One-pot, three-step cascade synthesis of D-danshensu using engineered Escherichia coli whole cells

Xiong, Tianzhen; Jia, Ping; Jiang, Jing; Bai, Yajun; Fan, Tai-Ping; Zheng, Xiaohui; Cai, Yujie

doi:10.1016/j.jbiotec.2019.05.008

Cited by 13 publications

(17 citation statements)

References 26 publications

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“…For these reasons, a cascade for the transformation of catechol and pyruvate to SAA via L-DOPA was designed. 11 The only limitation is that the reaction from catechol and pyruvate to L-DOPA is reversible, making the system require excess substrates. Here, for the biosynthesis of SAA, we selected a much less expensive starting compound, PPA, of the L-tyrosine/L-phenylalanine biosynthetic pathway, a powerful platform to produce aromatic compounds from renewable feedstocks.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

“…SAA is from the herbal plant Salvia miltiorrhiza and has potential pharmaceutical activities including antitumor activity, antioxidant activity, and anti-inflammatory properties, as well as improving cardiovascular diseases. 11 It also serves as a precursor for the synthesis of pharmaceutical compounds such as Danshensu Bingpian Zhi and isopropyl 3-(3,4-dihydroxyphenyl)-2-hydroxypropanoate. 12 Except for direct extraction from S. miltiorrhiza and chemical synthesis, engineered Escherichia coli has been used to synthesize SAA from 4-hydroxyphenylpyruvate (4HPP).…”

Section: ■ Introductionmentioning

confidence: 99%

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A Thermophilic Biofunctional Multienzyme Cascade Reaction for Cell-Free Synthesis of Salvianic Acid A and 3,4-Dihydroxymandelic Acid

Zhang

Jian

2019

ACS Sustainable Chem. Eng.

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The production of high-value phenolic acids is of significant importance for sustainability in the pharmaceutical industry. In this study, an artificial thermophilic reaction cascade composed of D-mandelate dehydrogenase (ManDH), phenylalanine 4-hydroxylase (PAH), and hydroxyphenylacetate 3-hydroxylase (HpaH) was constructed to utilize the lowcost phenylpyruvic acid (PPA) and 2-phenylglyoxylic acid (PGA) as dual substrates. The dihydroxyphenolic acids salvianic acid A (SAA) and 3,4-dihydroxymandelic acid (DOMA) were then produced at 84.9% and 90.9% of the theoretical yields from PPA and PGA, respectively. The key to success of the reaction was the novel TbManDH from Thermococcus barophilus and TcPAH from Thermomonospora curvata that exhibited a broad substrate specificity. The newly in situ regeneration of the cofactor 6,7-dimethyl-5,6,7,8-tetrahydropterine (DMPH 4 ) in combination with nicotinamide adenine dinucleotide (NADH) recycling also contributed to the high production yields. Considering the broad substrate specificity of many other natural enzymes, it is attractive to design such dual-or even multifunctional catalyst systems for efficiently producing value-added chemicals.

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Section: ■ Results and Discussionmentioning

confidence: 99%

Section: ■ Introductionmentioning

confidence: 99%

A Thermophilic Biofunctional Multienzyme Cascade Reaction for Cell-Free Synthesis of Salvianic Acid A and 3,4-Dihydroxymandelic Acid

Zhang

Jian

2019

ACS Sustainable Chem. Eng.

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“…The implementation of an in vivo multi-enzyme cascade system in a designer cell catalyst requires the fine tuning of expression levels. The precise co-expression strategy affects the expression of target enzymes and the catalytic efficiency of the co-expression system [17,21,31]. As shown in Scheme 1, in our study, the designed in vivo cascade route was mainly composed of two modules: an oxidative deamination module catalyzed by l-amino acid deaminase and a reductive amination module catalyzed by d-amino acid dehydrogenase and formate dehydrogenase.…”

Section: Construction Of a Multi-enzymatic Cascade Systemmentioning

confidence: 92%

“…Additionally, the catalytic activity of the E. coli pET-28a-laad-dapdh-fdh whole-cell biocatalyst was low (catalyzing 30 mM l-Phe), with only 1.5 mM concentration of the product, d-Phe. There is evidence that the closer a gene is to the end of a polycistronic operon, the lower is its expression [31]. Consequently, the first and the last positions in the polycistron would have the greatest impact on gene expression.…”

Section: Construction Of a Multi-enzymatic Cascade Systemmentioning

confidence: 99%

Highly selective synthesis of d-amino acids via stereoinversion of corresponding counterpart by an in vivo cascade cell factory

Zhang

Jing

et al. 2021

Microb Cell Fact

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Background d-Amino acids are increasingly used as building blocks to produce pharmaceuticals and fine chemicals. However, establishing a universal biocatalyst for the general synthesis of d-amino acids from cheap and readily available precursors with few by-products is challenging. In this study, we developed an efficient in vivo biocatalysis system for the synthesis of d-amino acids from l-amino acids by the co-expression of membrane-associated l-amino acid deaminase obtained from Proteus mirabilis (LAAD), meso-diaminopimelate dehydrogenases obtained from Symbiobacterium thermophilum (DAPDH), and formate dehydrogenase obtained from Burkholderia stabilis (FDH), in recombinant Escherichia coli. Results To generate the in vivo cascade system, three strategies were evaluated to regulate enzyme expression levels, including single-plasmid co-expression, double-plasmid co-expression, and double-plasmid MBP-fused co-expression. The double-plasmid MBP-fused co-expression strain Escherichia coli pET-21b-MBP-laad/pET-28a-dapdh-fdh, exhibiting high catalytic efficiency, was selected. Under optimal conditions, 75 mg/mL of E. coli pET-21b-MBP-laad/pET-28a-dapdh-fdh whole-cell biocatalyst asymmetrically catalyzed the stereoinversion of 150 mM l-Phe to d-Phe, with quantitative yields of over 99% ee in 24 h, by the addition of 15 mM NADP+ and 300 mM ammonium formate. In addition, the whole-cell biocatalyst was used to successfully stereoinvert a variety of aromatic and aliphatic l-amino acids to their corresponding d-amino acids. Conclusions The newly constructed in vivo cascade biocatalysis system was effective for the highly selective synthesis of d-amino acids via stereoinversion.

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“…d-PLA is an excellent biological preservative, while (R)-2-hydroxy-4-phenylbutyric acid is the key chiral intermediate for the production of angiotensin-converting enzyme inhibitors such as spirapril, enalaprilat, and lisinopril, which are widely used in the treatment of high blood pressure and congestive heart failure (Wang et al 2018a, b). D-danshensu has several pharmaceutical effects, including antitumor, antioxidant, antiinflammatory activity and efficacy in the prevention of myocardial ischemia (Xiong et al 2019). Furthermore, it can improve the overall heart function (Huo et al 2017).…”

Section: Introductionmentioning

confidence: 99%

Enzymological characterization of a novel d-lactate dehydrogenase from Lactobacillus rossiae and its application in d-phenyllactic acid synthesis

et al. 2020

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A novel lactate dehydrogenase gene, named lrldh, was cloned from Lactobacillus rossiae and heterologously expressed in Escherichia coli. The lactate dehydrogenase LrLDH is NADH-dependent with a molecular weight of approximately 39 kDa. It is active at 40 °C and pH 6.5 and stable in a neutral to alkaline environment below 35 °C. The kinetic constants, including maximal reaction rate (V max), apparent Michaelis-Menten constant (K m), turnover number (K cat) and catalytic efficiency (K cat /K m) for phenylpyruvic acid were 1.95 U mg −1 , 2.83 mM, 12.29 s −1 , and 4.34 mM −1 s −1 , respectively. Using whole cells of recombinant E. coli/pET28a-lrldh, without coexpression of a cofactor regeneration system, 20.5 g l −1 d-phenyllactic acid with ee above 99% was produced from phenylpyruvic acid in a fed-batch biotransformation process, with a productivity of 49.2 g l −1 d −1. Moreover, LrLDH has broad substrate specificity to a range of ketones, keto acids and ketonic esters. Taken together, LrLDH is a promising biocatalyst for the efficient synthesis of d-phenyllactic acid and other fine chemicals.

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One-pot, three-step cascade synthesis of D-danshensu using engineered Escherichia coli whole cells

Cited by 13 publications

References 26 publications

A Thermophilic Biofunctional Multienzyme Cascade Reaction for Cell-Free Synthesis of Salvianic Acid A and 3,4-Dihydroxymandelic Acid

A Thermophilic Biofunctional Multienzyme Cascade Reaction for Cell-Free Synthesis of Salvianic Acid A and 3,4-Dihydroxymandelic Acid

Highly selective synthesis of d-amino acids via stereoinversion of corresponding counterpart by an in vivo cascade cell factory

Enzymological characterization of a novel d-lactate dehydrogenase from Lactobacillus rossiae and its application in d-phenyllactic acid synthesis

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