Characterization and immobilization of Pycnoporus cinnabarinus carboxylic acid reductase, PcCAR2

Maphatsoe, Masethabela Maria; Hashem, Chiam; Ling, Jonathan Guyang; Horvat, Melissa; Rumbold, Karl; Bakar, Farah Diba Abu; Winkler, Margit

doi:10.1016/j.jbiotec.2021.12.010

Cited by 8 publications

(8 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…GCD40661.1 1/21 [36] 53 MaCAR4 Mycobacteroides abscessus CAM63041.1 7/21 [37] 54 PcCAR2 Pycnoporus cinnabarinus UMO78114.1 12/ 21 [38] n.a. not available; [a] First published evidence of functional heterologous expression.…”

Section: Streptomyces Paromomycinusmentioning

confidence: 99%

Biocatalytic Carboxylate Reduction – Recent Advances and New Enzymes

Winkler

Ling

2022

ChemCatChem

Self Cite

View full text Add to dashboard Cite

Carboxylate reductases (CARs) are valuable catalysts for the selective one‐step reduction of carboxylic acids to their corresponding aldehydes. In recent years, numerous new CARs have been made available, studied and applied in the context of biocatalytic syntheses. The preparation of aldehydes as end products for the flavor and fragrance sector and the integration of CARs in cascade reactions with aldehydes as the key intermediates represent the two major fields of applications. This review gives a comprehensive overview of the current toolbox of recombinant CARs and their numerous carboxylate substrates. Non‐natural functions of CARs are highlighted and recent insight into the structure‐function relationship of CARs are summarized.

show abstract

Section: Streptomyces Paromomycinusmentioning

confidence: 99%

Biocatalytic Carboxylate Reduction – Recent Advances and New Enzymes

Winkler

Ling

2022

ChemCatChem

Self Cite

View full text Add to dashboard Cite

show abstract

“…However, some mechanistic and structural insight was deduced from crystallized single and didomains of bacterial CARs . CARs can be divided into five clans: The group of bacterial CARs belongs to type I, while type III consists of CARs from Ascomycetes, type IV of CARs from Basidiomycetes and type II represents a mixed group of fungal origin . The fifth type of enzymes of similar domain architecture is specialized in reducing aromatic amino acids .…”

mentioning

confidence: 99%

Structure of the Reductase Domain of a Fungal Carboxylic Acid Reductase and Its Substrate Scope in Thioester and Aldehyde Reduction

et al. 2022

Self Cite

View full text Add to dashboard Cite

The synthesis of aldehydes from carboxylic acids has long been a challenge in chemistry. In contrast to the harsh chemically driven reduction, enzymes such as carboxylic acid reductases (CARs) are considered appealing biocatalysts for aldehyde production. Although structures of single- and didomains of microbial CARs have been reported, to date no full-length protein structure has been elucidated. In this study, we aimed to obtain structural and functional information regarding the reductase (R) domain of a CAR from the fungus Neurospora crassa (Nc). The NcCAR R-domain revealed activity for N-acetylcysteamine thioester (S-(2-acetamidoethyl) benzothioate), which mimics the phosphopantetheinylacyl-intermediate and can be anticipated as the minimal substrate for thioester reduction by CARs. The determined crystal structure of the NcCAR R-domain reveals a tunnel that putatively harbors the phosphopantetheinylacyl-intermediate, which is in good agreement with docking experiments performed with the minimal substrate. In vitro studies were performed with this highly purified R-domain and NADPH, demonstrating carbonyl reduction activity. The R-domain was able to accept not only a simple aromatic ketone but also benzaldehyde and octanal, which are typically considered to be the final product of carboxylic acid reduction by CAR. Also, the full-length NcCAR reduced aldehydes to primary alcohols. In conclusion, aldehyde overreduction can no longer be attributed exclusively to the host background.

show abstract

“…It was observed that the immobilized Mp CAR retained 59.68% of its catalytic activity even after 10 times of usage. Likewise, immobilization of CAR from Pycnoporus cinnabarinus ( Pc CAR2) showed good reusability as it retained >80% of its initial activity after six cycles of activity assay [ 30 ]. Based on the Bradford assay conducted in this study, Mp CAR was not detected in the supernatant of the reaction mixture even after 10 cycles of the assay (data not shown).…”

Section: Resultsmentioning

confidence: 99%

“…A previous study revealed that ~50% of bioconversion of 5 mM of benzoic acid was achieved by immobilized CAR from Segniliparus rugosus ( Sr CAR) after 18 h of incubation at 30 °C [ 29 ]. Another finding showed that ~80% bioconversion yield was achieved by immobilized Pc CAR, given 2 mM of benzoic acid was supplied for 1 h of reaction incubation at 25 °C [ 30 ]. Here, the 2.6 mM of benzaldehyde was quantified after 1 h of incubation of immobilized Mp CAR at 60 °C, even though the yield was only about 50%.…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Characterization of Carboxylic Acid Reductase from Mycobacterium phlei Immobilized onto Seplite LX120

et al. 2022

View full text Add to dashboard Cite

A multi-domain oxidoreductase, carboxylic acid reductase (CAR), can catalyze the one-step reduction of carboxylic acid to aldehyde. This study aimed to immobilize bacterial CAR from a moderate thermophile Mycobacterium phlei (MpCAR). It was the first work reported on immobilizing bacterial CAR onto a polymeric support, Seplite LX120, via simple adsorption. Immobilization time and protein load were optimized for MpCAR immobilization. The immobilized MpCAR showed optimal activity at 60 °C and pH 9. It was stable over a wide range of temperatures (10 to 100 °C) and pHs (4–11), retaining more than 50% of its activity. The immobilized MpCAR also showed stability in polar solvents. The adsorption of MpCAR onto the support was confirmed by Scanning Electron Microscopy (SEM), Fourier-Transform Infrared (FTIR) spectroscopy, and Brunauer–Emmett–Teller (BET) analysis. The immobilized MpCAR could be stored for up to 6 weeks at 4 °C and 3 weeks at 25 °C. Immobilized MpCAR showed great operational stability, as 59.68% of its activity was preserved after 10 assay cycles. The immobilized MpCAR could also convert approximately 2.6 mM of benzoic acid to benzaldehyde at 60 °C. The successfully immobilized MpCAR on Seplite LX120 exhibited improved properties that benefit green industrial processes.

show abstract

Characterization and immobilization of Pycnoporus cinnabarinus carboxylic acid reductase, PcCAR2

Cited by 8 publications

References 42 publications

Biocatalytic Carboxylate Reduction – Recent Advances and New Enzymes

Biocatalytic Carboxylate Reduction – Recent Advances and New Enzymes

Structure of the Reductase Domain of a Fungal Carboxylic Acid Reductase and Its Substrate Scope in Thioester and Aldehyde Reduction

Characterization of Carboxylic Acid Reductase from Mycobacterium phlei Immobilized onto Seplite LX120

Contact Info

Product

Resources

About