High Regioselectivity Production of 5-Cyanovaleramide from Adiponitrile by a Novel Nitrile Hydratase Derived from <i>Rhodococcus erythropolis</i> CCM2595

Wang, Li; Liu, Shengxian; Du, Wenjing; Dou, Tingfeng; Liang, Changhai

doi:10.1021/acsomega.0c02188

Cited by 10 publications

(6 citation statements)

References 20 publications

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“…In our work, the main product was still 5-cyanovaleramide, though only a small amount of adipamide was produced (Figure 4) when treated with the purified ReNHase only within 2 min. In our previous study, the adiponitrile could completely be consumed within 10 min when it was treated with whole-cell catalysis [16]. The amount of 5cyanovaleramide decreased and the amount of adipamide increased in a time-dependent manner.…”

Section: Discussionmentioning

confidence: 86%

“…Significantly, the purified ReNHase exhibited higher activity during the conversion reaction of different nitriles by shortened catalytic time and increased yield of monocyanate products. In our previous study, during the whole-cell catalysis, 20 mmol L −1 adiponitrile, malononitrile, 2 mmol L −1 terephthalonitrile(after ultrasound, the mixture was homogeneous) could be completely transformed within 10 min, 3 h and 30 min, respectively [16].…”

Section: Discussionmentioning

confidence: 97%

“…In our previous study [16], ReNHase of Rhodococcus. erythropolis CCM2595 manifested high biocatalytic activity.…”

Section: Introductionmentioning

confidence: 91%

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Efficient biodegradation of nitriles by a novel nitrile hydratase derived from Rhodococcus erythropolis CCM2595

Huang

Cui

et al. 2021

Biotechnology & Biotechnological Equipment

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In our previous study, we firstly cloned the gene of a novel ReNHase (NHase from Rhodococcus erythropolis CCM2595), and the strain was shown to degrade only phenol, hydroxybenzoate, p-chlorophenol, aniline and other aromatic compounds. Here in, we further purified the ReNHase from R. erythropolis CCM2595 and detected its properties of biodegradation. We constructed a plasmid with the gene of ReNHase with His-tag. The encoding ReNHase was cloned and overexpressed in recombinant Escherihia coli and confirmed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The substrate scope of the new heterologous ReNHase was tested and the efficiency of degradation of nitriles by the ReNHase was also investigated by high efficiency liquid chromatography. We also studied the effect of temperature and pH on the catalysis of adiponitrile by the purified ReNHase. The recombinant E.coli showed high catalytic regioselectivity with high substrate affinity towards dinitriles (especially for adiponitrile) whereas lower affinity towards mononitriles. Compared to whole-cell catalysis, the catalytic time was shortened significantly with the purified ReNHase. The enzyme activity of crude recombinant E.coli was 635 U g −1 (DCW), while the specific activity of purified ReNHase was 63.107 U mg −1 . The apparent Km value for the purified ReNHase is 6.6252 mmol L −1 , which revealed the good affinity between the purified ReNHase and adiponitrile. The reaction Kcat is 82.77 s −1 and Kcat/Km is 1.249 × 10 4 (mol −1 L s −1 ), which showed high catalytic activity towards adiponitrile. We propose that this purified ReNHase may be applied for the industry and sewage treatment for environmental protection.

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Section: Discussionmentioning

confidence: 86%

Section: Discussionmentioning

confidence: 97%

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Efficient biodegradation of nitriles by a novel nitrile hydratase derived from Rhodococcus erythropolis CCM2595

Huang

Cui

et al. 2021

Biotechnology & Biotechnological Equipment

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“…Rhodococcus can survive in hostile environments, such as those contaminated with polychlorinated biphenyl [30], nitrophenols, trichloroethene [31], and polycyclic aromatic hydrocarbons [32]. Studies have shown that Rhodococcus can degrade and remove aromatic [33], nitrile [34], sulfuric [35], and nitrogenous compounds [36] in the environment. Indeed, Rhodococcus are commonly used to remediate environmental contamination and pollution [37].…”

Section: Discussionmentioning

confidence: 99%

Effects of Cadmium Stress on Bacterial and Fungal Communities in the Whitefly Bemisia tabaci

Guo,

Li,

2023

IJMS

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Heavy metal contamination is among the most prominent environmental problems in China, posing serious threats to both ecosystem and human health. Among the diverse heavy metal contaminants, cadmium is the most serious. The whitefly Bemisia tabaci is a cosmopolitan pest capable of causing severe damage to a broad range of agricultural crops, especially vegetables. At present, little is known about the effects of cadmium stress on B. tabaci, including on its bacterial and fungal communities. In the current study, we investigated the effects of cadmium on bacterial and fungal communities in whiteflies. Meta-barcode sequencing of the 16S rRNA gene revealed that the whitefly bacterial community contained 264 operational taxonomic units (OTUs) belonging to 201 known genera and 245 known species. The top five most frequent bacterial genera were Rickettsia, Rhodococcus, Candidatus Portiera, Candidatus Hamiltonella, and Achromobacter. Meta-barcode sequencing of the fungal ITS locus revealed that the whitefly fungal community contained 357 OTUs belonging to 187 known genera and 248 known species. The top five most frequent fungal genera were Wallemia, unclassified_f_Dipodascaceae, Apiotrichum, Penicillium, and unclassified_o_Saccharomycetales. Cadmium exposure reduced the fungal OTU richness but increased the bacterial Shannon and Simpson diversity indices in whiteflies. In addition, upon exposure to cadmium, the microbial community composition in whiteflies changed significantly, with increased prevalence of the bacterial genera Rhodococcus and Exiguobacterium and fungal genus Wallemia. Our results indicate that the whitefly microbiota likely contributed to their adaptation and resistance to cadmium and suggested that whiteflies may contain microbes that could help remediate cadmium contamination in natural environments and agricultural fields.

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“…48−50 To combat these issues, protein engineering has been applied to enhance expression, simplify purification, and expand substrate stability and range for various wild NHases. 51,52 Additionally, NHase immobilization is vital for large-scale chemical production, although structural instability and methods like glutaraldehyde cross-linking or MOFs encapsulation may reduce its activity. 53−55 In this study, we introduce a "preblocking" method utilizing VS groups to control NHase immobilization and its surrounding hydrophobic microenvironment.…”

Section: ■ Introductionmentioning

confidence: 99%

A Preblocking Strategy with Rapid Immobilization of Nitrile Hydratase and Precise Control of Hydrophobic Microenvironment for High Regioselective Amide Transformation

Dong,

Wu,

Weng

et al. 2024

ACS Sustainable Chem. Eng.

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The control over enzyme immobilization and its microenvironment at the molecular level is deemed crucial. In this study, a "preblocking" method was performed through the use of natural polymer agarose resins after activation by phenyl-, methyl-, and ethyl-vinyl sulfones combined with divinyl sulfones prior to the specific immobilization of a his-tag fused recombinant nitrile hydratase (ReNHase). This technique primarily allows for the establishment of a hydrophobic microenvironment to enhance the immobilization and mass transfer and avoid inactivation caused by traditional backfilling. The purified ReNHase showed specific activity and regioselectivity of adiponitrile in an aqueous solution as 28.43 U•mg −1 and 90.2%, respectively. Increasing the density of hydrophobic groups promotes the adsorption extent and immobilization yield of ReNHase. Optimized DVS:PVS 1:4 hydrophobic microenvironment could finish immobilization in 4h. Eminent expressed activity and regioselectivity of 94.6% and 93.9% was exhibited, respectively. The approach demonstrated exceptional selectivity in the biotransformation at the molecular level. Furthermore, the method was found to preserve enzymatic activity over seven cycles and exhibited significantly enhanced resilience to variations in the temperature and substrate concentration. The ReNHase@DVS:PVS 1:4 still retained 98% of the maximum activity, while ReNHase only retained 64% of activity at 55 °C. The ReNHase@DVS:PVS 1:4 still retained 55% of the maximum activity, while ReNHase was completely inactive at 200 mM adiponitrile. A packed-bed reactor was constructed, where the biotransformation of 5.64 g of 5-cyanovaleramide was carried out continuously for 48 h at a flow rate of 0.60 mL/min, achieving a peak space-time yield reported to be 0.93 mol•h −1 •L −1 . The "preblocking" technique developed in this study is anticipated to provide new methodology in the catalysis of hydrophobic substrates.

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High Regioselectivity Production of 5-Cyanovaleramide from Adiponitrile by a Novel Nitrile Hydratase Derived from Rhodococcus erythropolis CCM2595

Cited by 10 publications

References 20 publications

Efficient biodegradation of nitriles by a novel nitrile hydratase derived from Rhodococcus erythropolis CCM2595

Efficient biodegradation of nitriles by a novel nitrile hydratase derived from Rhodococcus erythropolis CCM2595

Effects of Cadmium Stress on Bacterial and Fungal Communities in the Whitefly Bemisia tabaci

A Preblocking Strategy with Rapid Immobilization of Nitrile Hydratase and Precise Control of Hydrophobic Microenvironment for High Regioselective Amide Transformation

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