Detailed algal extracellular carbohydrate-protein characterisation lends insight into algal solid-liquid separation process outcomes

N R H Rao; A M Granville; P R Wich; R K Henderson

doi:10.1016/j.watres.2020.115833

Detailed algal extracellular carbohydrate-protein characterisation lends insight into algal solid-liquid separation process outcomes

Water Res. 2020 Jul 1:178:115833. doi: 10.1016/j.watres.2020.115833. Epub 2020 Apr 21.

Authors

N R H Rao¹, A M Granville², P R Wich², R K Henderson³

Affiliations

¹ Algae and Organic Matter Laboratory (AOM Lab), School of Chemical Engineering, The University of New South Wales, Sydney, NSW, 2052, Australia; Centre for Advanced Macromolecular Design (CAMD), School of Chemical Engineering, The University of New South Wales, Sydney, NSW, 2052, Australia.
² Centre for Advanced Macromolecular Design (CAMD), School of Chemical Engineering, The University of New South Wales, Sydney, NSW, 2052, Australia.
³ Algae and Organic Matter Laboratory (AOM Lab), School of Chemical Engineering, The University of New South Wales, Sydney, NSW, 2052, Australia. Electronic address: r.henderson@unsw.edu.au.

PMID: 32339864
DOI: 10.1016/j.watres.2020.115833

Abstract

The effectiveness of algal solid-liquid separation processes has been impacted by the strong influence of algal extracellular organic matter (EOM), where the composition of proteins and carbohydrates and their associated interactions have been implicated. However, despite this, no studies have analysed the detailed protein and carbohydrate composition in EOM in relation to their impacts on separation. Hence, the aim of this study was to explore the relationship between the variety of carbohydrates and proteins present in the EOM of select algal and cyanobacterial samples and the associated separation performance to better understand the influence of specific biopolymers. The protein and carbohydrate composition of the EOM of three species - Microcystis aeruginosa CS-555/1, Chlorella vulgaris CS-42/7 and Microcystis aeruginosa CS-564/01, previously observed to result in variable treatment performance were investigated. The carbohydrates were analysed via high-performance anion-exchange chromatography (HPAEC) with pulsed amperometric detection (PAD) while the proteins were analysed using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) combined with liquid chromatography-mass spectrometry (LC-MS). Ten unique monosaccharides were identified; of these, the greatest proportion of charged uronic acid carbohydrates were present in the EOM of M. aeruginosa CS-564/01. The protein profiling revealed that M. aeruginosa CS-564/01 had a greater proportion and concentration of proteins >75 kDa when compared to M. aeruginosa CS-555/1 or C. vulgaris CS-42/7. It was determined that three serine- and two threonine-based proteins, detected in greater concentrations in M. aeruginosa CS-564/01 than CS-555/1, could covalently interact with carbohydrates (OHenderson et al., 2010a, 2010b-linked glycosylation). These proteins have the ability to form numerous localised networks with carbohydrates and cells in the presence of coagulant molecules, thereby providing a good hypothesis to explain the excellent treatment performance observed for M. aeruginosa CS-564/01 previously. It is proposed that the uronic acids in M. aeruginosa CS-564/01 could interact with proteins via glycosylation, explaining why the coagulant demand for this strain remained low despite the high charged carbohydrate concentration. Overall, it is proposed that process performance could be impacted by: (a) physicochemical characteristics and (b) carbohydrate-protein interactions.

Keywords: Acidic carbohydrates; Glycoproteins; Glycosylation; HPAEC-PAD; LC-MS.

MeSH terms

Carbohydrates
Chlorella vulgaris*
Cyanobacteria*
Microcystis*
Plants

Substances

Carbohydrates