Nitrilase immobilization and transposition from a micro‐scale batch to a continuous process increase the nicotinic acid productivity

Abstract

In recent years, many biocatalytic processes have been developed for the production of chemicals and pharmaceuticals. In this context, enzyme immobilization methods have attracted attention for their advantages, such as continuous production and increased stability. Here, enzyme immobilization methods and a collection of nitrilases from biodiversity for the conversion of 3‐cyanopyridine to nicotinic acid were screened. Substrate conversion over 10 conversion cycles was monitored to optimize the process. The best immobilization conditions were found with cross‐linking using glutaraldehyde to modify the PMMA beads. This method showed good activity over 10 cycles in a batch reactor at 30 and 40°C. Finally, production with a new thermostable nitrilase was examined in a continuous packed bed reactor, showing very high stability of the biocatalytic process at a flow rate of 0.12 ml min–1 and a temperature of 50°C. The complete conversion of 3‐cyanopyridine was obtained over 30 days of operation. Future steps will concern reactor scale‐up to increase the production rate with reasonable pressure drops.

Engineering of reaction specificity, enantioselectivity, and catalytic activity of nitrilase for highly efficient synthesis of pregabalin precursor

“…Nitrilases (EC 3.5.5.1) catalyze the hydrolysis of nitriles to the corresponding carboxylic acid and ammonia (Shen et al, 2020). Over the past decades, nitrilase‐mediated bioprocess has attracted much attention for manufacturing value‐added carboxylic acids (Desantis et al, 2003; Vergne‐Vaxelaire et al, 2013; Xue et al, 2015), including the building blocks of pharmaceuticals (Yu et al, 2020), pesticides (Dai et al, 2022), feed additives (Gong et al, 2018; Teepakorn et al, 2021) and polymers (Ben‐Bassat et al, 2008). The nitrilase‐catalyzed chemoenzymatic route for pregabalin began with the condensation of isovaleraldehyde and cyanoacetate, followed by cyanation to give isobutylsuccinonitrile (IBSN).…”

Microfluidic fabrication of tunable alginate‐based microfibers for the stable immobilization of enzymes

“…[8][9][10][11] The exploration of appropriate supporting matrix and novel technologies for enzyme immobilization has attracted intensive attentions. [8,[12][13][14][15][16][17][18][19][20][21][22] Microfluidic technique is a promising approach to manufacturing customizable structures by integrating with rapid crosslinking strategies. [18,21,[23][24][25][26][27] The convergence of the emerging microfluidic technology and materials science enables novel applications in various fields including enzyme immobilization.…”

“…[18,21,[23][24][25][26][27] The convergence of the emerging microfluidic technology and materials science enables novel applications in various fields including enzyme immobilization. [12,13,21] Alginate is a natural biomaterial widely used in the encapsulation of proteins/enzymes, drugs, and cells due to the capability of rapid crosslinking by multivalent cations (e.g., Ca 2+ or Ba 2+ ). [28][29][30][31][32][33][34] Alginate-based microfibers and microparticles can be feasibly fabricated via microfluidics technique which are widely used in fundamental researches and practical applications.…”

“…[ 8–11 ] The exploration of appropriate supporting matrix and novel technologies for enzyme immobilization has attracted intensive attentions. [ 8,12–22 ]…”

“…[8,[12][13][14][15][16][17][18][19][20][21][22] Microfluidic technique is a promising approach to manufacturing customizable structures by integrating with rapid crosslinking strategies. [18,21,[23][24][25][26][27] The convergence of the emerging microfluidic technology and materials science enables novel applications in various fields including enzyme immobilization. [12,13,21] Alginate is a natural biomaterial widely used in the encapsulation of proteins/enzymes, drugs, and cells due to the capability of rapid crosslinking by multivalent cations (e.g., Ca 2+ or Ba 2+ ).…”