Characterization Of A Highly Active Polyhydroxyalkanoate Synthase

• Main Author: Chuah, Jo-Ann
• Format: Thesis
• Language: English
• Published: 2012
• Subjects: QH1 Natural history (General - Including nature conservation, geographical distribution)
• Online Access: http://eprints.usm.my/44792/1/CHUAH%20JO-ANN.pdf; http://eprints.usm.my/44792/
• Institution: Universiti Sains Malaysia

Polyhydroxyalkanoate (PHA) synthase from a locally isolated Chromobacterium sp. USM2 (PhaCCs) exhibited superior polymerizing ability and broad in vivo substrate specificity with preferences for short chain length (SCL) [3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV)] and medium chain length (MCL) [3-hydroxyhexanoate (3HHx)] monomers. For further characterization of the synthase, a Strep2-tagged PhaCCs for expression in and purification from Escherichia coli, was constructed in this study. In vitro enzymatic assay revealed an activity of 253 ± 13 U/mg for polymerization of 3-hydroxybutyryl-coenzyme A (3HB-CoA), which was approximately fivefold higher than that of model PHAproducing strain Cupriavidus necator (39 ± 5 U/mg).