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Rapid quantification of intracellular PHA using infrared spectroscopy: An application in mixed cultures

Paper ID Volume ID Publish Year Pages File Format Full-Text
23980 43486 2010 8 PDF Available
Title
Rapid quantification of intracellular PHA using infrared spectroscopy: An application in mixed cultures
Abstract

Fourier transform infrared (FT-IR) spectroscopy is proposed for a method for rapid quantification of polyhydroxyalkanoates (PHA) in mixed culture bacterial systems. Spectra from 122 samples from a wide range of PHA production systems were studied. The spectra were collected in a library that was used to calibrate a partial least squares (PLS) model linking FT-IR spectra with PHA content in the biomass.The library of spectra contained samples with a range of total PHA content (0.03–0.58 w/w) as well as varying compositions (poly-(3-hydroxyvalerate) (3HV) content of 0–63% in Cmol basis). A robust PLS model was developed using calibration data from a diverse range of systems and PHA content. Coupling this model with FT-IR spectra has been shown to be applicable for predicting PHA content in mixed culture production systems. The method was used to reliably determine PHA content in biomass from a new, independent PHA production system with a standard error of prediction (RMSEP) value of 0.023 w/w, despite the complexity of the matrices.This method reduces the analytical time for PHA quantification down to under 30 min (5 min handling time was achieved when FT-IR equipment was immediately available), and eliminates hazardous waste by-products. The work has demonstrated a level of accuracy and reproducibility in quantifying PHA in mixed culture systems similar to that obtained from the GC analytical technique. Further work is required to enable the use of the method to analyze crystallinity related factors that may be useful towards quantifying poly-(3-hydroxybutyrate) and poly-(3-hydroxyvalerate) (3HB/3HV) composition. The method has been shown to be suitable for rapid quantification in large scale applications and in its present form is reliable for routine process monitoring.

Keywords
Infrared spectroscopy; Polyhydroxyalkanoates; Mixed cultures
First Page Preview
Rapid quantification of intracellular PHA using infrared spectroscopy: An application in mixed cultures
Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 150, Issue 3, November 2010, Pages 372–379
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering