Kinetic evidence for surface residues influencing the active site of Coprinus cinereus peroxidase: analysis of the pH dependence of G154E, P90H and P90H–G154E substrate entrance mutants

Cerbo, Paola Di; Welinder, Karen G.; Schiødt, Christine Bruun

doi:10.1016/s0167-4838(00)00201-6

Cited by 3 publications

(7 citation statements)

References 24 publications

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“…The unfolding experiments (Table 4) showed that such a stabilizing salt bridge was not present in the similar double mutant of CIP. A similar conclusion was recently reached from enzyme kinetic studies of compound I formation of the single and double mutants (20).…”

Section: Discussionsupporting

confidence: 82%

Kinetic Stability of Designed Glycosylation Mutants of Coprinus cinereus Peroxidase

Tams

Welinder

2001

Biochemical and Biophysical Research Communications

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Section: Discussionsupporting

confidence: 82%

Kinetic Stability of Designed Glycosylation Mutants of Coprinus cinereus Peroxidase

Tams

Welinder

2001

Biochemical and Biophysical Research Communications

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“…To reveal the mechanism of how to control the compound I formation rate, we examined the correlation between the push and pull effects and the compound I formation rate. The average compound I formation rates, calculated using data from at least four studies, were utilized (see Table ) − . Because the availability of kinetic data for ARP/CIP at a pH value of 7.0−8.0 is limited, the compound I formation rate for ARP was determined using the transient-state kinetic technique.…”

Section: Resultsmentioning

confidence: 99%

“…While peroxidases have common structural characteristics to react with hydrogen peroxide as mentioned above, rates of compound I formation among peroxidases are significantly different. CcP and APX, which belong to class I peroxidase, form compound I with 10 7 −10 8 M −1 s −1 whereas LiP, MnP, and ARP, which are classified as a class II peroxidase, show ∼100-fold slower formation rates, 10 5 −10 6 M −1 s −1 , and HRP, a class III peroxidase, has ∼10-fold slower rate − . These results suggest the strength of the push and pull effects is modulated in each class of peroxidases.…”

mentioning

confidence: 93%

Paramagnetic ¹³C and ¹⁵N NMR Analyses of Cyanide- (¹³C¹⁵N-) Ligated Ferric Peroxidases: The Push Effect, Not Pull Effect, Modulates the Compound I Formation Rate

2009

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Paramagnetic (13)C and (15)N NMR spectroscopy of heme-bound cyanide ((13)C(15)N) was utilized to quantitatively distinguish the electron donor effect (the push effect) from the proximal histidine and hydrogen-bonding effect (the pull effect) from the distal amino acid residues in cytochrome c peroxidase (CcP), ascorbate peroxidase (APX), lignin peroxidase (LiP), and manganese peroxidase (MnP). Paramagnetic (13)C NMR signals of heme-bound (13)C(15)N of these peroxidases were observed in a wide range, -3501 ppm (CcP), -3563 ppm (APX), -3823 ppm (MnP), and -3826 ppm (LiP), while paramagnetic (15)N NMR signals of those were detected in a narrow range, 574 ppm (ARP), 605 ppm (CcP), 626 ppm (LiP), and 654 ppm (MnP). Detailed analysis, combined with the previous results for horseradish peroxidase and Arthromyces ramosus peroxidase, indicated that the push effect is quite different among these peroxidases while the pull effect is similar. More importantly, a strong correlation between the (13)C NMR shift (the push effect) and the compound I formation rate was observed, indicating that the push effect causes a variation in the compound I formation rate. Comparison of the (13)C and (15)N NMR results of these peroxidases with their crystal structures suggests that the orientation of the proximal imidazole plane to the heme N-Fe-N axis controls the push effect and the compound I formation rate of peroxidase.

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“…Peroxidases (EC 1.11.1.7) are heme-containing enzymes that are widely distributed in animals, plants and microorganisms. They oxidize a variety of organic and inorganic compounds with the presence of hydrogen peroxide 1,2,3,4 . Their powerful oxidizing ability makes peroxidases very useful in many fields, including analytical chemistry, immunochemistry, and biosensor construction.…”

Section: Introductionmentioning

confidence: 99%

“…Their powerful oxidizing ability makes peroxidases very useful in many fields, including analytical chemistry, immunochemistry, and biosensor construction. In addition, they have great potential and prospect in the textile, paper and pulp bleaching industries 1,5 . Although peroxidases are important in many industries, it is difficult to produce these enzymes in large scale, because most of the peroxidases are isolated from plants.…”

Section: Introductionmentioning

confidence: 99%

Optimization of an Activity Assay of Coprinus Cinereus Peroxidase

Dong¹,

Zhang²,

et al. 2018

Braz. arch. biol. technol.

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To seek a simple, rapid and sensitive Coprinus cinereus Peroxidase (CIP) activity assay, a convenient one-factor-ata-time (OFAT) method and a response surface methodology (RSM) were used. The recombinant CIP expressed in Pichia pastoris was purified with the Ni-NTA spin column. Based on the results of catalytic efficiency (kcat/Km) analysis, 2,2'-azinobis (ethylbenzthiazoline-6-sulfonate) (ABTS) was selected as the optimal enzyme substrate. Results of the OFAT method showed that enzymatic reaction performed in 0.1 mol/L sodium acetate (pH 5.0) buffer in a 200-µl reaction mixture containing 0.5 mmol/L ABTS, 10 mmol/L hydrogen peroxide (H2O2), 49.7 ng CIP at 25°C gave an average CIP activity of 88 U/mL. The ABTS and H2O2 concentrations were then further optimized to improve the sensitivity of the assay. To do that, RSM was conducted through central composite design, and a reduced quadratic model with good fit regression equation was generated. ANOVA analysis of this model indicated that the concentrations of ABTS and H2O2 and their interaction had significant impact on the assay sensitivity. The optimal reaction mixture was determined to include an initial ABTS concentration of 0.82 mmol/L 49.7 ng CIP and 16.36 mmol/L H2O2, and the activity under this condition was determined to be 138.89 U/mL.

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Kinetic evidence for surface residues influencing the active site of Coprinus cinereus peroxidase: analysis of the pH dependence of G154E, P90H and P90H–G154E substrate entrance mutants

Cited by 3 publications

References 24 publications

Kinetic Stability of Designed Glycosylation Mutants of Coprinus cinereus Peroxidase

Kinetic Stability of Designed Glycosylation Mutants of Coprinus cinereus Peroxidase

Paramagnetic ¹³C and ¹⁵N NMR Analyses of Cyanide- (¹³C¹⁵N-) Ligated Ferric Peroxidases: The Push Effect, Not Pull Effect, Modulates the Compound I Formation Rate

Optimization of an Activity Assay of Coprinus Cinereus Peroxidase

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Kinetic evidence for surface residues influencing the active site of Coprinus cinereus peroxidase: analysis of the pH dependence of G154E, P90H and P90H–G154E substrate entrance mutants

Cited by 3 publications

References 24 publications

Kinetic Stability of Designed Glycosylation Mutants of Coprinus cinereus Peroxidase

Kinetic Stability of Designed Glycosylation Mutants of Coprinus cinereus Peroxidase

Paramagnetic 13C and 15N NMR Analyses of Cyanide- (13C15N-) Ligated Ferric Peroxidases: The Push Effect, Not Pull Effect, Modulates the Compound I Formation Rate

Optimization of an Activity Assay of Coprinus Cinereus Peroxidase

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Paramagnetic ¹³C and ¹⁵N NMR Analyses of Cyanide- (¹³C¹⁵N-) Ligated Ferric Peroxidases: The Push Effect, Not Pull Effect, Modulates the Compound I Formation Rate