In vivo deuteration of a native bacterial biopolymer for structural elucidation using SANS

• Published in: Physica. B, Condensed matter Vol. 350; no. 1; pp. E643 - E646
• Main Authors: Holden, P.J, Russell, R.A, Stone, D.J.M, Garvey, C.J, Foster, L.J.R
• Format: Journal Article
• Language: English
• Published: Elsevier B.V 15-07-2004
• Subjects: Biodeuteration; Polyhydroxyalkanoate; SANS; 27.10; Biodeuteration; SANS; 87.15; 61.12.E; Polyhydroxyalkanoate
• ISSN: 0921-4526
• DOI: 10.1016/j.physb.2004.03.172

In order to facilitate future structural studies, biodeuteration of bacterial polyhydroxyalkanoates (PHAs) was investigated. We report here the in vivo deuteration of poly 3-hydroxyoctanoate (PHO) produced by its native host, the bacterium
Pseudomonas oleovorans. Bacterial biomass was produced in bioreactor studies by growth on hydrogenated substrates and PHO was subsequently produced intracellularly (10–20% w/w) during batch fed growth on deuterated octanoic acid under oxygen limitation. GC-MS analyses of the PHO demonstrated that 13 of the 15 hydrogen atoms had been replaced with deuterium (except in position 3), the remaining two hydrogen presumably being derived from water. A SANS contrast variation study was conducted on whole cells and the results indicate the potential to discriminate inclusion bodies formed from deuterated precursor from an otherwise hydrogenated background.