Properties and biodegradability of ultra-high-molecular-weight poly[(R)-hydroxybutyrate] produced by a recombinant Escherichia coli.
Ultra-high-molecular-weight poly[(R)-3-hydroxybutyrate] (P(3HB)) (Mw = 3-11 x 10(6)) was produced from glucose by a recombinant Escherichia coli XL1-Blue (pSYL105) harboring Ralstonia eutropha H16 polyhydroxyalkanoate (PHA) biosynthesis genes. Morphology of ultra-high-molecular-weight P(3HB) granules in the recombinant cells was studied by transmission electron microscopy. The recombinant E. coli contained several P(3HB) granules within a cell. Freeze-fracture morphology of ultra-high-molecular-weight P(3HB) granules showed the needle-type as that of P(3HB) granules in R. eutropha. Both the P(3HB) granules in wet cells and wet native granules isolated from the recombinant cells proved to be amorphous on the X-ray diffraction patterns. Mechanical properties of ultra-high-molecular-weight P(3HB) films were markedly improved by stretching over 400%, resulting from high crystallinity and highly oriented crystal regions. Biodegradability of the films of ultra-high-molecular-weight P(3HB) was tested with an extracellular polyhydroxybutyrate depolymerase from Alcaligenes faecalis T1. The rate of enzymatic erosion of P(3HB) films was not dependent of the molecular weight but was dependent of the crystallinity. In addition, it is demonstrated that all ultra-high-molecular-weight P(3HB) films were completely degraded at 25 degrees C in a natural river freshwater within 3 weeks.
Kusaka
S
S
Akebono Brake R&D Centre, Ltd., Saitama, Japan.
Iwata
T
T
Doi
Y
Y
D000217:Acyltransferases / Q000235:genetics / Q000378:metabolism*; D000421:Alcaligenes / Q000201:enzymology* / Q000235:genetics; D001673:Biodegradation, Environmental; D003001:Cloning, Molecular / Q000379:methods; D003594:Cytoplasmic Granules / Q000378:metabolism / Q000648:ultrastructure; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism* / Q000648:ultrastructure; D006885:Hydroxybutyrates / Q000737:chemistry* / Q000378:metabolism* / Q000493:pharmacokinetics; D008970:Molecular Weight; D011091:Polyesters / Q000737:chemistry* / Q000378:metabolism* / Q000493:pharmacokinetics; D011994:Recombinant Proteins / Q000378:metabolism; D011995:Recombination, Genetic; D000094122:Polyhydroxybutyrates
D000217:Acyltransferases; D006885:Hydroxybutyrates; D011091:Polyesters; D011994:Recombinant Proteins; C003182:poly-beta-hydroxybutyrate; D000094122:Polyhydroxybutyrates; C077225:poly(3-hydroxyalkanoic acid) synthase