High Polyhydroxyalkanoate Research Articles

The production of polyhydroxyalkanoate by Bacillus licheniformis using sequential mutagenesis and optimization

The purpose of this study was to enhance the production of polyhydroxyalkanoate (PHA) by sequential mutation of Bacillus licheniformis PHAs-007, using UV and N-methyl-N′-nitro-N-nitrosoguanidine (NTG). In addition, the effect of nutrient additions and environmental conditions were optimized to increase the production of PHA. Bacillus licheniformis PHAs-007 produced high amounts of PHA (64.09 ∼ 68.80% of DCW) under both synthetic and renewable substrates. After mutagenesis treatment, mutant M2-12 was selected from 380 strains, based on its high biomass and PHA concentration. The mutant M2-12 gave the highest value of specific growth rate (0.09/h), biomass (22.24 g/L) and PHA content (19.55 g/L) under optimal conditions, consisting of 3% palm oil mill effluent, with no additional trace elements, at 45oC and pH 7. The mutant strain showed higher resistance to substrate concentrations, as well as pH and temperature, than the wild type. The accumulation of PHA was increased by 3.18-fold compared to the wild type, and the production of PHA by the mutant M2-12 was constantly retained over 12 times of cultivation. The mutation and optimization strategy appear to be suitable for producing high density PHA, reducing the medium cost and consequently lowering the production cost. Interestingly, the mutant strain could synthesize the novel PHA copolymers such as 3-hydroxyvalerate and 3-hydroxyhexanoate, which were not produced by the wild type.

Bulking sludge for PHA production: Energy saving and comparative storage capacity with well-settled sludge

Two acetate-fed sequencing batch reactors (SBR) were operated under an aerobic dynamic feeding (ADF) model (SBR #2) and with anaerobic phase before aerobic phase (SBR #1) to select mixed cultures with a high polyhydroxyalkanoates (PHA) storage response. Although kinetic selection based on storage response should bring about a predominance of floc-formers, a bulking sludge with storage response comparable to well-settled sludge was steadily established. An anaerobic phase was introduced before the aerobic phase in the ADF model to improve the sludge settleability (SBR #1), however, due to the consequent increased feast/famine ratio, the performance of SBR #1, in terms of both the maximum PHB (polyhydroxybutyrate) cell content and ΔPHB, was lower than that of SBR #2. SBR #2 gradually reached a steady state while SBR #1 failed suddenly after 50 days of operation. The maximum specific substrate uptake rate and storage rate for the selected bulking sludge were 0.4 Cmol Ac/(Cmol X·hr) and 0.18 Cmol Ac/(Cmol PHB·hr), respectively, resulting a yield of 0.45 Cmol PHB/(Cmol Ac) in SBR #2 in the culture enrichment phase. A maximum PHB content of 53% of total suspended solids and PHB storage rate of 1.36 Cmol Ac/(Cmol PHB·hr) was achieved at 10.2 hr in batch accumulation tests under nitrogen starvation. The results indicated that it was feasible to utilize filamentous bacteria to accumulate PHA with a rate comparable to well-settled sludge. Furthermore, the lower dissolved oxygen demand of filamentous bacteria would save energy required for aeration in the culture enrichment stage.

Recovery of poly(3‐hydroxybutyrate‐co‐3‐hydroxyhexanoate) from Ralstonia eutropha cultures with non‐halogenated solvents

Reduced downstream costs, together with high purity recovery of polyhydroxyalkanoate (PHA), will accelerate the commercialization of high quality PHA-based products. In this work, a process was designed for effective recovery of the copolymer poly(hydroxybutyrate-co-hydroxyhexanoate) (P(HB-co-HHx)) containing high levels of HHx (>15 mol%) from Ralstonia eutropha biomass using non-halogenated solvents. Several non-halogenated solvents (methyl isobutyl ketone, methyl ethyl ketone, and butyl acetate and ethyl acetate) were found to effectively dissolve the polymer. Isoamyl alcohol was found to be not suitable for extraction of polymer. All PHA extractions were performed from both dry and wet cells at volumes ranging from 2 mL to 3 L using a PHA to solvent ratio of 2% (w/v). Ethyl acetate showed both high recovery levels and high product purities (up to 99%) when using dry cells as starting material. Recovery from wet cells, however, eliminates a biomass drying step during the downstream process, potentially saving time and cost. When wet cells were used, methyl isobutyl ketone (MIBK) was shown to be the most favorable solvent for PHA recovery. Purities of up to 99% and total recovery yields of up to 84% from wet cells were reached. During polymer recovery with either MIBK or butyl acetate, fractionation of the extracted PHA occurred, based on the HHx content of the polymer. PHA with higher HHx content (17-30 mol%) remained completely in solution, while polymer with a lower HHx content (11-16 mol%) formed a gel-like phase. All PHA in solution could be precipitated by addition of threefold volumes of n-hexane or n-heptane to unfiltered PHA solutions. Effective recycling of the solvents in this system is predicted due to the large differences in the boiling points between solvent and precipitant. Our findings show that two non-halogenated solvents are good candidates to replace halogenated solvents like chloroform for recovery of high quality PHA.

Development of Bio-PORec® system for polyhydroxyalkanoates (PHA) production and its storage in mixed cultures of palm oil mill effluent (POME)

High PHA production and storage using palm oil mill effluent (POME) was investigated using a laboratory batch Bio-PORec® system under aerobic-feeding conditions. Results showed that maximum PHA was obtained at a specific rate (qp) of 0.343C-mol/C-molh when air was supplied at 20ml/min. The PHA yield was found to be 0.80C-mol/C-mol acetic acid (HAc) at microaerophilic condition and the mass balance calculation showed that PHA production increased up to 15.68±2.15C-mmol/cycle. The experiments showed that short feeding rate, limited requirements for electron acceptors (e.g. O2, NO3) and nutrients (N and P) showed lower tendency of glycogen accumulation and contributed more to PHA productivity.

Utilization of vinasse for production of poly-3-(hydroxybutyrate-co-hydroxyvalerate) by Haloferax mediterranei

Vinasse, a highly polluting waste of the ethanol industry was utilized for the production of polyhydroxyalkanoate (PHA) by the extremely halophilic archaeon, Haloferax mediterranei in shake-flasks. Following pre-treatment through adsorption on activated carbon, 25%-50% (v/v) pre-treated vinasse was utilized leading to 70% maximum accumulation of PHA. Maximum PHA concentration of 19.7 g/l, product yield coefficient (based on total carbohydrates) of 0.87 and 0.21 g/l h volumetric productivity were achieved. Concomitant lowering of BOD5 of pre-treated vinasse by at least 78% and COD by at least 80% was attained at the end of this process. The PHA was recovered by osmotic lysis of the cells and purification by sodium hypochlorite and organic solvents. Through UV–vis spectroscopy, gas chromatography, differential scanning calorimetry and nuclear magnetic resonance spectroscopy, the PHA was identified as poly-3-(hydroxybutyrate-co-hydroxyvalerate). The 3-hydroxyvalerate content was 12.36 mol % (utilizing 25% pre-treated vinasse) and 14.09 mol % (utilizing 50% pre-treated vinasse). High salt concentration in the medium allowed this process without sterile conditions and thus reduction in costs of sterilization can be envisaged. Activated charcoal pre-treatment of vinasse is economical than competing processes such as ultrafiltration of whey, extrusion and enzymatic treatment of rice and corn starch. Without impacting sugar prices, this process can easily be integrated into a distillery that has fermentation equipment and trained personnel. High PHA content, productivity, zero-cost carbon source, low-cost isolation of a high-purity product and potential integration into ethanol manufacturing unit with concomitant wastewater treatment should merit further development of this process to higher scales.

Enhanced Recovery and Purification of P(3HB-co-3HHx) from Recombinant Cupriavidus necator Using Alkaline Digestion Method

A simple, efficient and economical method for the recovery of P(3HB-co-3HHx) was developed using various chemicals and parameters. The initial content of P(3HB-co-3HHx) in bacterial cells was 50-60 wt%, whereas the monomer composition of 3HHx used in this experiments was 3-5 mol%. It was found that sodium hydroxide (NaOH) was the most effective chemical for the recovery of biodegradable polymer. High polyhydroxyalkanoate purity and recovery yield both in the range of 80-90 wt% were obtained when 10-30 mg/ml of cells were incubated in NaOH at the concentration of 0.1 M for 60-180 min at 30 °C and polished using 20 % (v/v) of ethanol.

Improvement of the production of poly(3‐hydroxybutyrate‐co‐3‐hydroxyvalerate‐co‐4‐hydroxybutyrate) terpolyester by manipulating the culture condition

AbstractBACKGROUND: The aim of this work is to enhance the production of terpolyester poly(3‐hydroxybutyrate‐co‐3‐hydroxyvalerate‐co‐4‐hydroxybutyrate) (P(3HB‐co‐3HV‐co‐4HB)) produced by a locally isolated bacterium, Cupriavidus sp. USMAA2‐4. The monomer composition was varied by supplementing different carbon precursors and by manipulating the culture condition through one‐stage cultivation. The effect of C/N ratio and different concentrations of carbon source and precursors were investigated in order to produce higher content of this terpolyester. Although research on this biodegradable polyester is abundant, studies on terpolyester P(3HB‐co‐3HV‐co‐4HB) are still limited.RESULTS: Supplementation of oleic acid in accumulation medium increased the bacterial growth and polyhydroxyalkanoate (PHA) accumulation. It was also shown that medium consisting of assorted carbon precursors at C/N 20 gave relatively high dry cell weight and P(3HB‐co‐3HV‐co‐4HB) content. Various compositions of terpolyester were obtained when the concentration of oleic acid and 4HB precursors were manipulated. The combination of oleic acid with γ‐butyrolactone and 1‐pentanol was found to be the best combination to produce high PHA content (81 wt%). The composition of monomer in P(3HB‐co‐3HV‐co‐4HB) was produced in the range 8–13 mol% for 3HV and 9–24 mol% for 4HB, respectively.CONCLUSIONS: The production of P(3HB‐co‐3HV‐co‐4HB) in shake‐flasks successfully produced 81 wt% of PHA content. This manipulated culture condition can be used at larger scale to provide modeling for the production of terpolyester in a bioreactor. Copyright © 2012 Society of Chemical Industry

Screening and identification of polyhydroxyalkanoates producing bacteria and biochemical characterization of their possible application

Polyhydroxyalkanoates (PHAs) accumulating bacteria were isolated under various selective conditions such as pH, salt concentrations and types of heavy metal. Fifty strains of bacterial isolates were found to belong to Bacillus, Proteus, Pseudomonas, Aeromonas, Alcaligenes and Chromobacterium, based on phenotypical features and genotypic investigation. Only twenty five bacterial isolates were selected and observed for the production of PHAs. Interestingly, bacteria belonging to Firmucutes Bacillus sp. produced a high amount of PHAs. The maximum PHAs were accumulated by B. licheniformis PHA 007 at 68.80% of dry cell weight (DCW). Pseudomonas sp., Aeromonas sp., Alcaligenes sp. and Chromobacterium sp. were recorded to produce a moderate amount of PHAs, varying from 10.00-44.32% of DCW. The enzymatic activity was preliminarily analyzed by the ratio of the clear zone diameter to colony diameter. Bacillus gave the highest ratio of hydrolysis zone which corresponds to the highest hydrolytic enzyme activities. Bacillus licheniformis PHA 007 had the highest lipase and protease activity at 2.1 and 5.1, respectively. However, the highest amylase activity was observed in Bacillus sp. PHA 023 at 1.4. Determination of metabolic characteristics was also investigated to check for their ability to consume a wide range of substrates. Bacillus, Aeromonas sp. and Alcaligenes sp. had great ability to utilize a variety of substrates. To decrease high PHA cost, different sources of cheap substrates were tested for the production of PHAs. Bacillus cereus PHA 008 gave the maximal yield of PHA production (64.09% of DCW) when cultivated in anaerobically treated POME. In addition, the accumulation of PHA copolymers such as 3-hydroxyvalerate and 3-hydroxyhexanoate was also observed in Bacillus and Pseudomomas sp. strain 012 and 045, respectively. Eight of the nine isolates accumulated a significant amount of PHAs when inexpensive carbon sources were used as substrates. Here it varied from 1.69% of DCW by B. licheniformis PHA 007 to 64.09% of DCW by B. cereus PHA 008.

Influence of Substrate Load on Polyhydroxyalkanoates (Pha) Accumulation by Unenriched Mixed Cultures from Excess Sludge Fermentation Liquid

To reduce the production cost of polyhydroxyalkanoates (PHA) and disposal amount of excess sludge simultaneously, volatile fatty acids (VFAs) from alkaline excess sludge fermentation was used as carbon sources to synthesize PHA by unenriched mixed cultures. Released phosphorus and residual ammonium in the fermentative VFAs was controlled by adding magnesium to form struvite precipitation. Four VFAs liquids obtained was used to test the influence of initial carbon load and carbon to nitrogen ratio (C:N ratio) on the VFAs uptake rate, PHA storage rate and biomass growth rate. Results show that higher initial carbon substrate load results in relatively higher VFAs uptake rate and higher PHA storage rate, while higher initial C:N ratio results in relatively lower biomass growth rate from VFAs. VFAs generated from thermophilic alkaline sludge fermentation were a suitable carbon source for PHA production by mixed cultures.

The effect of carbon source supplementation on the production of poly(3‐hydroxybutyrate‐co‐3‐hydroxyvalerate) by Cupriavidus necator

The objective of the present study was to investigate the ability of Cupriavidus necator to produce poly-(3-hydroxybutyrate-co-3-hydroxyvalerate) on various carbon sources in batch cultivation. These results show that C. necator produces poly-3-hydroxybutyrate from single carbon sources. The highest poly-3-hydroxybutyrate (P3HB) content was achieved at growth on fructose in the exponential growth phase. The maximum yield of the P3HV content was obtained when fructose was mixed with acetate. The highest content P3HB-co-3HV was also achieved by C. necator when we supplied C-excess and N- and P-normal conditions. These results indicate that C. necator accumulates high polyhydroxyalkanoates (PHA) content by depleting these elements in the culture medium. Nitrogen and phosphorus limitation has no significant effect on the PHA production, whereas C-excess leads to an increase in PHA formation of up to 92% PHAs of cell dry weight after growth on 5 g/L acetate and 40 g/L fructose.

Screening for polyhydroxyalkanoate (PHA)-producing bacterial strains and comparison of PHA production from various inexpensive carbon sources

A total of 20 different strains were isolated, purified and screened for polyhydroxyalkanoate (PHA) production. PHA-producing strains were screened by Nile blue staining and confirmed by Sudan Black B staining. Strain 1.1 was selected for further analysis due to its high PHA production ability. PHA production was optimized and time profiling was calculated. PHA production on various different cheap carbon sources, i.e., sugar industry waste (fermented mash, molasses, spent wash) and corn oil, was compared. Cell dry weight and PHA content (%) were calculated and compared. The 12.53 g/L is the CDW of bacterial strain when grown in medium containing corn oil. It was found that corn oil at 12.53 g/L medium can serve as a carbon source for bacterial growth, allowing cells to accumulate PHA up to 35.63 %. The PhaC gene was amplified to confirm the genetic basis for the production of PHAs. Moreover, 16S rRNA gene sequence analysis showed that strain 1.1 belongs to Pseudomonas species.

Optimization of growth media components for polyhydroxyalkanoate (PHA) production from organic acids by Ralstonia eutropha

We employed systematic mixture analysis to determine optimal levels of acetate, propionate, and butyrate for cell growth and polyhydroxyalkanoate (PHA) production by Ralstonia eutropha H16. Butyrate was the preferred acid for robust cell growth and high PHA production. The 3-hydroxyvalerate content in the resulting PHA depended on the proportion of propionate initially present in the growth medium. The proportion of acetate dramatically affected the final pH of the growth medium. A model was constructed using our data that predicts the effects of these acids, individually and in combination, on cell dry weight (CDW), PHA content (%CDW), PHA production, 3HV in the polymer, and final culture pH. Cell growth and PHA production improved approximately 1.5-fold over initial conditions when the proportion of butyrate was increased. Optimization of the phosphate buffer content in medium containing higher amounts of butyrate improved cell growth and PHA production more than 4-fold. The validated organic acid mixture analysis model can be used to optimize R. eutropha culture conditions, in order to meet targets for PHA production and/or polymer HV content. By modifying the growth medium made from treated industrial waste, such as palm oil mill effluent, more PHA can be produced.

Acclimating PHA storage capacity of activated sludge with static magnetic fields

The effect of a static magnetic field on the acclimation of activated sludge with high polyhydroxyalkanoate (PHA) storage capacity was evaluated under aerobic dynamic feeding (ADF) conditions. The acclimation processes were carried out in Sequence Batch Reactors (SBRs) with static magnetic field intensities of 42, 21, 7, or 0 mT. Static magnetic exposure significantly influenced the PHA accumulation in activated sludge. PHA content in biomass reached 66% (on a COD basis) at a magnetic field intensity of 21 mT and an organic load of 5.42 g L −1, which was 16% higher than that of the control. Analysis by PCR-DGGE showed that the underlying mechanism might involve a change in the dominant species of microorganism in each reactor in response to the static magnetic field.

Isolation and characterization of a transposon mutant ofPseudomonas fluorescens BM07 enhancing the production of polyhydroxyalkanoic acid but deficient in cold-induced exobiopolymer production

Pseudomonas fluorescens BM07 is known to produce cold-induced exobiopolymer, which is mainly composed of water-insoluble hydrophobic polypeptides (up to 85%) and saccharides (8%), by decreasing the culture temperature down to as low as 10 degrees C. We screened for transposon insertion mutants of P. fluorescens BM07 that were unable to produce the exobiopolymer. Among the eight mutants that showed the deficiency of exobiopolymer and O-lipopolysaccharide, one mutant BM07-59 that had the highest polyhydroxyalkanoates (PHA) production was selected. The transposon inserted gene in BM07-59 was identified as galU. The disruption of the gene galU coded for the putative product, UDP-glucose pyrophosphorylase (GalU), resulted in 1.5-fold more accumulation of PHA compared with the wild-type strain from 70 mM fructose or galactose at 30 degrees C. Electrophoretic analysis of lipopolysaccharide showed that the mutant lacked the O-antigen lipopolysaccharide bands. The glycosyl composition of the lipopolysaccharide produced by the mutant strain was significantly different from that of the wild-type strain. We suggest that the deletion of galU could be a way to shift carbon flux efficiently from exobiopolymer toward PHA in P. fluorescens BM07.

Enrichment of a Mixed Bacterial Culture with a High Polyhydroxyalkanoate Storage Capacity

Polyhydroxyalkanoates (PHAs) are microbial storage polymers that attract interest as bioplastics. PHAs can be produced with open mixed cultures if a suitable enrichment step based on the ecological role of PHA is used. An acetate-fed sequencing batch reactor operated with 1 day biomass residence time and with feast-famine cycles of 12 h was used to enrich a mixed culture of PHA producers. In subsequent fed-batch experiments under growth limiting conditions, the enriched mixed culture produced PHA up to a cellular content of 89 wt % within 7.6 h (average rate of 1.2 g/g/h). The PHA produced from acetate was the homopolymer polyhydroxybutyrate. The culture was dominated by a Gammaproteobacterium that showed little similarity on 16S rRNA level with known bacteria (<90% sequence similarity). The mixed culture process for PHA production does not require aseptic conditions. Waste streams rather than pure substrates could be used as raw materials.

Metabolic modelling of polyhydroxyalkanoate copolymers production by mixed microbial cultures

BackgroundThis paper presents a metabolic model describing the production of polyhydroxyalkanoate (PHA) copolymers in mixed microbial cultures, using mixtures of acetic and propionic acid as carbon source material. Material and energetic balances were established on the basis of previously elucidated metabolic pathways. Equations were derived for the theoretical yields for cell growth and PHA production on mixtures of acetic and propionic acid as functions of the oxidative phosphorylation efficiency, P/O ratio. The oxidative phosphorylation efficiency was estimated from rate measurements, which in turn allowed the estimation of the theoretical yield coefficients.ResultsThe model was validated with experimental data collected in a sequencing batch reactor (SBR) operated under varying feeding conditions: feeding of acetic and propionic acid separately (control experiments), and the feeding of acetic and propionic acid simultaneously. Two different feast and famine culture enrichment strategies were studied: (i) either with acetate or (ii) with propionate as carbon source material. Metabolic flux analysis (MFA) was performed for the different feeding conditions and culture enrichment strategies. Flux balance analysis (FBA) was used to calculate optimal feeding scenarios for high quality PHA polymers production, where it was found that a suitable polymer would be obtained when acetate is fed in excess and the feeding rate of propionate is limited to ~0.17 C-mol/(C-mol.h). The results were compared with published pure culture metabolic studies.ConclusionAcetate was more conducive toward the enrichment of a microbial culture with higher PHA storage fluxes and yields as compared to propionate. The P/O ratio was not only influenced by the selected microbial culture, but also by the carbon substrate fed to each culture, where higher P/O ratio values were consistently observed for acetate than propionate. MFA studies suggest that when mixtures of acetate and propionate are fed to the cultures, the catabolic activity is primarily guaranteed through acetate uptake, and the characteristic P/O ratio of acetate prevails over that of propionate. This study suggests that the PHA production process by mixed microbial cultures has the potential to be comparable or even more favourable than pure cultures.

Physiological–biochemical properties and the ability to synthesize polyhydroxyalkanoates of the glucose-utilizing strain of the hydrogen bacterium Ralstonia eutropha B8562

Physiological-biochemical, genetic, and cultural properties of the glucose-utilizing mutant strain Ralstonia eutropha B8562 have been compared with those of its parent strain R. eutropha B5786. It has been shown that growth characteristics of the strain cultured on glucose as the sole carbon and energy source are comparable with those of the parent strain. Strain B8562 is characterized by high polyhydroxyalkanoate (PHA) yields on different carbon sources (CO(2), fructose, and glucose). PHA accumulation in the strain batch cultured on glucose under nitrogen deficiency reaches 90 %. The major monomer in the PHA is beta-hydroxybutyric acid (more than 99 mol %); the identified minor components are beta-hydroxyvaleric acid (0.25-0.72 mol %) and beta-hydroxyhexanoic acid (0.08-1.5 mol %). The strain is a promising PHA producer on available sugar-containing media with glucose.

Recovery of Polyhydroxyalkanoate from Activated Sludge in an Enhanced Biological Phosphorus Removal Bench-Scale Reactor

A sequencing batch reactor was used to study the possibility of harvesting polyhydroxyalkanoate (PHA) from enhanced biological phosphorus removal (EBPR) processes without compromising treatment quality. Because, in EBPR, the highest PHA concentrations are observed after exposure of the sludge to anaerobic conditions, PHA accumulation was evaluated with collection of waste activated sludge (WAS) at the end of the anaerobic stage, in addition to the traditional removal after the aerobic stage. The system achieved good phosphorus removal, regardless of the point of WAS collection. When sludge was harvested at the end of the anaerobic stage, the PHA content of the sludge ranged from 7 to 16 mg PHA/100 mg mixed liquor volatile suspended solids. Although this level of PHA production is below levels obtained with pure cultures, the demonstrated ability to harvest PHA, while simultaneously satisfying phosphorus removal in an EBPR process, is a key initial step towards of the use of wastewater treatment plants for PHA production.

Biosynthesis and Characterization of Poly(3-hydroxybutyrate-co-3- hydroxyhexanoate) from Palm Oil Products in a Wautersia eutropha Mutant

Palm kernel oil, palm olein, crude palm oil and palm acid oil were used for the synthesis of poly (3-hydroxybutyrate-co-3-hydroxyhexanoate) [P(3HB-co-3HHx)] by a mutant strain of Wautersia eutropha (formerly Ralstonia eutropha) harboring the Aeromonas caviae polyhydroxyalkanoate (PHA) synthase gene. Palm kernel oil was an excellent carbon source for the production of cell biomass and P(3HB-co-3HHx). About 87% (w/w) of the cell dry weight as P(3HB-co-3HHx) was obtained using 5 g palm kernel oil/l. Gravimetric and microscopic analyses further confirmed the high PHA content in the recombinant cells. The molar fraction of 3HHx remained constant at 5 mol % regardless of the type and concentration of palm oil products used. The small amount of 3HHx units was confirmed by 13C NMR analysis. The number average molecular weight (M(n)) of the PHA copolymer produced from the various palm oil products ranged from 27 0000 to 46 0000 Da. The polydispersity was in the range of 2.6-3.9.

High-Efficiency Production of Bioplastics from Biodegradable Organic Solids

Microbial polyhydroxyalkanoates (PHAs) have been extensively studied as environmentally friendly biodegradable thermoplastics. The major obstacle to wide acceptance of PHAs is their high price, mainly attributed to the costs of raw materials and polymer recovery. A large amount of organic solids are discarded from food production and consumption and may be used as carbonaceous raw materials for production of PHAs. A novel technology was investigated at bench-top scale to produce PHAs from food scraps. The harvested cell mass had a high PHA content (72.6% of dry cell mass), the same as obtained from pure glucose and organic acids. The organic solid was first digested in an acidogenic reactor in which about 60% solid was converted to fermentative products, including short-chain fatty acids. The four major acids were acetic, propionic, butyric, and lactic acids at concentrations of 6, 2, 27, and 33 g/L, respectively. The acids were transported through a membrane barrier via molecular diffusion to an airlift bioreactor, where the acids were utilized by an enriched culture of Ralstonia eutropha for PHA synthesis. Purification of fermentative acids was not performed in this molecular diffusion–based integration of acidogenesis and polymerization. By using a dialysis membrane as the barrier, the dry cell mass concentration and PHA content reached 22.7 g/L and 72.6%, respectively. The PHA was a copolymer of poly (3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) with 2.8 mole % of hydroxyvalerate.