Biofilm mitigation in hybrid chemical-biological upcycling of waste polymers.

Introduction: Accumulation of plastic waste in the environment is a serious global issue. To deal with this, there is a need for improved and more efficient methods for plastic waste recycling. One approach is to depolymerize plastic using pyrolysis or chemical deconstruction followed by microbial-upcycling of the monomers into more valuable products. Microbial consortia may be able to increase stability in response to process perturbations and adapt to diverse carbon sources, but may be more likely to form biofilms that foul process equipment, increasing the challenge of harvesting the cell biomass. Methods: To better understand the relationship between bioprocess conditions, biofilm formation, and ecology within the bioreactor, in this study a previously-enriched microbial consortium (LS1_Calumet) was grown on (1) ammonium hydroxide-depolymerized polyethylene terephthalate (PET) monomers and (2) the pyrolysis products of polyethylene (PE) and polypropylene (PP). Bioreactor temperature, pH, agitation speed, and aeration were varied to determine the conditions that led to the highest production of planktonic biomass and minimal formation of biofilm. The community makeup and diversity in the planktonic and biofilm states were evaluated using 16S rRNA gene amplicon sequencing. Results: Results showed that there was very little microbial growth on the liquid product from pyrolysis under all fermentation conditions. When grown on the chemically-deconstructed PET the highest cell density (0.69 g/L) with minimal biofilm formation was produced at 30°C, pH 7, 100 rpm agitation, and 10 sL/hr airflow. Results from 16S rRNAsequencing showed that the planktonic phase had higher observed diversity than the biofilm, and that Rhodococcus, Paracoccus, and Chelatococcus were the most abundant genera for all process conditions. Biofilm formation by Rhodococcus sp. And Paracoccus sp. Isolates was typically lower than the full microbial community and varied based on the carbon source. Discussion: Ultimately, the results indicate that biofilm formation within the bioreactor can be significantly reduced by optimizing process conditions and using pure cultures or a less diverse community, while maintaining high biomass productivity. The results of this study provide insight into methods for upcycling plastic waste and how process conditions can be used to control the formation of biofilm in bioreactors.

Coexistence of specialist and generalist species within mixed plastic derivative-utilizing microbial communities.

BACKGROUND: Plastic-degrading microbial isolates offer great potential to degrade, transform, and upcycle plastic waste. Tandem chemical and biological processing of plastic wastes has been shown to substantially increase the rates of plastic degradation; however, the focus of this work has been almost entirely on microbial isolates (either bioengineered or naturally occurring). We propose that a microbial community has even greater potential for plastic upcycling. A microbial community has greater metabolic diversity to process mixed plastic waste streams and has built-in functional redundancy for optimal resilience. RESULTS: Here, we used two plastic-derivative degrading communities as a model system to investigate the roles of specialist and generalist species within the microbial communities. These communities were grown on five plastic-derived substrates: pyrolysis treated high-density polyethylene, chemically deconstructed polyethylene terephthalate, disodium terephthalate, terephthalamide, and ethylene glycol. Short-read metagenomic and metatranscriptomic sequencing were performed to evaluate activity of microorganisms in each treatment. Long-read metagenomic sequencing was performed to obtain high-quality metagenome assembled genomes and evaluate division of labor. CONCLUSIONS: Data presented here show that the communities are primarily dominated by Rhodococcus generalists and lower abundance specialists for each of the plastic-derived substrates investigated here, supporting previous research that generalist species dominate batch culture. Additionally, division of labor may be present between Hydrogenophaga terephthalate degrading specialists and lower abundance protocatechuate degrading specialists. Video Abstract.

Deconstructed Plastic Substrate Preferences of Microbial Populations from the Natural Environment.

Over half of the world's plastic waste is landfilled, where it is estimated to take hundreds of years to degrade. Given the continued use and disposal of plastic products, it is vital that we develop fast and effective ways to utilize plastic waste. Here, we explore the potential of tandem chemical and biological processing to process various plastics quickly and effectively. Four samples of compost or sediment were used to set up enrichment cultures grown on mixtures of compounds, including disodium terephthalate and terephthalic acid (monomers of polyethylene terephthalate), compounds derived from the chemical deconstruction of polycarbonate, and pyrolysis oil derived from high-density polyethylene plastics. Established enrichment communities were also grown on individual substrates to investigate the substrate preferences of different taxa. Biomass harvested from the cultures was characterized using 16S rRNA gene amplicon sequencing and shotgun metagenomic sequencing. These data reveal low-diversity microbial communities structured by differences in culture inoculum, culture substrate source plastic type, and time. Microbial populations from the classes Alphaproteobacteria, Gammaproteobacteria, Actinobacteria, and Acidobacteriae were significantly enriched when grown on substrates derived from high-density polyethylene and polycarbonate. The metagenomic data contain abundant aromatic and aliphatic hydrocarbon degradation genes relevant to the biodegradation of deconstructed plastic substrates used here. We show that microbial populations from diverse environments are capable of growth on substrates derived from the chemical deconstruction or pyrolysis of multiple plastic types and that paired chemical and biological processing of plastics should be further developed for industrial applications to manage plastic waste. IMPORTANCE The durability and impermeable nature of plastics have made them a popular material for numerous applications, but these same qualities make plastics difficult to dispose of, resulting in massive amounts of accumulated plastic waste in landfills and the natural environment. Since plastic use and disposal are projected to increase in the future, novel methods to effectively break down and dispose of current and future plastic waste are desperately needed. We show that the products of chemical deconstruction or pyrolysis of plastic can successfully sustain the growth of low-diversity microbial communities. These communities were enriched from multiple environmental sources and are capable of degrading complex xenobiotic carbon compounds. This study demonstrates that tandem chemical and biological processing can be used to degrade multiple types of plastics over a relatively short period of time and may be a future avenue for the mitigation of rapidly accumulating plastic waste.

Metagenomic Sequencing of Two Cultures Grown on Chemically Deconstructed Plastic Products.

We report the metagenome sequences of two microbial cultures that were grown with chemically deconstructed plastic products as their sole carbon source. These metagenomes will provide insights into the metabolic capabilities of cultures grown on deconstructed plastics and can serve as a starting point for the identification of novel plastic degradation mechanisms.

Study of the Viscosity and Thermal Characteristics of Polyolefins/Solvent Mixtures: Applications for Plastic Pyrolysis.

Plastic pollution is one of the biggest environmental problems that the world is currently facing. Pyrolysis is a frontier technique aimed at converting plastic waste back into virgin-quality resin. However, the transfer of the waste plastic feed into the pyrolysis reactor must be optimized before the process can be upscaled to a continuous process. In this study, a new solvent that reduces the viscosity of molten plastic was introduced and characterized. The results revealed that the polymers are soluble in the ratio of up to 75 wt % plastic and 25 wt % solvent at 240 °C. The viscosity of pure low-density polyethylene (LDPE), high-density polyethylene (HDPE), and polypropylene (PP) in the solvent was measured in different weight percentages of polymer in solvent (30-80 wt %) and at 160, 180, 200, 220, 240, and 260 °C. The viscosity decreased with the decreasing polymer-weight percentage and with increasing temperature. The viscosity of LDPE/solvent and PPs(isotactic)/solvent is much lower than for HDPE/solvent and PPp(polypropylene impact copolymer)/solvent. Differential scanning calorimetry (DSC) and thermal gravimetric analysis (TGA) were applied to characterize the thermal behavior of LDPE, HDPE, and PP in the solvent in three different weight percentages (25, 50, and 75 wt %). The DSC results indicate that in the mixture of PPs/solvent and LDPE/solvent the melting point of PP and LDPE decreases as the amount of solvent increases. Overall, these results indicate that the selected solvent is an effective agent to prepare waste plastics for pyrolysis.

On the Method of Pulse-Heated Analysis of Solid Reactions (PHASR) for Polyolefin Pyrolysis.

The continued need for plastics necessitates an effective solution for processing and recycling polymer wastes. While pyrolysis is a promising technology for polyolefin recycling, an experimental apparatus must be designed to measure the intrinsic kinetics and elucidate the chemistry of the plastics pyrolysis process. To resolve this issue, a modified Pulse-Heated Analysis of Solid Reactions (PHASR) system was designed, constructed, and evaluated for the purposes of polyolefin pyrolysis. Experimental results demonstrated that the new PHASR system is capable of measuring the millisecond-resolved evolution of plastic [e. g., low-density polyethylene (LDPE)] pyrolysis products at a constant temperature. The PHASR system was shown to be capable of producing a repeatable, fast heating time (20 ms) and cooling time (130-150 ms), and of maintaining a stable temperature during reaction. A second, Visual PHASR system was developed to enable high-speed photography and visualization of the real-time pyrolysis of LDPE.

Biomass microspheres - A new method for characterization of biomass pyrolysis and shrinkage.

A method for manufacturing biomass microspheres is introduced. Biomass microspheres between 100 and 400 µm in diameter were produced from crystalline cellulose, switchgrass, and tall fescue using spray drying. The biomass microspheres were characterized using X-ray computed tomography (XCT), thermal gravimetric analysis (TGA), and real-time optical imaging: density, pore structure, and size effects were studied. Dynamic shrinkage was video-captured using a CMOS camera and parallel (paired, side-by-side) weight loss experiments were performed using TGA. The combined data was used to correlate the extent of pyrolysis to gross morphological change (shrinkage). The results show that manufactured biomass microspheres are uniform and that, in the absence of heat and mass transport limitations, pyrolysis shrinkage is an intrinsic property dependent only upon the final processing temperatures.