TY - JOUR
T1 - Fast method for the determination of short-chain-length polyhydroxyalkanoates (scl-PHAs) in bacterial samples by In Vial-Thermolysis (IVT)
AU - Abbondanzi, F.
AU - Biscaro, G.
AU - Carvalho, G.
AU - Favaro, L.
AU - Lemos, P.
AU - Paglione, M.
AU - Samorì, C.
AU - Torri, C.
N1 - Sem pdf conforme despacho.
University of Padova through the Progetto di Ateneo - CPDA137517/13 ;
FCT/MEC (PEst-C/EQB/LA0006/2013)
PY - 2017/10/25
Y1 - 2017/10/25
N2 - A new method based on the GC–MS analysis of thermolysis products obtained by treating bacterial samples at a high temperature (above 270 °C) has been developed. This method, here named “In-Vial-Thermolysis” (IVT), allowed for the simultaneous determination of short-chain-length polyhydroxyalkanoates (scl-PHA) content and composition. The method was applied to both single strains and microbial mixed cultures (MMC) fed with different carbon sources. The IVT procedure provided similar analytical performances compared to previous Py-GC–MS and Py-GC-FID methods, suggesting a similar application for PHA quantitation in bacterial cells. Results from the IVT procedure and the traditional methanolysis method were compared; the correlation between the two datasets was fit for the purpose, giving a R2 of 0.975. In search of further simplification, the rationale of IVT was exploited for the development of a “field method” based on the titration of thermolyzed samples with sodium hydrogen carbonate to quantify PHA inside bacterial cells. The accuracy of the IVT method was fit for the purpose. These results lead to the possibility for the on-line measurement of PHA productivity. Moreover, they allow for the fast and inexpensive quantification/characterization of PHA for biotechnological process control, as well as investigation over various bacterial communities and/or feeding strategies.
AB - A new method based on the GC–MS analysis of thermolysis products obtained by treating bacterial samples at a high temperature (above 270 °C) has been developed. This method, here named “In-Vial-Thermolysis” (IVT), allowed for the simultaneous determination of short-chain-length polyhydroxyalkanoates (scl-PHA) content and composition. The method was applied to both single strains and microbial mixed cultures (MMC) fed with different carbon sources. The IVT procedure provided similar analytical performances compared to previous Py-GC–MS and Py-GC-FID methods, suggesting a similar application for PHA quantitation in bacterial cells. Results from the IVT procedure and the traditional methanolysis method were compared; the correlation between the two datasets was fit for the purpose, giving a R2 of 0.975. In search of further simplification, the rationale of IVT was exploited for the development of a “field method” based on the titration of thermolyzed samples with sodium hydrogen carbonate to quantify PHA inside bacterial cells. The accuracy of the IVT method was fit for the purpose. These results lead to the possibility for the on-line measurement of PHA productivity. Moreover, they allow for the fast and inexpensive quantification/characterization of PHA for biotechnological process control, as well as investigation over various bacterial communities and/or feeding strategies.
KW - Crotonic acid
KW - Mixed microbial cultures (MMC)
KW - Polyhydroxyalkanoates
KW - Thermolysis-GC–MS
KW - Thermolysis-titration
UR - http://www.scopus.com/inward/record.url?scp=85020430674&partnerID=8YFLogxK
U2 - 10.1016/j.nbt.2017.05.012
DO - 10.1016/j.nbt.2017.05.012
M3 - Article
C2 - 28591645
AN - SCOPUS:85020430674
SN - 1871-6784
VL - 39
SP - 29
EP - 35
JO - New Biotechnology
JF - New Biotechnology
ER -