Intracellular depolymerase and polyhydroxyoctanoate granule integrity in Pseudomonas oleovorans.
When polyhydroxyoctanoate (PHO) was produced by Pseudomonas oleovorans during a regimen of intermittent feeding on octanoic acid, there was a significant change in both the polymer associated proteins and the composition of the enclosed polymer. The polymer granules were isolated with their protein coat intact and the enzymatic hydrolysis of the polymer within this cell free system was determined. The degradation rate for the PHO in these native granules reached a maximum of 1.17 mg/h at an optimum pH of 9 when incubated at 30 degrees C. A study of the effect of various inhibitors on depolymerase activity suggested that the enzyme most likely has disulfide linkages and serine residues at its active site. Ultrastructure studies suggested this loss of enzyme activity was correlated with significant organizational degeneration in the proteins associated with the PHO inclusion body. Once solubilized from the granule, the depolymerase itself remained enzymatically active, and addition of this released material to other granule preparations increased the rate of polymer granule degradation. Similarly, when colloidal suspensions of purified, amorphous PHO were placed in contact with that depolymerase, they also underwent rapid degradation. In contrast, when crystalline solvent-cast PHO films were placed in contact with this enzyme, no degradative activity was observed.
Department of Biochemistry and Molecular Biology, University of Massachusetts, Amherst 01003-4505, USA.
L J
LJ
Foster
E S
ES
Stuart
A
A
Tehrani
R W
RW
Lenz
R C
RC
Fuller
D001426:Bacterial Proteins / Q000378:metabolism; D002265:Carboxylic Ester Hydrolases / Q000737:chemistry / Q000378:metabolism*; D003594:Cytoplasmic Granules / Q000737:chemistry / Q000378:metabolism; D005285:Fermentation; D002479:Inclusion Bodies / Q000378:metabolism* / Q000648:ultrastructure*; D011091:Polyesters / Q000737:chemistry / Q000378:metabolism*; D011549:Pseudomonas / Q000254:growth & development / Q000378:metabolism*
D001426:Bacterial Proteins; D011091:Polyesters; C088151:poly(3-hydroxyoctanoic acid); D002265:Carboxylic Ester Hydrolases; C080782:poly(3-hydroxyoctanoic acid) depolymerase