over-expressing the malonate assimilation pathway genes matB and matC from Streptomyces coelicolor A3(2) and acetyl-CoA complex genes (accCADB) from E. coli BL21 and Nocardia farcinica IFM10152, to enhance the pool of intracellular malonyl-CoA and acetyl-CoA, respectively. ...
Aromatic compounds are an important class of chemicals with different industrial applications. They are usually produced by chemical synthesis from petroleum-derived feedstocks, such as toluene, xylene and benzene. However, we are now facing threats from the excessive use of fossil fuels causing environmental problems such as global warming. Furthermore, fossil resources are not infinite, and will ultimately be depleted. To cope with these problems, the sustainable production of aromatic chemicals from renewable non-food biomass is urgent. With this in mind, the search for alternative methodologies to produce aromatic compounds using low-cost and environmentally friendly processes is becoming more and more important. Microorganisms are able to produce aromatic and aromatic-derivative compounds from sugar-based carbon sources. Metabolic engineering strategies as well as bioprocess optimization enable the development of microbial cell factories capable of efficiently producing aromatic compounds. This review presents current breakthroughs in microbial production of specialty aromatic and aromatic-derivative products, providing an overview on the general strategies and methodologies applied to build microbial cell factories for the production of these compounds. We present and describe some of the current challenges and gaps that must be overcome in order to render the biotechnological production of specialty aromatic and aromatic-derivative attractive and economically feasible at industrial scale.
... Another alternative is the use of genetically modified plants (Hain and Grimmig 2000;Giovinazzo et al. 2005) and microorganisms (Beekwilder et al. 2006;C. G. Lim et al. 2011;Shrestha et al. 2018;Kallscheuer et al. 2016) transformed via genetic engineering with the genes implied in the stilbene synthesis route. Chemically, stilbenes can be synthesized by Heck, Perkin or Witting reactions, among others (Tian and Liu 2020). ...
Stilbenes are phenolic compounds naturally synthesized as secondary metabolites by the shikimate pathway in plants. Research on them has increased in recent years due to their therapeutic potential as antioxidant, antimicrobial, anti-inflammatory, anticancer, cardioprotective and anti-obesity agents. Amongst them, resveratrol has attracted the most attention, although there are other natural and synthesized stilbenes with enhanced properties. However, stilbenes have some physicochemical and pharmacokinetic problems that need to be overcome before considering their applications. Human clinical evidence of their bioactivity is still controversial due to this fact and hence, exhaustive basis science on stilbenes is needed before applied science. This review gathers the main physicochemical and biological properties of natural stilbenes, establishes structure-activity relationships among them, emphasizing the current problems that limit their applications and presenting some promising approaches to overcome these issues: the encapsulation in different agents and the structural modification to obtain novel stilbenes with better features. The bioactivity of stilbenes should move from promising to evident.
... Under these conditions, a higher yield of 86.1% was obtained within 3 h, translating to a productivity of 1.38 g L −1 h −1 . This result is much higher than all those reported in literature obtained using different biocatalysts (for comparison, see previous refs [30][31][32][33] and the table 1 in our previous paper, 11 except the one recently obtained in our laboratory with TCHCs as the catalyst (1.92 g −1 L −1 h −1 ). 11 So far, more than a dozen studies have successfully demonstrated the production of Pic from Res by means of biotechnological strategies, most of which are based on the use of metabolic engineering [30][31][32][33] and whole-cell catalysis via heterogeneous expression in Escherichia coli of tyrosinase or other monooxygenases. ...
... This result is much higher than all those reported in literature obtained using different biocatalysts (for comparison, see previous refs [30][31][32][33] and the table 1 in our previous paper, 11 except the one recently obtained in our laboratory with TCHCs as the catalyst (1.92 g −1 L −1 h −1 ). 11 So far, more than a dozen studies have successfully demonstrated the production of Pic from Res by means of biotechnological strategies, most of which are based on the use of metabolic engineering [30][31][32][33] and whole-cell catalysis via heterogeneous expression in Escherichia coli of tyrosinase or other monooxygenases. [34][35][36] Both whole-cell catalysts and free enzymes have two critical drawbacks: the catalyst is not immobilized and hence cannot be recycled, and the productivity is usually low because of the poor conversion yield as a consequence of the long operational period required. ...
BACKGROUND
Metal–organic frameworks (MOFs) have gained increasing attention with ever‐expanding applications. Developing new MOF‐based immobilized enzymes with new applications is required, not only for demonstrating the generality and applicability of using MOFs for enzyme immobilization, but also to provide potential biocatalysts for industrial applications. To the best of the authors’ knowledge, this is the first report of immobilizing mushroom tyrosinase on zeolitic imidazolate frameworks (ZIFs), with two new applications being developed.
RESULTS
Upon immobilization through a one‐pot in situ encapsulation approach, the resultant tyrosinase@ZIF‐8 was characterized in structural features and catalytic properties. It was much more stable against pH and temperature relative to the free enzyme. The new biocatalyst was highly efficient in catalyzing the synthesis of catecholic products with pharmacological benefits (l‐DOPA, piceatannol and 3′‐hydroxypterostilbene) and in eliminating phenolic pollutants (phenol, p‐cresol, p‐chlorophenol). Excellent productivities were obtained for the synthesis of the three catecholic products (0.14, 1.38 and 1.46 g L⁻¹ h⁻¹, respectively). A complete removal of the three phenolic pollutants was achieved within 2.5 h, superior to other processes mediated by the same enzyme, or others, immobilized on different supports. The reusability of the new biocatalyst can be remarkably improved by entrapment into polyvinyl alcohol‐alginate gel.
CONCLUSIONS
Tyrosinase can be immobilized on a new type of MOF with advantages such as easy preparation and excellent catalytic performance. This novel immobilization strategy for tyrosinase offers an excellent biocatalyst potent for both catecholic product synthesis and phenolic wastewater treatment. © 2021 Society of Chemical Industry (SCI).
... Piceatannol was synthesized from glucose in E. coli with a yield of 21.5 mg/L [16]. Overexpression of acetate and malonate assimilation pathways along with additional supply of malonate in the culture medium yielded 124 mg/L of piceatannol (3-hydroxyresveratrol) in E. coli [17]. ...
Resveratrol is a typical plant phenolic compound whose derivatives are synthesized through hydroxylation, O -methylation, prenylation, and oligomerization. Resveratrol and its derivatives exhibit anti-neurodegenerative, anti-rheumatoid, and anti-inflammatory effects. Owing to the diverse biological activities of these compounds and their importance in human health, this study attempted to synthesize five resveratrol derivatives (isorhapontigenin, pterostilbene, 4-methoxyresveratrol, piceatannol, and rhapontigenin) using Escherichia coli . Two-culture system was used to improve the final yield of resveratrol derivatives. Resveratrol was synthesized in the first E. coli cell that harbored genes for resveratrol biosynthesis including TAL (tyrosine ammonia lyase), 4CL (4-coumaroyl CoA ligase), STS (stilbene synthase) and genes for tyrosine biosynthesis such as aroG (deoxyphosphoheptonate aldolase) and tyrA (prephenate dehydrogenase). Thereafter, culture filtrate from the first cell was used for the modification reaction carried out using the second E. coli harboring hydroxylase and/or O -methyltransferase. Approximately, 89.8 mg/L of resveratrol was synthesized and using the same, five derivatives were prepared with a conversion rate of 88.2% to 22.9%. Using these synthesized resveratrol derivatives, we evaluated their anti-inflammatory activity. 4-Methoxyresveratrol, pterostilbene and isorhapontigenin showed the anti-inflammatory effects without any toxicity. In addition, pterostilbene exhibited the enhanced anti-inflammatory effects for macrophages compared to resveratrol.
... Furthermore, the synthesis of tryptophan is regulated by various regulatory mechanisms, such as feedback repression, feedback inhibition and feed-forward inhibition [10,11] which increase the difficulty of tryptophan production. With the development of biotechnology and the proposal of new strategies, including modular metabolic engineering [12,13], discovery of new mutation points for removing feedback inhibition [14] and cofactor engineering [15], it is possible to further enhance the L-tryptophan production by Escherichia coli. ...
Acetic acid, an important by-product of L-tryptophan production by Escherichia coli, is detrimental to cell growth and tryptophan accumulation. To reduce acetic acid accumulation and further improve the production of L-tryptophan, an acetyl-coenzyme A (acetyl-CoA) synthase (ACS) with high synthetic activity is needed. In this study, E. coli ACS was found to have a superior catalytic capability compared to that of other bacterial strains. To reduce the accumulation of acetic acid by metabolic overflow, a higher expression level of ACS was obtained by promoter engineering and copy number optimisation. Furthermore, reconstruction of the tri-carboxylic acid cycle (TCA) not only enhanced the supply of phosphoenolpyruvate (PEP), which is one of the precursors required for tryptophan synthesis, but also improved bacterial growth. The final engineered strain T16 accumulated 54.6 g/L of L-tryptophan, 11.4% more than the original strain T03, with an accumulation of only 1.4 g/L acetic acid after fermentation, which reduced by 74.5% compared to T03.
Abbreviations
acetyl-CoA
acetyl-coenzyme A
ACS
acetyl-coenzyme A synthase
TCA
tricarboxylic acid cycle
PEP
phosphoenolpyruvate
HMP
hexose monophosphate pathway;
ackA
acetate kinase
pta
phosphotransacetylase
AMP
adenosine monophosphate
ATP
adenosine triphosphate
... There are other strategies as well that focus on rerouting the endogenous malonyl-CoA pathway flow for the stoichiometric modeling, resulting in a higher resveratrol production by increasing the malonyl-CoA pool [133]. Besides these, there are other strategies as aforementioned, such as inhibiting the fab operon using antisense RNA [14,129] and downregulating the expression of fatty-acid biosynthesis by using the CRISPRi system [65], and overexpressing of ACC, acs (acetate assimilation enzyme) [134] along with the deletion of pta, ackA, and adhE [96] which helps to increase the cytosolic malonyl-CoA pool. Moreover, malonyl-CoA is also increased by overexpressing the matB and matC genes encoding malonate-CoA synthase and malonate carrier proteins, respectively [134,135]. ...
... Besides these, there are other strategies as aforementioned, such as inhibiting the fab operon using antisense RNA [14,129] and downregulating the expression of fatty-acid biosynthesis by using the CRISPRi system [65], and overexpressing of ACC, acs (acetate assimilation enzyme) [134] along with the deletion of pta, ackA, and adhE [96] which helps to increase the cytosolic malonyl-CoA pool. Moreover, malonyl-CoA is also increased by overexpressing the matB and matC genes encoding malonate-CoA synthase and malonate carrier proteins, respectively [134,135]. Recently, a modular engineering approach was used to increase the pool of malonyl-CoA by overexpressing acetate as well as malonate assimilation pathway genes from three different sources. Along with 4CL, STS, and HpaBC enzymes encoding genes, significant amounts of resveratrol and its hydroxylated derivative piceatannol were produced [134]. ...
... Recently, a modular engineering approach was used to increase the pool of malonyl-CoA by overexpressing acetate as well as malonate assimilation pathway genes from three different sources. Along with 4CL, STS, and HpaBC enzymes encoding genes, significant amounts of resveratrol and its hydroxylated derivative piceatannol were produced [134]. ...
The very well-known bioactive natural product, resveratrol (3,5,4′-trihydroxystilbene), is a highly studied secondary metabolite produced by several plants, particularly grapes, passion fruit, white tea, and berries. It is in high demand not only because of its wide range of biological activities against various kinds of cardiovascular and nerve-related diseases, but also as important ingredients in pharmaceuticals and nutritional supplements. Due to its very low content in plants, multi-step isolation and purification processes, and environmental and chemical hazards issues, resveratrol extraction from plants is difficult, time consuming, impracticable, and unsustainable. Therefore, microbial hosts, such as Escherichia coli, Saccharomyces cerevisiae, and Corynebacterium glutamicum, are commonly used as an alternative production source by improvising resveratrol biosynthetic genes in them. The biosynthesis genes are rewired applying combinatorial biosynthetic systems, including metabolic engineering and synthetic biology, while optimizing the various production processes. The native biosynthesis of resveratrol is not present in microbes, which are easy to manipulate genetically, so the use of microbial hosts is increasing these days. This review will mainly focus on the recent biotechnological advances for the production of resveratrol, including the various strategies used to produce its chemically diverse derivatives.
... contains supplementary material, which is available to authorized users. friendliness (Furuya et al. 2018;Shrestha et al. 2018). In particular, biocatalysts that hydroxylate resveratrol to piceatannol are useful in piceatannol production (Fig. 1). ...
Piceatannol is a valuable natural polyphenol with therapeutic potential in cardiovascular and metabolic disease treatment. In this study, we screened for microorganisms capable of producing piceatannol from resveratrol via regioselective hydroxylation. In the first screening, we isolated microorganisms utilizing resveratrol, phenol, or 4-hydroxyphenylacetic acid as a carbon source for growth. In the second screening, we assayed the isolated microorganisms for hydroxylation of resveratrol. Using this screening procedure, a variety of resveratrol-converting microorganisms were obtained. One Gram-negative bacterium, Ensifer sp. KSH1, and one Gram-positive bacterium, Arthrobacter sp. KSH3, utilized 4-hydroxyphenylacetic acid as a carbon source for growth and efficiently hydroxylated resveratrol to piceatannol without producing any detectable by-products. The hydroxylation activity of strains KSH1 and KSH3 was strongly induced by cultivation with 4-hydroxyphenylacetic acid as a carbon source during stationary growth phase. Using the 4-hydroxyphenylacetic acid–induced cells as a biocatalyst under optimal conditions, production of piceatannol by strains KSH1 and KSH3 reached 3.6 mM (0.88 g/L) and 2.6 mM (0.64 g/L), respectively. We also cloned genes homologous to the monooxygenase gene hpaBC from strains KSH1 and KSH3. Introduction of either hpaBC homolog into Escherichia coli endowed the host with resveratrol-hydroxylating activity.
... At the same time, the production of fine chemicals including natural products by E. coli is rapidly developing. Naringenin (Whitaker et al., 2017), astaxanthin , glucoraphanin (Yang et al., 2017a), arbutin (Shen et al., 2017), β-carotene , lycopene (Wu et al., 2018), resveratrol (Shrestha et al., 2018), isoprene , and other natural products (Greunke et al., 2018) can now be produced by engineered E. coli. However, often this is accompanied by low titers. ...
Biotechnological production of fuels and chemicals from renewable resources is an appealing way to move from the current petroleum-based economy to a biomass-based green economy. Recently, the feedstocks that can be used for bioconversion or fermentation have been expanded to plant biomass, microbial biomass, and industrial waste. Several microbes have been engineered to produce chemicals from renewable resources, among which Escherichia coli is one of the best studied. Much effort has been made to engineer E. coli to produce fuels and chemicals from different renewable resources. In this paper, we focused on E. coli and systematically reviewed a range of fuels and chemicals that can be produced from renewable resources by engineered E. coli. Moreover, we proposed how can we further improve the efficiency for utilizing renewable resources by engineered E. coli, and how can we engineer E. coli for utilizing alternative renewable feedstocks. e.g. C1 gases and methanol. This review will help the readers better understand the current progress in this field and provide insights for further metabolic engineering efforts in E. coli.
... The authors also stated that Sam5 enzyme from Saccharothrix espanaensis exhibited 5.7-fold higher conversion rate of resveratrol to piceatannol, compared to coumarate 3-hydroxylase. Similarly, Shrestha et al. (2018) devised the modular pathway engineering in E. coli for the production of piceatannol. The biosynthetic pathway genes 4-CL from Parsley, STS from V. vinifera, hpaBC from E. coli, matB, and matC from Streptomyces coelicolor were assembled in different fashion in modular approach and used p-coumaric acid as basic substrate. ...
Resveratrol (3,5,4′-trihydroxystilbene) and piceatannol (3,5,3′,4′-tetrahydroxystilbene) are well-known natural products that are produced by plants. They are important ingredients in pharmaceutical industries and nutritional supplements. They display a wide spectrum of biological activity. Thus, the needs for these compounds are increasing. The natural products have been found in diverse plants, mostly such as grapes, passion fruit, white tea, berries, and many more. The extraction of these products from plants is quite impractical because of the low production in plants, downstream processing difficulties, chemical hazards, and environmental issues. Thus, alternative production in microbial hosts has been devised with combinatorial biosynthetic systems, including metabolic engineering, synthetic biology, and optimization in production process. Since the biosynthesis is not native in microbial hosts such as Escherichia coli, Saccharomyces cerevisiae, and Corynebacterium glutamicum, genetic engineering and manipulation have made it possible. In this review, the discussion will mainly focus on recent progress in production of resveratrol and piceatannol, including the various strategies used for their production.
... Recombinant bacteria (E. coli) or yeast (Yarrowia lipolytica) indeed displayed an increase of their intracellular malonyl-CoA pool upon expression of the malonyl-CoA biosynthetic operon including matB and matC genes (Huang et al., 2009;Shrestha et al., 2018;Wu et al., 2013 and (Figs. 2 and 3). ...
... Using this successful strategy, the pinosylvin titer reached 281 mg/L (Table 2). Very recently, a similar approach was applied for the production of resveratrol and piceatannol in E. coli (Shrestha et al., 2018). The employed strategy targeted the intracellular pool of acetyl-CoA and malonyl-CoA stressing on the malonate assimilation and anabolic pathways by overexpression of matB and matC genes or the ACC gene. ...
... The combination of modules 1 and 2c upon supplementation with disodium malonate led to a resveratrol titer of 151 mg/L. High piceatannol yield (124 mg/L) was obtained from resveratrol after the addition of module 3 to the latter construct (Table 2) (Shrestha et al., 2018). ...
Numerous in vitro and in vivo studies on biological activities of phytostilbenes have brought to the fore the remarkable properties of these compounds and their derivatives, making them a top storyline in natural product research fields. However, getting stilbenes in sufficient amounts for routine biological activity studies and make them available for the pharmaceutical and/or nutraceutical industry applications, is hampered by the difficulty to source them through synthetic chemistry-based pathways or extraction from the native plants. Hence, microbial cell cultures have rapidly became potent workhorse factories for stilbene production. In this review, we present the combined efforts made during the past 15 years to engineer stilbene metabolic pathways in microbial cells, mainly the Saccharomyces cerevisiae baker yeast, the Escherichia coli and the Corynebacterium glutamicum bacteria. Rationalized approaches to the heterologous expression of the partial or the entire stilbene biosynthetic routes will be described, allowing the identification and bypassing of the major bottlenecks in the endogenous microbial cell metabolism as well as the tight cellular regulations of the genes involved in these metabolic pathways. The contributions of bioinformatics to synthetic biology are developed to highlight their tremendous help in predicting which target genes are likely to be up-regulated or deleted for controlling the dynamics of precursor flows in the tailored microbial cells. Further insight is given to the metabolic engineering of microbial cells with “decorating” enzymes, such as methyl and glycosyltransferases or hydroxylases, which can act sequentially on the stilbene core structure. Altogether, the cellular optimization of stilbene biosynthetic pathways integrating more and more complex constructs up to twelve genetic modifications has led to stilbene titers ranging from hundreds of milligrams to the gram-scale yields from various carbon sources. Through this review, the microbial production of stilbenes is analyzed, stressing both the engineering dynamic regulation of biosynthetic pathways and the endogenous control of stilbene precursors.
