Characterization of 13 kDa granule-associated protein in Aeromonas caviae and biosynthesis of polyhydroxyalkanoates with altered molar composition by recombinant bacteria

Analysis of native poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) [P(3HB-co-3HHx)] inclusions from Aeromonas caviae FA440 revealed that ORF1 (a 348-bp gene located immediately upstream of phaC(Ac)) encodes a 13-kDa granule-associated protein, which was referred to as phaP(Ac). Several recombinant str...

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Bibliographic Details
Published in:Biomacromolecules 2001, Vol.2 (1), p.148-153
Main Authors: Fukui, T, Kichise, T, Iwata, T, Doi, Y
Format: Article
Language:English
Subjects:
Acyltransferases - metabolism
Aeromonas - enzymology
Aeromonas - genetics
Aeromonas - metabolism
Aeromonas caviae
Bacterial Proteins - chemistry
Bacterial Proteins - genetics
Bacterial Proteins - metabolism
Genes, Bacterial - genetics
Genetic Engineering
Inclusion Bodies - chemistry
Inclusion Bodies - enzymology
Molecular Weight
Olea
Polyesters - chemistry
Polyesters - metabolism
Pseudomonas putida - enzymology
Pseudomonas putida - genetics
Pseudomonas putida - metabolism
Substrate Specificity
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Summary:Analysis of native poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) [P(3HB-co-3HHx)] inclusions from Aeromonas caviae FA440 revealed that ORF1 (a 348-bp gene located immediately upstream of phaC(Ac)) encodes a 13-kDa granule-associated protein, which was referred to as phaP(Ac). Several recombinant strains of A. caviae were constructed and conducted to analyze their PHA-producing abilities. A transconjugant of FA440 harboring additional copies of phaPCJ(Ac) genes accumulated P(3HB-co-3HHx) copolyesters with much higher 3HHx composition (46-63 mol %) than wild-type strain from alkanoates or olive oil. Deletion analysis revealed that overexpression of phaJ(Ac) encoding monomer-supplying (R)-hydratase was not a reason for the compositional change in the recombinant strains. PHA synthase activity in PHA inclusion fraction from the transconjugant composed of 60 mol % of 3HHx was 10-fold higher than that from the strain FA440 with 13 mol % of 3HHx, suggesting an importance of the level of PHA synthase activity for controlling the PHA composition in vivo.
ISSN:1525-7797
DOI:10.1021/bm0056052