Table 3 Kinetic parameters of GOase variants determined in the TiTR with HMF.

From: Toward scalable biocatalytic conversion of 5-hydroxymethylfurfural by galactose oxidase using coordinated reaction and enzyme engineering

Variant

KMO (mM)

KMS (mM)

kcat (s−1)

kcat/KMO (M−1 s−1)

kcat/KMS (M−1 s−1)

M1

0.16 ± 0.11

53 ± 18

23.5 ± 4.7

(1.5 ± 1.1) × 105

(0.04 ± 0.02) × 104

M3–5a

1.39 ± 0.46

14.9 ± 4.5

651 ± 132

(4.7 ± 1.8) × 105

(4.4 ± 1.6) × 104

M4

0.37 ± 0.07

1.83 ± 0.30

120 ± 6

(3.3 ± 0.6) × 105

(6.6 ± 1.1) × 104

M6-A

0.15 ± 0.02

6.19 ± 0.41

166 ± 4

(10.9 ± 1.5) × 105

(2.7 ± 0.2) × 104

  1. KMO: KM for oxygen, KMS: KM for HMF. aMeasured using CFE powder with an estimated GOase content, giving a higher uncertainty in the kcat parameter. Data previously published in the PhD thesis of A. T. P.77.
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