Abstract
The activity of the proline catabolic enzyme pyrroline-5-carboxylate dehydrogenase (EC 1.5.1.12) was induced up to threehundredfold by the addition of three hundred proline to the growth medium of the Gram-positive bacterium Streptomyces coelicolor A3(2). Rifampicin, an inhibitor of RNA polymerase activity, abolished induction, implying that regulation was at the level of activation of gene transcription. The enzyme was purified and SDS-PAGE of the highly purified enzyme preparation revealed a single subunit with M(r) 68 000. A single band of protein, which also stained for enzyme activity, was observed after native gel electrophoresis. The M(r) of the enzyme was estimated to be approximately 265 000 by native gel electrophoresis and approximately 305 000 by gel filtration, which indicated that the enzyme had a tetrameric quaternary structure. The apparent K(m) for pyrroline-5-carboxylate was 109 ± 7.3 μM, whilst that for NAD+ was 43.3 ± 2.5 μM. Product inhibition by NADH (apparent K(i) 0.6 mM) was observed. The observed V(max) was 22.0 ± 1 mol min-1 (mg protein)-1. Neither 1 nor 5 mM proline had any effect on enzyme activity,, whilst glutamate was a very weak inhibitor.
| Original language | English |
|---|---|
| Pages (from-to) | 1739-1744 |
| Number of pages | 6 |
| Journal | Microbiology |
| Volume | 141 |
| Issue number | 7 |
| Publication status | Published - Jan 1 1995 |
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All Science Journal Classification (ASJC) codes
- Microbiology
Cite this
Interaction between primary and secondary metabolism in Streptomyces coelicolor A3(2) : Role of pyrroline-5-carboxylate dehydrogenase. / Drummond, Douglas Robert; Smith, S.; Wood, N. J.; Hodgson, D. A.
In: Microbiology, Vol. 141, No. 7, 01.01.1995, p. 1739-1744.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Interaction between primary and secondary metabolism in Streptomyces coelicolor A3(2)
T2 - Role of pyrroline-5-carboxylate dehydrogenase
AU - Drummond, Douglas Robert
AU - Smith, S.
AU - Wood, N. J.
AU - Hodgson, D. A.
PY - 1995/1/1
Y1 - 1995/1/1
N2 - The activity of the proline catabolic enzyme pyrroline-5-carboxylate dehydrogenase (EC 1.5.1.12) was induced up to threehundredfold by the addition of three hundred proline to the growth medium of the Gram-positive bacterium Streptomyces coelicolor A3(2). Rifampicin, an inhibitor of RNA polymerase activity, abolished induction, implying that regulation was at the level of activation of gene transcription. The enzyme was purified and SDS-PAGE of the highly purified enzyme preparation revealed a single subunit with M(r) 68 000. A single band of protein, which also stained for enzyme activity, was observed after native gel electrophoresis. The M(r) of the enzyme was estimated to be approximately 265 000 by native gel electrophoresis and approximately 305 000 by gel filtration, which indicated that the enzyme had a tetrameric quaternary structure. The apparent K(m) for pyrroline-5-carboxylate was 109 ± 7.3 μM, whilst that for NAD+ was 43.3 ± 2.5 μM. Product inhibition by NADH (apparent K(i) 0.6 mM) was observed. The observed V(max) was 22.0 ± 1 mol min-1 (mg protein)-1. Neither 1 nor 5 mM proline had any effect on enzyme activity,, whilst glutamate was a very weak inhibitor.
AB - The activity of the proline catabolic enzyme pyrroline-5-carboxylate dehydrogenase (EC 1.5.1.12) was induced up to threehundredfold by the addition of three hundred proline to the growth medium of the Gram-positive bacterium Streptomyces coelicolor A3(2). Rifampicin, an inhibitor of RNA polymerase activity, abolished induction, implying that regulation was at the level of activation of gene transcription. The enzyme was purified and SDS-PAGE of the highly purified enzyme preparation revealed a single subunit with M(r) 68 000. A single band of protein, which also stained for enzyme activity, was observed after native gel electrophoresis. The M(r) of the enzyme was estimated to be approximately 265 000 by native gel electrophoresis and approximately 305 000 by gel filtration, which indicated that the enzyme had a tetrameric quaternary structure. The apparent K(m) for pyrroline-5-carboxylate was 109 ± 7.3 μM, whilst that for NAD+ was 43.3 ± 2.5 μM. Product inhibition by NADH (apparent K(i) 0.6 mM) was observed. The observed V(max) was 22.0 ± 1 mol min-1 (mg protein)-1. Neither 1 nor 5 mM proline had any effect on enzyme activity,, whilst glutamate was a very weak inhibitor.
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UR - http://www.scopus.com/inward/citedby.url?scp=0029094244&partnerID=8YFLogxK
M3 - Article
C2 - 7551040
AN - SCOPUS:0029094244
VL - 141
SP - 1739
EP - 1744
JO - Microbiology
JF - Microbiology
SN - 1350-0872
IS - 7
ER -