Medium-Scale Preparation of Useful Metabolites of Aromatic Compounds via Whole-Cell Fermentation with Recombinant Organisms

Endoma, Mary Ann A.; Bui, Vu; Hansen, Jeff; Hudlický, Tomáš

doi:10.1021/op020013s

Cited by 91 publications

(60 citation statements)

References 65 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This transformation is remarkable since it accomplishes dearomatization of benzene derivatives to give arene dihydrodiols, allowing for synthetic routes, which would otherwise not be feasible. It is also a whole-cell fermentation (in E. coli) which appears well-scalable (up to 15 l of broth in a laboratory setting), furnishing the dihydrodiols on 10 to 100 g scale within three days, including the time needed for preculture preparation and inoculation (Endoma et al, 2002). Compounds of this sort, the production of which by chemical means would be a lengthy procedure, are highly useful building blocks because chemical synthesis might be next used to elaborate them further to plant compounds such as pancratistatin and morphine (Hudlicky, 2006;Reed and Hudlicky, 2015), or analogs and derivatives thereof, and also oseltamivir (Werner et al, 2010), in the order of ten steps.…”

Section: Organic Synthesismentioning

confidence: 99%

Discovery and resupply of pharmacologically active plant-derived natural products: A review

2015

Draw Science

View full text Add to dashboard Cite

Medicinal plants have historically proven their value as a source of molecules with therapeutic potential, and nowadays still represent an important pool for the identification of novel drug leads. In the past decades, pharmaceutical industry focused mainly on libraries of synthetic compounds as drug discovery source. They are comparably easy to produce and resupply, and demonstrate good compatibility with established high throughput screening (HTS) platforms. However, at the same time there has been a declining trend in the number of new drugs reaching the market, raising renewed scientific interest in drug discovery from natural sources, despite of its known challenges. In this survey, a brief outline of historical development is provided together with a comprehensive overview of used approaches and recent developments relevant to plant-derived natural product drug discovery. Associated challenges and major strengths of natural product- Europe PMC Funders Author ManuscriptsEurope PMC Funders Author Manuscripts based drug discovery are critically discussed. A snapshot of the advanced plant-derived natural products that are currently in actively recruiting clinical trials is also presented. Importantly, the transition of a natural compound from a "screening hit" through a "drug lead" to a "marketed drug" is associated with increasingly challenging demands for compound amount, which often cannot be met by re-isolation from the respective plant sources. In this regard, existing alternatives for resupply are also discussed, including different biotechnology approaches and total organic synthesis.While the intrinsic complexity of natural product-based drug discovery necessitates highly integrated interdisciplinary approaches, the reviewed scientific developments, recent technological advances, and research trends clearly indicate that natural products will be among the most important sources of new drugs also in the future.

show abstract

Section: Organic Synthesismentioning

confidence: 99%

Discovery and resupply of pharmacologically active plant-derived natural products: A review

2015

Draw Science

View full text Add to dashboard Cite

show abstract

“…Here, continuous phase extraction with ethyl acetate might be used as an alternative to the evaporation of the aqueous phase. [32] Table 1 lists the process parameters of our cinnamonitrile biotransformation using E. coli JM101 (pTEZ30) on different scales. The downstream processing and the purification yields with the corresponding purities are given in Table 2.…”

Section: Biotransformation Of Cinnamonitrile On a 30 L-scalementioning

confidence: 99%

“…Diol 2 was obtained from the whole cell fermentation of bromobenzene with E. coli JM109 (pDTG 601), a strain developed by David Gibson and expressing the toluene dioxygenase genes. [12,32] This material was also reduced with PAD and the diol protected as acetonide 5.…”

Section: Proof Of Absolute Stereochemistrymentioning

confidence: 99%

“…[28 ± 31] Biotransformations with recombinant cells expressing dioxygenase genes were performed on laboratory and industrial production scale yielding gram to multi-kilogram amounts of cis-cyclohexadiene diols. [32] In general, aromatic nitriles are interesting synthons, because the enzymatic dihydroxylation may be coupled to a selective hydrolysis of the nitrile group or subse- quent cycloaddition reactions to increase the pool of chiral synthons. We now examine the efficiency of a bioprocess for the highly selective asymmetric dihydroxylation of non-natural cinnamonitrile to trans-3-[(5S,6R)-5,6-dihydroxycyclohexa-1,3-dienyl]-acrylonitrile (1) by chlorobenzene dioxygenase (CDO) in recombinant E. coli JM101 (pTEZ30).…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Asymmetric Dihydroxylation of Cinnamonitrile to trans‐3‐[(5S,6R)‐5,6‐Dihydroxycyclohexa‐1,3‐dienyl]‐acrylonitrile using Chlorobenzene Dioxygenase in Escherichia coli (pTEZ30)

et al. 2004

Self Cite

View full text Add to dashboard Cite

Asymmetric cis-dihydroxylations of aromatic compounds are catalyzed by bacterial dioxygenases. In order to prevent through conversion, either dihydrodiol dehydrogenase blocked mutant strains or recombinant bacterial cells are used as biocatalysts for synthetic purposes. We characterized the cis-dihydroxylation of cinnamonitrile by chlorobenzene dioxygenase (CDO) in recombinant E. coli on different reaction scales. The absolute stereochemistry of the product was determined to be trans-3-[(5S,6R)-5,6-dihydroxycyclohexa-1,3-dienyl]-acrylonitrile. The cells showed a maximum specific activity of 3.76 U/g cdw in shake-flask experiments. Stable expression of the dioxygenase genes in E. coli JM101 (pTEZ30) resulted in increasing volumetric productivities. The maximum volumetric productivities of 80 and 92 mg product/L/h were achieved on 2-L and 30-L scales, respectively. The specific growth rate correlated with the volumetric productivity during the biotransformations. An average volumetric productivity of 40 mg product/L/h in reactors on 2-L and 30-L scales resulted in 0.96 and 16.4 g of isolated product at the end of the biotransformations. This points out the need for metabolically active cells and controllable expression systems for achieving high volumetric productivities for cofactor dependent biooxidations. We have now applied this concept for the asymmetric dihydroxylation of the non-natural substrate cinnamonitrile using multicomponent CDO in tailored E. coli JM101 in long-term reactions.

show abstract

“…8 The application of these in asymmetric synthesis of complex molecules are numerous and have been reviewed on several occasions. [9][10][11] Examples of the diversity of metabolites produced substrates that are tolerated and metabolized by TDO are shown in Figure 1.…”

mentioning

confidence: 99%

Chemoenzymatic synthesis of complex natural and unnatural products: morphine, pancratistatin, and their analogs

Hudlický¹

2006

Arkivoc

Self Cite

View full text Add to dashboard Cite

The synthesis of morphine (1) and pancratistatin (2) have been the subject of intense effort by the synthetic community for many years. Our focus on the total synthesis of these challenging targets resulted in several generations of approaches that combine enzymatic transformations with traditional synthetic protocols in order to provide for maximum efficiency and brevity in attaining the targets.Recombinant strains that express toluene dioxygenase (TDO) are used to provide the homochiral diene cis-diol derived from bromobenzene and containing the structural features of both ring C of morphine and ring C of pancratistatin. Bromocatechol, representing ring A of morphine, is also derived from bromobenzene by fermentation with E.coli JM109 that expresses TDO as well as catechol dehydrogenase. In this fashion twelve of the carbons in morphine originate in the same starting material. One approach is based on the Kazmaier-Claisen rearrangement of glycinate esters, the other on an intramolecular Heck reaction. The route to pancratistatin and to its unnatural analogs is based on the metal-mediated cyclotrimerization of acetylenic substrates of type 3. The core of this Amaryllidaceae constituent has been attained in the initial stages of this project. The details of the synthetic endeavours toward these and other heterocyclic targets will be disclosed with emphasis on practicality, brevity, and efficiency as guiding principles of enviromentally benign manufacturing of relevant compounds.

show abstract

Medium-Scale Preparation of Useful Metabolites of Aromatic Compounds via Whole-Cell Fermentation with Recombinant Organisms

Cited by 91 publications

References 65 publications

Discovery and resupply of pharmacologically active plant-derived natural products: A review

Discovery and resupply of pharmacologically active plant-derived natural products: A review

Asymmetric Dihydroxylation of Cinnamonitrile to trans‐3‐[(5S,6R)‐5,6‐Dihydroxycyclohexa‐1,3‐dienyl]‐acrylonitrile using Chlorobenzene Dioxygenase in Escherichia coli (pTEZ30)

Chemoenzymatic synthesis of complex natural and unnatural products: morphine, pancratistatin, and their analogs

Contact Info

Product

Resources

About