Rapid and direct compositional analysis of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) in whole bacterial cells by thermally assisted hydrolysis and methylation-gas chromatography.
Thermally assisted hydrolysis and methylation-gas chromatography (THM-GC) in the presence of an organic alkali was applied to the direct analysis of copolymer composition for poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [P(3HB-co-3HV)] accumulated in whole bacterial cells. Cupriavidus necator was grown on a liquid medium with different molar ratios of valeric acid (V) to glucose (G) in order to control the compositions of P(3HB-co-3HV) produced in the cells. Trace amounts (0.03 mg) of dried Cupriavidus necator cells were directly subjected to THM-GC in the presence of tetramethylammonium hydroxide (TMAH) at 400°C. The obtained chromatograms clearly showed nine characteristic peaks, attributed to the THM products from 3HB and 3HV units in the polymer chains, without any appreciable interference by the bacterial matrix components. Based on these peak intensities, the copolymer compositions were determined rapidly without using any cumbersome sample pretreatment. Moreover, the compositions thus obtained were in good agreement with those obtained by the conventional technique.
Department of Biological Chemistry, College of Bioscience and Biotechnology, Chubu University.
Siti
S
Baidurah
Yasuko
Y
Kubo
Mitsuhiro
M
Kuno
Kazuho
K
Kodera
Yasuyuki
Y
Ishida
Tsuneo
T
Yamane
Hajime
H
Ohtani
D020155:3-Hydroxybutyric Acid / Q000737:chemistry*; D002849:Chromatography, Gas / Q000379:methods*; D020118:Cupriavidus necator / Q000166:cytology*; D006868:Hydrolysis; D008745:Methylation; D010421:Pentanoic Acids / Q000737:chemistry*; D011108:Polymers / Q000737:chemistry*; D013696:Temperature; D013997:Time Factors
D010421:Pentanoic Acids; D011108:Polymers; C013056:beta-hydroxyvaleric acid; D020155:3-Hydroxybutyric Acid