Cytochrome <i>c</i>–poly(acrylic acid) conjugates with improved peroxidase turnover number

Benson, Kyle; Gorecki, J; Никифоров, А. К.; Tsui, William G.; Kasi, Rajeswari M.; Kumar, Challa V.

doi:10.1039/c9ob00541b

Cited by 11 publications

(16 citation statements)

References 33 publications

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“…When substrate concentration is excessive, k cat tends to V max . 26 Therefore, the k cat of free and immobilized enzymes was 0.060 and 0.040 s –1 , respectively, the k cat / K m of the free and immobilized pectinase was 0.005 and 0.003 mg –1 ·s –1 ·mL, respectively. V max , k cat , and k cat / K m decreased possibly because the substrate could not bind to the active sites of the immobilized pectinase because of the increased diffusion limitation.…”

Section: Resultsmentioning

confidence: 91%

Immobilization of Pectinase onto Porous Hydroxyapatite/Calcium Alginate Composite Beads for Improved Performance of Recycle

Gao

et al. 2020

ACS Omega

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Pectinase is an industrially important enzyme widely used in juice production, food processing, and other fields. The use of immobilized enzyme systems that allow several reuses of pectinase is beneficial to these fields. Herein, we developed mechanically strong and recyclable porous hydroxyapatite/calcium alginate composite beads for pectinase immobilization. Under the optimal immobilization parameters of 40 °C, pH 4.0, 5.2 U/L pectinase concentration and 4 h reaction time, pectinase showed the highest enzymatic activity (8995 U/mg) and immobilization yield (91%). The thermal stability and pH tolerance of the immobilized pectinase were superior to those of free pectinase. The storage stability of the free and immobilized pectinase for 30 days retained 20 and 50% of their initial activity, respectively. Therefore, these composite beads might be promising support for the efficient immobilization of industrially important enzymes.

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Section: Resultsmentioning

confidence: 91%

Immobilization of Pectinase onto Porous Hydroxyapatite/Calcium Alginate Composite Beads for Improved Performance of Recycle

Gao

et al. 2020

ACS Omega

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“…Similar low K m values were earlier reported for the peroxidase activity of Cyt c using graphene oxide, silica nanoparticles, and polymer scaffold as supporting systems. 32 , 38 , 39 The K m value of the Cyt c –MoS 2 nanobiohybrid is significantly lower than that of the free Cyt c (123 mM). 41 This suggests that Cyt c has a higher substrate binding affinity when adsorbed on the MoS 2 nanosheets.…”

Section: Resultsmentioning

confidence: 94%

Enhanced Catalytic Activity of a New Nanobiocatalytic System Formed by the Adsorption of Cytochrome c on Pluronic Triblock Copolymer Stabilized MoS₂ Nanosheets

et al. 2022

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The formation of nanobiohybrids through the immobilization of enzymes on functional nanomaterials has opened up exciting research opportunities at the nanobiointerfaces. These systems hold great promise for a wide range of applications in biosensing, biocatalytic, and biomedical fields. Here, we report the formation of a hybrid nanobiocatalytic system through the adsorption of cytochrome c (Cyt c ) on pluronic triblock copolymer, P123 (PEO- b -PPO- b -PEO), stabilized MoS 2 nanosheets. The use of pluronic polymer has helped not only to greatly stabilize the exfoliated MoS 2 nanosheets but also to allow easy adsorption of Cyt c on the nanosheets without major structural changes due to its excellent biocompatibility and soft protein-binding property. By comparing the catalytic activity of the Cyt c –MoS 2 nanobiohybrid with that of the free Cyt c and as-prepared MoS 2 nanosheets, we have demonstrated the active role of the nanobiointeractions in enhancing the catalytic activity of the hybrid. Slight structural perturbation at the active site of the Cyt c upon adsorption on MoS 2 has primarily facilitated the peroxidase activity of the Cyt c . As the MoS 2 nanosheets and the native Cyt c individually exhibit weaker intrinsic peroxidase activities, their mutual modulation at the nanobiointerface has made the Cyt c –MoS 2 a novel nanobiocatalyst with superior activity.

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“…157 The conjugation of poly(acrylic acid) (PAA) carboxyl groups to the primary amines in Cyt c using carbodiimide coupling (Figure 7a) resulted in a 34-fold increase in peroxidase turnover number (k cat ). 158 Cyt c−PAA conjugates were produced with molecular weight ranging from 1.8k to 250k g mol −1 , and it was noticed that the k cat value for guaiacol oxidation was increased with increasing M W of PAA (Figure 7b). Enzyme activity studies at pH 6−8 indicated improved activity for Cyt c−PAA conjugates at neutral or slightly alkaline pH.…”

Section: Lysine and The N-terminusmentioning

confidence: 99%

Recent Advances in Biocatalysis with Chemical Modification and Expanded Amino Acid Alphabet

et al. 2021

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The two main strategies for enzyme engineering, directed evolution and rational design, have found widespread applications in improving the intrinsic activities of proteins. Although numerous advances have been achieved using these ground-breaking methods, the limited chemical diversity of the biopolymers, restricted to the 20 canonical amino acids, hampers creation of novel enzymes that Nature has never made thus far. To address this, much research has been devoted to expanding the protein sequence space via chemical modifications and/or incorporation of noncanonical amino acids (ncAAs). This review provides a balanced discussion and critical evaluation of the applications, recent advances, and technical breakthroughs in biocatalysis for three approaches: (i) chemical modification of cAAs, (ii) incorporation of ncAAs, and (iii) chemical modification of incorporated ncAAs. Furthermore, the applications of these approaches and the result on the functional properties and mechanistic study of the enzymes are extensively reviewed. We also discuss the design of artificial enzymes and directed evolution strategies for enzymes with ncAAs incorporated. Finally, we discuss the current challenges and future perspectives for biocatalysis using the expanded amino acid alphabet.

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Cytochrome c–poly(acrylic acid) conjugates with improved peroxidase turnover number

Abstract: Cytochrome c–poly(acrylic acid) conjugates with 34-fold enhanced peroxidase activity due to acidification of enzyme microenvironment and suppression of wasteful intermediates.

Cited by 11 publications

References 33 publications

Immobilization of Pectinase onto Porous Hydroxyapatite/Calcium Alginate Composite Beads for Improved Performance of Recycle

Immobilization of Pectinase onto Porous Hydroxyapatite/Calcium Alginate Composite Beads for Improved Performance of Recycle

Enhanced Catalytic Activity of a New Nanobiocatalytic System Formed by the Adsorption of Cytochrome c on Pluronic Triblock Copolymer Stabilized MoS₂ Nanosheets

Recent Advances in Biocatalysis with Chemical Modification and Expanded Amino Acid Alphabet

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Cytochrome c–poly(acrylic acid) conjugates with improved peroxidase turnover number

Abstract: Cytochrome c–poly(acrylic acid) conjugates with 34-fold enhanced peroxidase activity due to acidification of enzyme microenvironment and suppression of wasteful intermediates.

Cited by 11 publications

References 33 publications

Immobilization of Pectinase onto Porous Hydroxyapatite/Calcium Alginate Composite Beads for Improved Performance of Recycle

Immobilization of Pectinase onto Porous Hydroxyapatite/Calcium Alginate Composite Beads for Improved Performance of Recycle

Enhanced Catalytic Activity of a New Nanobiocatalytic System Formed by the Adsorption of Cytochrome c on Pluronic Triblock Copolymer Stabilized MoS2 Nanosheets

Recent Advances in Biocatalysis with Chemical Modification and Expanded Amino Acid Alphabet

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Enhanced Catalytic Activity of a New Nanobiocatalytic System Formed by the Adsorption of Cytochrome c on Pluronic Triblock Copolymer Stabilized MoS₂ Nanosheets