A Regioselective Biocatalyst for Alkane Activation under Mild Conditions

Bordeaux, Mélanie; Galarneau, Anne; Fajula, François; Drone, Jullien

doi:10.1002/anie.201005597

Cited by 47 publications

(40 citation statements)

References 41 publications

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“…An alternative to the yeast CYP52 is the bacterial CYP153 which has been shown to hydroxylate short and medium chain fatty acids, alkanes, primary alcohols, and the terpene limonene (Bordeaux et al 2011;Scheps et al 2011Scheps et al , 2013Malca et al 2012;Cornelissen et al 2013). This three-component class I system consists of a heme-containing monooxygenase, an iron-sulfur ferredoxin, and a ferredoxin reductase domain (Bernhardt 2006).…”

Section: Introductionmentioning

confidence: 96%

“…In another study, the same three-component CYP153 system was expressed in E. coli and applied for production of octanol in a resting cell transformation at 1 mL scale leading to a final product concentration of 8.7 g L −1 after 24 h (Gudiminchi et al 2012) and 330 mg L −1 of 1-octanol was formed using P. putida (Vallon et al 2013). Furthermore, a fusion construct with CYP153A13a fused to the reductase domain of P450RhF expressed in E. coli was previously demonstrated to hydroxylate n-octane to 1-octanol at a 5-mL scale, however, not exceeding the product concentration above (Bordeaux et al 2011).…”

Section: Introductionmentioning

confidence: 98%

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Process limitations of a whole-cell P450 catalyzed reaction using a CYP153A-CPR fusion construct expressed in Escherichia coli

Lundemo

Notonier

Striedner

et al. 2015

Appl Microbiol Biotechnol

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Cytochrome P450s are interesting biocatalysts due to their ability to hydroxylate non-activated hydrocarbons in a selective manner. However, to date only a few P450-catalyzed processes have been implemented in industry due to the difficulty of developing economically feasible processes. In this study, we have used the CYP153A heme domain from Marinobacter aquaeolei fused to the reductase domain of CYP102A1 from Bacillus megaterium (BM3) expressed in Escherichia coli. This self-sufficient protein chimera CYP153A-CPRBM3 G307A mutant is able to selectively hydroxylate medium and long chain length fatty acids at the terminal position. ω-Hydroxylated fatty acids can be used in the field of high-end polymers and in the cosmetic and fragrance industry. Here, we have identified the limitations for implementation of a whole-cell P450-catalyzed reaction by characterizing the chosen biocatalyst as well as the reaction system. Despite a well-studied whole-cell P450 catalyst, low activity and poor stability of the artificial fusion construct are the main identified limitations to reach sufficient biocatalyst yield (mass of product/mass of biocatalyst) and space-time yield (volumetric productivity) essential for an economically feasible process. Substrate and product inhibition are also challenges that need to be addressed, and the application of solid substrate is shown to be a promising option to improve the process.

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Section: Introductionmentioning

confidence: 96%

Section: Introductionmentioning

confidence: 98%

Process limitations of a whole-cell P450 catalyzed reaction using a CYP153A-CPR fusion construct expressed in Escherichia coli

Lundemo

Notonier

Striedner

et al. 2015

Appl Microbiol Biotechnol

View full text Add to dashboard Cite

show abstract

“…Whole cell biotransformation can be used with toxic or insoluble substrates and products, without the supplementation of co-factor or additional substrate (Duetz et al, 2001;Li et al, 2002) and has been used for the efficient conversion of hydrophobic substrates such as fatty acids (Joo et al, 2012b;Julsing et al, 2012;Kishimoto et al, 2003), carboxylic acids ( (Bordeaux et al, 2011) to oxygenated products. Not only several bacteria (Hudson et al, 1998;Kishimoto et al, 2003;Takeuchi et al, 2013), but also recombinant E. coli cells expressing linoleate 13-hydratase (Oh et al, 2015) have produced 13-HOD.…”

Section: Discussionmentioning

confidence: 99%

Production of 13S-hydroxy-9(Z)-octadecenoic acid from linoleic acid by whole recombinant cells expressing linoleate 13-hydratase from Lactobacillus acidophilus

Park

Lee

Kim

et al. 2015

Journal of Biotechnology

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“…Optimization of the flux of electrons is crucial to utilize the full capacity of the biocatalyst to the targeted reaction, and the importance has also been emphasized in the review by Bernhardt and Urlacher (2014). Approaches with fusion constructs have been published, and besides constructs with the reductase domain of BM3, the reductase domain of CYP116B2 (P450RhF) is a promising alternative (Bordeaux et al 2011;Robin et al 2009). This field with a lot of potential for the application of P450s has recently been reviewed (Sadeghi and Gilardi 2013).…”

Section: Electron Transport and Coupling Efficiencymentioning

confidence: 97%

Guidelines for development and implementation of biocatalytic P450 processes

Lundemo

Woodley

2015

Appl Microbiol Biotechnol

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Biocatalytic reactions performed by cytochrome P450 monooxygenases are interesting in pharmaceutical research since they are involved in human drug metabolism. Furthermore, they are potentially interesting as biocatalysts for synthetic chemistry because of the exquisite selectivity of the chemistry they undertake. For example, selective hydroxylation can be undertaken on a highly functionalized molecule without the need for functional group protection. Recent progress in the discovery of novel P450s as well as protein engineering of these enzymes strongly encourages further development of their application, including use in synthetic processes. The biological characteristics of P450s (e.g., cofactor dependence) motivate the use of whole-cell systems for synthetic processes, and those processes implemented in industry are so far dominated by growing cells and native host systems. However, for an economically feasible process, the expression of P450 systems in a heterologous host with sufficient biocatalyst yield (g/g cdw) for non-growing systems or space-time yield (g/L/h) for growing systems remains a major challenge. This review summarizes the opportunities to improve P450 whole-cell processes and strategies in order to apply and implement them in industrial processes, both from a biological and process perspective. Indeed, a combined approach of host selection and cell engineering, integrated with process engineering, is suggested as the most effective route to implementation.

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A Regioselective Biocatalyst for Alkane Activation under Mild Conditions

Cited by 47 publications

References 41 publications

Process limitations of a whole-cell P450 catalyzed reaction using a CYP153A-CPR fusion construct expressed in Escherichia coli

Process limitations of a whole-cell P450 catalyzed reaction using a CYP153A-CPR fusion construct expressed in Escherichia coli

Production of 13S-hydroxy-9(Z)-octadecenoic acid from linoleic acid by whole recombinant cells expressing linoleate 13-hydratase from Lactobacillus acidophilus

Guidelines for development and implementation of biocatalytic P450 processes

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