Techniques for tracing PHA‐producing organisms and for qualitative and quantitative analysis of intra‐ and extracellular PHA

Abstract

Poly(hydroxyalkanoates) (PHAs) constitute a versatile family of prokaryotic reserve materials that, thanks to their plastic‐like properties, displays high potential for application in various fields of the plastic market. Various methods are reported for tracing PHA‐producing organisms, and for qualitative and quantitative analysis of both intra‐ and extracellular PHA. Different techniques are needed first to discover novel powerful PHA‐producing microbes, second to identify new PHA‐building blocks, and thirdly for structural and conformational PHA analysis on the intramolecular level. Further, as the aspect of utmost industrial significance, methods for fast, reliable, and routine process control during PHA bioproduction are required. This review tracks the development of different methods for PHA detection and determination, such as gravimetry, turbidimetry, optical, fluorescence, or electron microscopic techniques, UV, Raman, and infrared spectrometry, genetic methods (Southern blot hybridization, PCR, fluorescence‐labeled in situ hybridization), enzyme‐based biosensors, 1H and 13C NMR, and advanced gas and liquid chromatographic approaches. We propose the most suitable techniques for particular challenges of PHA research and production, and critically assess their strengths and limitations. Moreover, we provide suggestions for future developments in order to accelerate the final launch of PHA on the “bioplastic” market as anticipated since decades by the respective scientific community.

Production of Polyhydroxyalkanoate (PHA) Biopolyesters by Extremophiles?

“…Tracing halophile PHA producers: Numerous techniques for tracing PHA in various different microbes were described in the past [82]. In the case of halophiles, existing PHA detection methods are not precise enough, frequently deliver false positive results, and/or need prior PHA synthesis before the screening.…”

Microbial synthesis of a novel terpolyester P(LA‐co‐3HB‐co‐3HP) from low‐cost substrates

“…Polyhydroxyalkanoates (PHA) are a family of diverse polyesters synthesized by a variety of bacteria as intracellular carbon and energy storage compounds (Li et al, 2007;Chen and Hajnal, 2015;Koller and Rodr ıguez-Contreras, 2015). As biorenewable and biodegradable materials, the diversity of PHA provides different physical properties to suit various applications (Chen, 2009;Brigham and Sinskey, 2012;Koller, 2014).…”

Quantitative Raman Spectroscopy Analysis of Polyhydroxyalkanoates Produced by Cupriavidus necator H16

“…The most commonly employed method for quantitative analysis of PHAs is gas chromatography; nevertheless, the labor intensity and time demands of this method prevents its application for routine analysis of PHAs during the biotechnological process. Apart from gas chromatography, there are other techniques (such as gravimetry, turbidimetry, UV spectroscopy, optical fluorescence, and electron microscopy) that can be used for the determination of PHAs in bacterial cells [4], but most of them are time consuming or do not provide sufficient reliability or sensitivity to be used in the PHA production process control.…”