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Table 2 The KstD enzyme activity (U mg−1 total soluble protoplast protein and extracellular protein) of M. neoaurum DSM 1381, ΔkstD1, recombinant E. coli BL21 (DE3), and B. subtilis 6051a strains

From: Identification, function, and application of 3-ketosteroid Δ1-dehydrogenase isozymes in Mycobacterium neoaurum DSM 1381 for the production of steroidic synthons

Strains

Substrate

AD

4HP

Intracellular enzyme

Extracellular enzyme

Intracellular enzyme

Extracellular enzyme

DSM 1381

2.45 ± 0.05

u.d.

2.00 ± 0.19

u.d.

ΔkstD1

0.14 ± 0.01

u.d.

0.06 ± 0.01

u.d.

6051a-pHT01

u.d.

u.d.

u.d.

u.d.

6051a-kstD1

2.12 ± 0.06

u.d.

1.81 ± 0.05

u.d.

6051a-kstD2

22.40 ± 1.26

u.d.

19.19 ± 0.96

u.d.

6051a-kstD3

0.18 ± 0.06

u.d.

0.15 ± 0.06

u.d.

BL21-pET28a

u.d.

u.d.

u.d.

u.d.

BL21-kstD1

0.34 ± 0.04

u.d.

0.21 ± 0.05

u.d.

BL21-kstD2

3.46 ± 0.07

u.d.

2.82 ± 0.12

u.d.

BL21-kstD3

0.29 ± 0.08

u.d.

0.32 ± 0.14

u.d.

  1. All assays were performed with triplicate cultures. Standard deviations of the biological replicates are shown
  2. DSM1381, M. neoaurum DSM 1381; ΔkstD1, mutant of M. neoaurum DSM 1381 deleted kstD1; BL21-kstD1/BL21-kstD2/BL21-kstD3, E. coli BL21 (DE3) containing pET28a-kstD1/pET28a-kstD2/pET28a-kstD3; BL21-pET28a, E. coli BL21 (DE3) containing empty plasmid pET28a(+); 6051a-kstD1/6051a-kstD2/6051a-kstD3, B. subtilis 6051a containing pHT01-kstD1/pHT01-kstD2/pHT01-kstD3; 6051a-pHT01, B. subtilis 6051a containing empty plasmid pHT01
  3. u.d. undetectable enzyme activity