Cyanide-free synthesis of an aromatic nitrile from a biorenewable-based aldoxime: Development and application of a recombinant aldoxime dehydratase as a biocatalyst

Choi, Ji Eun; Shinoda, Suguru; Inoue, Ryo; Zheng, Daijun; Gröger, Harald; Asano, Yasuhisa

doi:10.1080/10242422.2019.1591376

Cited by 15 publications

(27 citation statements)

References 20 publications

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“…The clones that showed an absorbance between 0 and 1 (lower 10%) at 270 nm ( Figure 1) were chosen for further studies. In continuation of our research on the biocatalytic synthesis of nitriles under cyanide-free conditions as an alternative synthetic technology for nitriles [13][14][15][16][17][18][19][20][21][22][23][24][25], we could also demonstrate a proof of concept for the two-step conversion of furfural into 2-furonitrile (Scheme 1) [24,25]. In detail, furfural (1) is transformed into its aldoxime (2) by spontaneous condensation with hydroxylamine, which is a bulk chemical that is utilized in α-caprolactam manufacturing.…”

Section: High Throughput Screening Of Oxdyh3-3 Mutants By Using a Uv mentioning

confidence: 99%

“…In the next steps, we gained interest in evaluating the activity of selected mutants in synthetic biotransformations. As in our previous work, the substrate concentration with the wild-type enzyme was only 10 mM [25]. We studied these mutants in the dehydration of furfuryl aldoxime at increased 50 and 100 mM concentrations in the presence of a biocatalyst loading of approximately 0.5 g•L -1 wet cells ( Table 1).…”

Section: The Improved Activity Measurement Of Oxdyh3-3 Mutantsmentioning

confidence: 99%

“…We studied these mutants in the dehydration of furfuryl aldoxime at increased 50 and 100 mM concentrations in the presence of a biocatalyst loading of approximately 0.5 g•L -1 wet cells ( Table 1). At first, E-3-pyridine aldoxime was used as a standard substrate since OxdYH3-3 showed the highest relative activity for E-3-pyridine aldoxime in a previous study [24,25]. At an increased substrate concentration of 100 mM of E-3-pyridine aldoxime, OxdYH3-3 N266S showed a three times higher activity (1.73 U•g -1 ; measured according to the conversion after 1 h) compared to the one of the wild-type enzyme (0.52 U•g -1 ; Table 1).…”

Section: The Improved Activity Measurement Of Oxdyh3-3 Mutantsmentioning

confidence: 99%

“…Chemicals including E-3-pyridine aldoxime, 3-cyanopyridine, 2-furaldehyde, and 2-furonitrile were purchased from Sigma Aldrich (Germany). OxdYH3-3 WT in a pET28b vector was constructed and transformed into BL21(DE3) Escherichia coli cell followed by methods in the previous study [25].…”

Section: Materials and Preparation Of Oxdyh3-3 Transformantsmentioning

confidence: 99%

“…However, in spite of the availability of a recombinant form of an aldoxime dehydratase from Rhodococcus sp. YH3-3 (OxdYH3-3) [25], which was identified as a suitable biocatalyst for aromatic aldoximes in general and 2-furonitrile in particular, the limited enzyme activity and substrate concentration raise the demand for more active and stable biocatalysts as well as for a process running at elevated substrate loading. In the following, we report our results on the development of improved mutants of this enzyme that show an increased activity and enable an improved synthetic access to 2-furonitrile.…”

Section: Introductionmentioning

confidence: 99%

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Aldoxime Dehydratase Mutants as Improved Biocatalysts for a Sustainable Synthesis of Biorenewables-Based 2-Furonitrile

et al. 2020

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2-Furonitrile is an interesting nitrile product for the chemical industry due to its use as intermediate in the field of fine chemicals and pharmaceuticals or as a potential sweetener, as well as due to its access from biorenewables. As an alternative to current processes based on, e.g., the ammoxidation of furfural with ammonia as a gas phase reaction running at > 400 °C, we recently reported an enzymatic dehydration of 2-furfuryl aldoxime being obtained easily from furfural and hydroxylamine. However, improving the catalytic properties of the aldoxime dehydratase biocatalyst from Rhodococcus sp. YH3-3 (OxdYH3-3) in terms of activity and stability remained a challenge. In this contribution, the successful development of aldoxime dehydratase OxdYH3-3 mutants that were generated by directed evolution and its enhanced activity toward 2-furfuryl aldoxime is reported. The mutant OxdYH3-3 N266S showed an improved activity of up to six times higher than the wild type when utilizing a substrate concentration of 50–100 mM of 2-furfuryl aldoxime.

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Section: High Throughput Screening Of Oxdyh3-3 Mutants By Using a Uv mentioning

confidence: 99%

Section: The Improved Activity Measurement Of Oxdyh3-3 Mutantsmentioning

confidence: 99%

Section: The Improved Activity Measurement Of Oxdyh3-3 Mutantsmentioning

confidence: 99%

Section: Materials and Preparation Of Oxdyh3-3 Transformantsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Aldoxime Dehydratase Mutants as Improved Biocatalysts for a Sustainable Synthesis of Biorenewables-Based 2-Furonitrile

et al. 2020

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Synthetic Processes toward Nitriles without the Use of Cyanide: A Biocatalytic Concept Based on Dehydration of Aldoximes in Water

Hinzmann

Betke

Asano

et al. 2021

Chemistry A European J

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While belonging to the mostf undamentalf unctional groups,n itrilesr epresent ac lass of compound that still raises challenges in terms of an efficient, cost-effective, generala nd,a tt he same time, sustainable way for their synthesis. Complementing existing chemical routes, recently ac yanide-free enzymatic process technology based on the use of an aldoximed ehydratase (Oxd) as a biocatalyst component has been developed and successfully applied fort he synthesis of ar ange of nitrile products. In these biotransformations,t he Oxd enzymes catalyze the dehydration of aldoximes as readily available substrates to the nitrile products.H erein, these developments with such enzymesa re summarized, with as trong focus on synthetic applications. It is demonstrated that this biocatalytic technology has the potential to "cross the bridge" between the production of fine chemicals and pharmaceuticals, on one hand, and bulk and commodity chemicals, on the other.

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Selective Biocatalytic Defunctionalization of Raw Materials

Wohlgemuth

2022

ChemSusChem

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Biobased raw materials, such as carbohydrates, amino acids, nucleotides, or lipids contain valuable functional groups with oxygen and nitrogen atoms. An abundance of many functional groups of the same type, such as primary or secondary hydroxy groups in carbohydrates, however, limits the synthetic usefulness if similar reactivities cannot be differentiated. Therefore, selective defunctionalization of highly functionalized biobased starting materials to differentially functionalized compounds can provide a sustainable access to chiral synthons, even in case of products with fewer functional groups. Selective defunctionalization reactions, without affecting other functional groups of the same type, are of fundamental interest for biocatalytic reactions. Controlled biocatalytic defunctionalizations of biobased raw materials are attractive for obtaining valuable platform chemicals and building blocks. The biocatalytic removal of functional groups, an important feature of natural metabolic pathways, can also be utilized in a systemic strategy for sustainable metabolite synthesis.

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Cyanide-free synthesis of an aromatic nitrile from a biorenewable-based aldoxime: Development and application of a recombinant aldoxime dehydratase as a biocatalyst

Cited by 15 publications

References 20 publications

Aldoxime Dehydratase Mutants as Improved Biocatalysts for a Sustainable Synthesis of Biorenewables-Based 2-Furonitrile

Aldoxime Dehydratase Mutants as Improved Biocatalysts for a Sustainable Synthesis of Biorenewables-Based 2-Furonitrile

Synthetic Processes toward Nitriles without the Use of Cyanide: A Biocatalytic Concept Based on Dehydration of Aldoximes in Water

Selective Biocatalytic Defunctionalization of Raw Materials

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