Calcium enhances polyhydroxyalkanoate production and promotes selective growth of the polyhydroxyalkanoate-storing biomass in municipal activated sludge

Estévez-Alonso, Ángel; Arias-Buendía, María; Pei, Ruizhe; Veelen, H. Pieter J. van; Loosdrecht, Mark C.M. van; Kleerebezem, Robbert; Werker, Alan

doi:10.1016/j.watres.2022.119259

Cited by 11 publications

(3 citation statements)

References 49 publications

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“…Fluorescence In Situ Hybridization (FISH) was applied using EUB338-I with the sequence 5′ GCT GCC TCC CGT AGG AGT 3′ labeled with Cy5 fluorophore (biomers.net GmbH, Germany). In a parallel study, the microbial community before and after the PHA accumulation using activated sludge from Bath WWTP was profiled by sequencing of the 16S rRNA gene . Using these sequencing results as the database, the coverage of EUB338-I probe was tested in silico by ARB software .…”

Section: Methodsmentioning

confidence: 99%

“…In a parallel study, the microbial community before and after the PHA accumulation using activated sludge from Bath WWTP was profiled by sequencing of the 16S rRNA gene. 33 Using these sequencing results as the database, the coverage of EUB338-I probe was tested in silico by ARB software. 34 EUB338-I probe alone showed high coverage; therefore, EUB338-II and EUB338-III from EUBmix were not applied in the current study.…”

Section: Methodsmentioning

confidence: 99%

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Visualization of Polyhydroxyalkanoate Accumulated in Waste Activated Sludge

Pei

Vicente-Venegas²,

Tomaszewska-Porada³

et al. 2023

Environ. Sci. Technol.

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Polyhydroxyalkanoates (PHAs) can be produced with municipal waste activated sludge from biological wastewater treatment processes. Methods of selective fluorescent staining with confocal laser scanning microscopy (CLSM) were developed and optimized to evaluate the distribution of PHA storage activity in this mixed culture activated sludge microbial communities. Selective staining methods were applied to a municipal activated sludge during pilot scale PHA accumulation in replicate experiments. Visualization of stained flocs revealed that a significant but limited fraction of the biomass was engaged with PHA accumulation. Accumulated PHA granules were furthermore heterogeneously distributed within and between flocs. These observations suggested that the PHA content for the bacteria storing PHAs was significantly higher than the average PHA content measured for the biomass as a whole. Optimized staining methods provided high acuity for imaging of PHA distribution when compared to other methods reported in the literature. Selective staining methods were sufficient to resolve and distinguish between distinctly different morphotypes in the biomass, and these observations of distinctions have interpreted implications for PHA recovery methods. Visualization tools facilitate meaningful insights for advancements of activated sludge processes where systematic observations, as applied in the present work, can reveal underlying details of structure–function relationships.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Visualization of Polyhydroxyalkanoate Accumulated in Waste Activated Sludge

Pei

Vicente-Venegas²,

Tomaszewska-Porada³

et al. 2023

Environ. Sci. Technol.

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“…Probably, the most well-known microbiome for environmental applications is activated sludge. Studies with these organisms under heterotrophic conditions report up to 60 %dcw PHB 13,14 . Nevertheless, to the authors' knowledge, the use of photosynthetic microbiomes enriched with cyanobacteria for PHB production has only been tested in very few studies, including 5,[15][16][17] .…”

Section: Introductionmentioning

confidence: 99%

Bioplastic production by harnessing cyanobacteria-rich microbiomes for perpetual synthesis

Altamira-Algarra,

Lage,

Meléndez

et al. 2023

Preprint

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Departing from the conventional axenic and heterotrophic cultures, our research ventures into unexplored territory by investigating the potential of photosynthetic microbiomes for polyhydroxybutyrate (PHB) production. A cyanobacteria-rich microbiome was harnessed for PHB production in a 3 L photobioreactor under non-sterile conditions. The robust culture achieved up to 28 %dcwPHB over 108 days of alternating growth and accumulation phases. Nile Blue staining and Transmission Electron Microscope visualization successfully confirmed the presence of PHB granules within cyanobacteria cells. Analysis through proton Nuclear Magnetic Resonance further validated the extracted polymer as PHB. In addition, the overexpression of the enzyme PHA synthase throughout the accumulation phase correlated directly with the increased PHB production. Also, gene expression changes suggested that initially, glycogen served as the primary storage compound. However, with prolonged macronutrient stress, there was a shift of the carbon flux towards favoring PHB synthesis. Overall, it was demonstrated for the first time the feasibility of using a phototrophic microbiome to continuous production of PHB in a non-sterile system, contributing to advancing in the field of biopolymer production and offering valuable insights into the metabolic pathways involved.

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