Metabolism of poly- -hydroxybutyrate: effect of mild alkaline extraction on native poly- -hydroxybutyrate granules.
Mild alkaline extraction of native poly-beta-hydroxybutyrate (PHB) granules results in the solubilization of a protein fraction. Both the solubilized protein fraction and the extracted granules are essentially devoid of PHB synthetase activity unless recombined. The protein fraction has been separated by chromatography into two components (A-I and A-II). A-I but not A-II can be recombined with extracted granules to give rise to PHB synthetase activity. Extracted granules no longer require pretreatment with activator or trypsin but are directly susceptible to hydrolysis by Rhodospirillum rubrum depolymerase. Addition of A-II or A-I prevents the direct hydrolysis by depolymerase. The inhibition is reversed by activator or trypsin. We conclude that native granules are associated with a protein inhibitor which prevents the hydrolysis of PHB by depolymerase unless the protein is destroyed by trypsin, removed by alkaline extraction, or modified by activator.
D001410:Bacillus megaterium; D001426:Bacterial Proteins / Q000302:isolation & purification / Q000378:metabolism; D002247:Carbon Isotopes; D002474:Cell-Free System; D002854:Chromatography, Paper; D004589:Electrophoresis, Disc; D006868:Hydrolysis; D006885:Hydroxybutyrates / Q000096:biosynthesis / Q000378:metabolism*; D008025:Ligases / Q000378:metabolism; D006899:Mixed Function Oxygenases / Q000378:metabolism; D008970:Molecular Weight; D011108:Polymers / Q000378:metabolism*; D011549:Pseudomonas / Q000201:enzymology; D012246:Rhodospirillum / Q000201:enzymology; D012972:Sodium Hydroxide; D012997:Solvents; D014357:Trypsin
D001426:Bacterial Proteins; D002247:Carbon Isotopes; D006885:Hydroxybutyrates; D011108:Polymers; D012997:Solvents; D012972:Sodium Hydroxide; D006899:Mixed Function Oxygenases; D014357:Trypsin; D008025:Ligases