PHA Extraction, Quantitation and Calculation of Production Yield

PHAs were isolated as described in Snell et al. (2002) with some modification. Briefly 500 μL of culture were centrifuged (8000 rpm, 15 min) and stored at −20°C till analyze. PHAs were extracted and methanolysed by resuspending pellets in 500 μL of chloroform and 2 mL of methanolysis solution consisting in UHPLC methanol: concentrated HCl (90:10). The methanolysis solution also includes 0.1 mg/mL of 3-methylbenzoic acid as reaction house standard. Mixture was then incubated at 100°C during 2 h before being cooled down on ice. One milliliter of distilled water was then added and the bottom chloroform part was recovered and analyzed by GC-MS.