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Table 2 Calculated apparent kinetic parameters of each mutant compared to wt NicA2

From: Optimization of a nicotine degrading enzyme for potential use in treatment of nicotine addiction

Variant

kcat/Km (M−1 s−1)

(apparent)

kcat (s−1)

(apparent)

Km (nM)

(apparent)

wt NicA2

6.4 × 104

0.015

240

A107R

1.9 × 105

0.159

830

A107T

1.8 × 105

0.050

285

A107G

1.6 × 105

0.053

335

F104 T

1.7 × 105

0.055

330

F104I

1.6 × 105

0.039

240

F104R

1.5 × 105

0.041

270

F104S

1.3 × 105

0.031

250

G106A

1.1 × 105

0.031

280

F355H

1.3 × 105

0.029

220

A426C

1.3 × 105

0.030

230

  1. Apparent kcat/Km values were calculated using the low nicotine assay results (Fig. 2c) using a form of the Michaelis-Menten equation (v0/[E] = kcat/Km*[S]) and to construct progress curves that were fitted to a simple kinetic model (KinTek) to derive apparent kcat values. The resulting kcat/Km and kcat values were used to calculate the apparent Km values
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BMC Biotechnology

ISSN: 1472-6750

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