title: Exploiting metagenomic diversity for novel polyhydroxyalkanoate synthases: production of a terpolymer poly(3-hydroxybutyrate-co-3-hydroxyhexanoate-co-3-hydroxyoctanoate) with a recombinant Pseudomonas putida strain.
abstract: A metagenomic library of 2.1×10(6) clones was constructed using oil-contaminated soil from Gujarat (India). One of the fosmid clones, 40N22, encodes a polyhydroxyalkanoate synthase showing 76% identity with an Alcaligenes sp. synthase. The corresponding gene was expressed in Pseudomonas putida KT2440 ΔphaC1 which is impaired in PHA production. The gene conferred the recombinant strain PpKT-40N22 with the ability to produce copolymers with up to 21% in medium-chain-length content. Thus, 37% and 45% of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) and poly(3-hydroxybutyrate-co-3-hydroxyhexanoate-co-3-hydroxyoctanoate), respectively were obtained when using sodium heptanoate and oleic acid as carbon sources. These 3-hydroxybutyrate-(3HB)-based polymers are of interest since they incorporate the properties of medium chain length polymers and thus increase the range of applications of PHAs.
authors.affiliation: Environmental Microbiology Laboratory, Helmholtz Centre for Infection Research, Inhoffenstrasse, 7, D-38124 Braunschweig, Germany.
authors.forename: Simrita
authors.initials: S
authors.lastname: Cheema
authors.forename: Monica
authors.initials: M
authors.lastname: Bassas-Galia
authors.forename: Priyangshu M
authors.initials: PM
authors.lastname: Sarma
authors.forename: Banwari
authors.initials: B
authors.lastname: Lal
authors.forename: Sagrario
authors.initials: S
authors.lastname: Arias
mesh_terms: D000217:Acyltransferases / Q000378:metabolism*; D001483:Base Sequence; D002244:Carbon / Q000494:pharmacology; D003470:Culture Media / Q000494:pharmacology; D005798:Genes, Bacterial / Q000235:genetics; D005820:Genetic Testing; D014644:Genetic Variation / Q000187:drug effects; D056186:Metagenomics / Q000379:methods*; D008969:Molecular Sequence Data; D010802:Phylogeny; D011091:Polyesters / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D016958:Pseudomonas putida / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics*; D011995:Recombination, Genetic / Q000235:genetics*; D017422:Sequence Analysis, DNA; D013379:Substrate Specificity / Q000187:drug effects; D000094122:Polyhydroxybutyrates
chemical_list: D000217:Acyltransferases; D002244:Carbon; D003470:Culture Media; D011091:Polyesters; C554196:poly(3-hydroxybutyrate-co-3-hydroxyhexanoate-co-3-hydroxyoctanoate); D000094122:Polyhydroxybutyrates; C077225:poly(3-hydroxyalkanoic acid) synthase