Name: Scalable Step-by-Step Approach of Sustainable Bioplastic Production from Food Waste (Video) | JoVE Journal
Uploaded: 2025-07-18T13:40:00.000+00:00
Duration: 8 min 14 s
Description: Learn a scalable method for converting food waste into biodegradable PHA bioplastics using halophilic microbial fermentation and chemical-free recovery. This sustainable approach addresses plastic pollution and organic waste challenges.

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5 minutes

This study was supported by Department of Energy-Office of Energy Efficiency and Renewable Energy (Prime Award No. DEEE0009268) and United States Department of Agriculture National Institute of Food and Agriculture (Award No. 2023-79000-38973).

The authors have nothing to disclose.

VFA production from aAD 
VFA plays a crucial role as key precursor for H. mediterranei in converting food waste into PHA. Therefore, maintaining a high VFA concentration during aAD is essential. Several strategies can be employed to enhance VFA production in aAD25. One effective approach is SRT optimization, with an ideal duration of 8 days or less, which promotes the washout of methanogens while maximizing hydrolysis and acidogenesis, thereby preventing excessive VF...

Plastic pollution and food waste disposal are growing environmental concerns, highlighting the need for innovative approaches that address both challenges through sustainable waste management and material development1. Conventional petroleum-based plastics are major contributors to environmental pollution and microplastic contamination, persisting in landfills and aquatic ecosystems for centuries2. Polyhydroxyalkanoates (PHAs), a class of microbial bioplastics, offer a fully biodegradable alternative with mechanical properties comparable to conventional plastics while ensuring complete degradation in natural environments3. However, the high production costs of PHAs remain a significant barrier to widespread commercial adoption4. One promising approach to overcoming this limitation is leveraging organic waste as a cost-effective carbon source for microbial PHA biosynthesis, as feedstock accounts for approximately 50% of total production expenses in current industrial processes that rely on pure carbon sources such as corn starch or glucose5. By valorizing waste, this approach not only diverts organic material from landfills but also transforms it into a sustainable alternative to petroleum-based plastics, simultaneously addressing waste accumulation and plastic pollution issues.
H. mediterranei, a halophilic microorganism, demonstrates great potential for utilizing waste-derived feedstocks for PHA production due to its ability to thrive in high-salinity conditions (2-5 M NaCl), which naturally prevents contamination by inhibiting the growth of non-halophilic microbes6. This high-salinity environment eliminates the need for strict sterilization in industrial fermentation, reducing operational costs. Unlike traditional bacterial PHA producers, H. mediterranei efficiently metabolizes a diverse range of waste-derived carbon sources and has been reported to accumulate PHA up to 70% of its dry cell weight (DCW) as a stress response mechanism7,8. Additionally, its high-salinity environment enables simple downstream processing, as osmotic shock induced by water immersion effectively disrupts the cells, allowing for PHA release without the need for enzymatic or chemical treatments9. These advantages position H. mediterranei as an ideal candidate for scalable, waste-based bioplastic production.
Existing studies have explored various waste materials, including cheese whey, olive mill wastewater, and agro-industrial residues, for PHA fermentation by H. mediterranei6,10,11,12,13,14,15,16,17,18. The high variability of waste composition presents a significant challenge, requiring different pretreatment strategies, such as enzymatic hydrolysis, acid hydrolysis, or thermal processing, before microbial assimilation11,17,19,20. Here, arrested anaerobic digestion (aAD) provides a scalable solution by converting diverse food waste into a homogeneous volatile fatty acid (VFA)-rich digestate, serving as a direct precursor for PHA synthesis by H. mediterranei21,22. This process enhances feedstock consistency and mitigates substrate variability while improving microbial fermentation efficiency.
Despite its potential, only a few studies have successfully demonstrated this two-stage process integrating aAD and microbial PHA fermentation for general food waste-to-bioplastic conversion by H. mediterranei23,24,25. This study provides a detailed step-by-step protocol covering food waste pretreatment, VFA production by semi-continuous aAD, and pure culture fermentation of H. mediterranei for PHA production. Furthermore, while previous studies have validated water-based PHA extraction from H. mediterranei, none have quantified PHA recovery efficiency and purity using this method11,18. This study addresses this gap by demonstrating the chemical-free PHA recovery method utilizing osmotic lysis with water and quantifying its recovery efficiency in comparison to traditional sodium hypochlorite (NaClO)-based PHA recovery methods26. Additionally, this study assesses the purity of water-recovered PHA, providing a comprehensive evaluation of its effectiveness for scalable bioplastic production.
To bridge the gap between laboratory-scale feasibility and industrial implementation, this protocol is designed for scalability in industrial applications, incorporating pilot-scale aAD and PHA fermentation, along with disc centrifugation for cell separation, a widely used technique in bioprocessing and large-scale microbial fermentation27. By optimizing food waste-to-bioplastic conversion, this method supports the industrialization of large-scale waste valorization, advancing circular bioeconomy initiatives. The integration of waste-derived feedstocks, halophilic fermentation, and an environmentally friendly PHA recovery strategy enables efficient waste-to-biopolymer conversion, reducing reliance on fossil-fuel-based plastics while promoting sustainable bioplastic production.

<table><tbody><tr><td>Air</td><td>Airgas</td><td>AI UZ300</td><td>Used for GC operation</td></tr><tr><td>Air pump</td><td>Cloudray</td><td>Hailea ACO-009D</td><td></td></tr><tr><td>Ammonium chloride</td><td>Acros Organics</td><td>A0396560</td><td></td></tr><tr><td>Autoclave</td><td>STERIS</td><td>AMSCO Lab 250</td><td>Used in ATCC medium preparation</td></tr><tr><td>Balance</td><td>METTLER TOLEDO</td><td>ME204TE/00</td><td></td></tr><tr><td>Blender</td><td>VEVOR</td><td>IB750LV+BLD300+WB-1</td><td></td></tr><tr><td>Bucket</td><td>Lowe&#039;s</td><td>Model 50640</td><td>5-Gallon BPA-Free Plastic General Bucket</td></tr><tr><td>Calcium chloride dihydrate</td><td>Acros Organics</td><td>A0362932</td><td></td></tr><tr><td>Chloroform</td><td>Millipore Sigma</td><td>C2432</td><td></td></tr><tr><td>Desiccator</td><td></td><td></td><td>Any type that can be used in TS, VS, VSS measurement</td></tr><tr><td>Digestion tubes, screw cap with PTFE liner</td><td>Millipore Sigma</td><td>CLS982613</td><td></td></tr><tr><td>Digital Reactor Block</td><td>Hach Company</td><td>DRB200</td><td>Used in Hach VFA quantification methods</td></tr><tr><td>Digital stirrer</td><td>SCILOGEX</td><td>SCI550-Pro</td><td></td></tr><tr><td>Disc centrifuge</td><td>Alfa Laval Inc.</td><td>Clara 20</td><td></td></tr><tr><td>Erlenmeyer flasks</td><td>Amazon</td><td>B07V3JPQMM</td><td></td></tr><tr><td>Ethanol</td><td>Amazon</td><td>B09DGVL4ML</td><td></td></tr><tr><td>Freeze dryer</td><td>Labconco</td><td>10030133, 1027319, and 0109621</td><td>FreeZone 2.5 Liter Freeze Dry Systems</td></tr><tr><td>Fume hood</td><td>Kewaunee Scientific Corporation</td><td>Supreme Air LV</td><td></td></tr><tr><td>Furnace</td><td>Thermo Scientific</td><td>Model F30428C</td><td></td></tr><tr><td>Gas chromatography (GC)</td><td>Agilent</td><td>Model 8890&amp;nbsp;</td><td></td></tr><tr><td>Haloferax mediterranei</td><td>ATCC</td><td>33500</td><td></td></tr><tr><td>Hydrogen</td><td>Airgas</td><td>HY UHP300</td><td>Used for GC operation</td></tr><tr><td>Knife</td><td></td><td></td><td>Any type used for cutting food waste</td></tr><tr><td>Lubricant oil</td><td>Fisher Scientific Company</td><td>NC9844626</td><td></td></tr><tr><td>Magnesium chloride hexahydrate</td><td>Fisher Scientific Company</td><td>BP214-500</td><td></td></tr><tr><td>Magnesium sulfate heptahydrate</td><td>Millipore Sigma</td><td>M1880</td><td></td></tr><tr><td>Methonal</td><td>Millipore Sigma</td><td>34860</td><td></td></tr><tr><td>Methyl benzoate</td><td>Millipore Sigma</td><td>93583</td><td>Used as internal standard in PHA quantification</td></tr><tr><td>Microplate reader&amp;nbsp;</td><td>BioTek Synergy</td><td>Synergy H1</td><td></td></tr><tr><td>Nitrogen</td><td>Airgas</td><td>NI UHP300</td><td>Used for GC operation</td></tr><tr><td>Oven</td><td>Thermo Scientific</td><td>OMH750</td><td></td></tr><tr><td>Peristaltic pump</td><td>Simply Pumps Inc.</td><td>PM6000 AC</td><td>High-flow, self-priming pump with excellent dry-running capability, capable of handling high-particulate slurries and designed for 1/8-inch wall tubing.</td></tr><tr><td>pH meter</td><td>Fisher Scientific Company</td><td>AE150</td><td></td></tr><tr><td>PHA standard</td><td>Millipore Sigma</td><td>403105</td><td></td></tr><tr><td>Pilot aAD digester</td><td>Quasar energy group, Independence, Ohio, USA</td><td>Stainless-steel fermenter with a total volume of 100 L and a working volume of 80 L&amp;nbsp;</td><td></td></tr><tr><td>Pilot PHA fermenter</td><td>manukatree</td><td>SF-50l ISO 9001 CE</td><td>Double Layer Jacketed Glass Reactor</td></tr><tr><td>Potassium Chloride</td><td>G-BIOSCIENCES</td><td>162006</td><td></td></tr><tr><td>Pump tubing</td><td></td><td></td><td>Any brand of silicone tubing with a 1/8&#039;&#039; ID and a 3/8&#039;&#039; OD</td></tr><tr><td>Sodium Bicarbonate</td><td>Fisher Chemical</td><td>241787</td><td></td></tr><tr><td>Sodium Bromide</td><td>Carolina</td><td>21139-45</td><td></td></tr><tr><td>Sodium chloride</td><td>Amazon&amp;nbsp;</td><td>B0787CFHF3</td><td></td></tr><tr><td>Sodium hydroxide, Pellets</td><td>Fisher Bioreagents</td><td>236088</td><td></td></tr><tr><td>Sodium hypochlorite</td><td>Millipore Sigma</td><td>425044</td><td></td></tr><tr><td>Sorvall LYNX 4000 Superspeed Centrifuge</td><td>Thermo Scientific</td><td>75006580</td><td>Used for PHA granule collection in downstream PHA recovery&amp;nbsp;</td></tr><tr><td>Spectrophotometer&amp;nbsp;</td><td>Hach Company</td><td>DR3900</td><td>Used in Hach VFA quantification methods</td></tr><tr><td>Sulfuric acid</td><td>Millipore Sigma</td><td>258105</td><td></td></tr><tr><td>Syringe filter</td><td>Amazon</td><td>B06X6GTKVY</td><td></td></tr><tr><td>TOC analyzer</td><td>Shimadzu</td><td>Model TOC-LCSN</td><td></td></tr><tr><td>Volatile Acids TNTplus Vial Test</td><td>Hach Company</td><td>TNT 872</td><td>Capable to measure VFA in the range of 50 - 2,500 mg/L Acetic Acid.</td></tr><tr><td>Water heater</td><td>Bosch Inc.</td><td>Tronic 3000 T 7-Gallon (ES8)</td><td></td></tr><tr><td>Yeast extract</td><td>Fisher Scientific Company</td><td>AAJ6028736</td><td></td></tr></tbody></table>

scalable step-by-step approach of sustainable bioplastic production from food waste

Scalable Step-by-Step Approach of Sustainable Bioplastic Production from Food Waste

The global microplastic crisis, coupled with the growing challenges of food waste disposal, necessitates innovative solutions to address these environmental issues together. Polyhydroxyalkanoates (PHAs) are unique bioplastics that are fully biodegradable in all environments, including marine ecosystems, offering a sustainable alternative to petroleum-based plastics. At the same time, utilizing food waste as a feedstock for PHA production provides an effective strategy for mitigating the challenges of food waste disposal while producing high-value biodegradable plastics. This study provides a step-by-step protocol for producing PHA from food waste, emphasizing the critical care required to ensure high cellular PHA content and quality. The process begins with arrested anaerobic digestion, which converts food waste into microbially assimilable volatile fatty acids (VFA) by maintaining an anaerobic environment and optimizing parameters, such as solid retention time, to maximize VFA production, a key precursor for PHA synthesis. The VFA-rich digestate is then used to cultivate Haloflex mediterranei, a halophilic microorganism capable of accumulating PHA up to 66% &#177; 5% of its dry cell weight. The high salinity cultivation environment of H. mediterranei prevented culture contamination, ensuring optimal PHA production. Cell growth is monitored by measuring optical density to determine the ideal time for PHA harvesting. Cells are lysed using a chemical-free, water-based method leveraging an osmotic pressure gradient, achieving 93% &#177; 3% PHA recovery, followed by solvent-based PHA purification to obtain a PHA purity of 96% &#177; 2%. Each step is vital to ensure the production of high-quality, biodegradable plastics. This paper provides detailed methods for arrested anaerobic digestion, pure culture fermentation, chemical-free cell lysis, and solvent-based PHA purification, offering a scalable and sustainable approach for converting food waste into biodegradable bioplastics suitable for pilot- and full-scale applications.

Food waste characteristics and VFA production from aAD 
An example of TS, VS, and C:N ratio in prepared food waste slurry is shown in Table 1. Due to the inherent variability in food waste composition across different sampling sources and time, fluctuations in TS, VS, and C:N ratio values are expected. While the variability is acceptable, regular TS, VS, and C:N ratio measurements are essential for process monitoring, microbial balance, and overall aAD system optimization<sup class="x...

This protocol presents a scalable method for converting food waste into biodegradable polyhydroxyalkanoate (PHA) bioplastics. It utilizes arrested anaerobic digestion for food waste pretreatment, halophilic microbial fermentation for PHA biosynthesis, and a chemical-free downstream process for PHA recovery.

The global microplastic crisis, coupled with the growing challenges of food waste disposal, necessitates innovative solutions to address these ...

We develop a scalable process to convert food waste into biodegradable plastic using halophilic microbes, which addresses both plastic pollution and organic waste challenges.Key challenges include managing feedstock variability, scaling up without contamination, and achieving high-purity bioplastic while avoiding expensive or hazardous chemical processes.We achieved 93%recovery of biodegradable plastic from food waste using only water, offering a chemical-free solution for downstream processing.Our work demonstrates a viable path from lab to pilot scale for converting food waste into high-quality bioplastic, paving the way for industrial implementation and the broader circular economy practices.To begin, add 10 kilograms of food waste into a 5-gallon bucket.Pour 2.5 liters of water into the bucket containing the food waste.Connect the blender to a power source.Submerge the blade into the food waste and water mixture.Then press the start button to begin blending.Continue blending for several minutes until the mixture is fully homogenized.Prior to feeding, temporarily increase the digester stirring speed from 150 revolutions per minute to 200 revolutions per minute and maintain this speed for 20 minutes.Then, open the discharge valve at the bottom of the pilot anaerobic acidogenic digester and release approximately 26.7 liters of digestate, which is approximately 1/3 of the digester content.If not used immediately, store the collected digestate at 4

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Learn a scalable method for converting food waste into biodegradable PHA bioplastics using halophilic microbial fermentation and chemical-free recovery. This sustainable approach addresses plastic pollution and organic waste challenges.

Scalable Step-by-Step Approach of Sustainable Bioplastic Production from Food Waste

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