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Table 2 Apparent kinetic parameters of native and recombinant sterol esterases from O. piceae on p -nitrophenol, glycerol and cholesterol esters

From: Recombinant sterol esterase from Ophiostoma piceae: an improved biocatalyst expressed in Pichia pastoris

Substrate

Acyl length and insaturation(s)

Catalytic

Native

Recombinant

  

parameters

OPEa

OPE

p -Nitrophenyl butyrate

C4:0

K m app

0.27±0.03

2.33±0.19

k cat app

44±2

2533±55

  

kcatapp/Kmapp

162±13

1089±70

p -Nitrophenyl palmitate

C16:0

K m app

0.33±0.03

0.37±0.03

k cat app

74±3

1049±28

kcatapp/Kmapp

224±12

2875±212

Glyceryl tributyrate

C4:0

K m app

9.90±0.80

5.10±0.30

k cat app

179±4

1041±14

  

kcatapp/Kmapp

18±1

204±10

Glyceryl trioleate

C18:1

K m app

0.98±0.08

0.71±0.09

k cat app

290±7

1362±41

kcatapp/Kmapp

296±18

1924±197

Cholesteryl butyrate

C4:0

K m app

3.00±0.50

1.60±0.20

k cat app

47±2

212±7

kcatapp/Kmapp

15.6±1.8

133±13

Cholesteryl oleate

C18:1

K m app

1.00±0.10

0.69±0.10

k cat app

138±4

631±21

kcatapp/Kmapp

138±9

918±108

Cholesteryl linoleate

C18:2

K m app

0.99±0.06

0.71±0.09

  

k cat app

150±3

798±25

  

kcatapp/Kmapp

152±6

1132±123

  1. Reactions were carried out in the presence of the non-ionic detergent Genapol X-100.
  2. Kmapp (mM), kcatapp (s-1) and kcatapp/Kmapp (s-1mM-1). Standard errors are based on the curve fitting using Sigma plot 11.0 software. The kcatapp/Kmapp standard errors were obtained by fitting the normalized Michaelis-Menten equation as V = (kcatapp/Kmapp)[S]/(1 + [S]/Kmapp). Kinetics parameters were obtained using the average molecular weight of the non-deglycosylated recombinant protein obtained by mass spectrometry as was previously considered with native one.
  3. a[32].