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Table 2 In vivo and in vitro characterization of selected HisA/PriA homologs and mutants

From: Insights into the evolution of enzyme substrate promiscuity after the discovery of (βα)8 isomerase evolutionary intermediates from a diverse metagenome

Enzymes

In vivo activity

In vitro activitya

ProFAR isomerase (HisA)

PRA isomerase (TrpF)

HisA

TrpF

K M μM

k cat s−1

k cat/K M s−1 μM−1

K M μM

k cat s−1

k cat /K M s−1 μM−1

HisA_Afer

+

-

1.1 ± 0.2

0.05 ± 0.001

0.045

n.d.

n.d.

n.d.

PriA_Blon

+

+

2.7 ± 0.5

0.4 ± 0.1

0.1

6.1 ± 0.1

2.1 ± 0.5

0.3

CAM1

+

+

1.7 ± 0.1

0.3 ± 0.03

0.2

40 ± 7

3.5 ± 0.04

0.09

CAM1_A81G

+

+

1.7 ± 0.2

0.1 ± 0.01

0.06

32.2 ± 1.7

1.9 ± 0.1

0.06

CAM1_A81S

+

+

4.0 ± 0.9

0.2 ± 0.03

0.04

23.5 ± 6.5

0.5 ± 0.1

0.02

CAM2

+

-

n.d.

n.d.

n.d.

n.d.

n.d.

n.d.

  1. aEach data point comes from at least three independent determinations using freshly purified enzyme. n.d., activity not detected