Maximum Polyhydroxyalkanoates Content Research Articles

Enrichment performance and salt tolerance of polyhydroxyalkanoates (PHAs) producing mixed cultures under different saline environments

The key to the resource recycling of saline wastes in form of polyhydroxyalkanoates (PHA) is to enrich mixed cultures with salt tolerance and PHA synthesis ability. However, the comparison of saline sludge from different sources and the salt tolerance mechanisms of salt-tolerant PHA producers need to be clarified. In this study, three kinds of activated sludge from different salinity environments were selected as the inoculum to enrich salt-tolerant PHA producers under aerobic dynamic feeding (ADF) mode with butyric acid dominated mixed volatile fatty acid as the substrate. The maximum PHA content (PHAm) reached 0.62 ± 0.01, 0.62 ± 0.02, and 0.55 ± 0.03 g PHA/g VSS at salinity of 0.5%, 0.8%, and 1.8%, respectively. Microbial community analysis indicated that Thauera, Paracoccus, and Prosthecobacter were dominant salt-tolerant PHA producers at low salinity, Thauera, NS9_marine, and SM1A02 were dominant salt-tolerant PHA producers at high salinity. High salinity and ADF mode had synergistic effects on selection and enrichment of salt-tolerant PHA producers. Combined correlation network with redundancy analysis indicated that trehalose synthesis genes and betaine related genes had positive correlation with PHAm, while extracellular polymeric substances (EPS) content had negative correlation with PHAm. The compatible solutes accumulation and EPS secretion were the main salt tolerance mechanisms of the PHA producers. Therefore, adding compatible solutes is an effective strategy to improve PHA synthesis in saline environment.

Simultaneous treatment of epichlorohydrin wastewater and polyhydroxyalkanoate recovery by halophilic aerobic granular sludge highly enriched by Halomonas sp.

The treatment of epichlorohydrin (ECH) wastewater exists chances for achieving cleaner production. This study initially employed moderately halophilic aerobic granular sludge (HAGS) to treat ECH wastewater, and the resulting HAGS was utilized to recover polyhydroxyalkanoate (PHA). During the acclimation process of HAGS, the chemical oxygen demand removal efficiency stabilized at 70 %. Moreover, due to the high enrichment of Halomonas sp. (relative abundance of 86 ± 0.50 %), the maximum PHA content of wasted HAGS was 52.67 wt% in the fermentation process. Simultaneously, the utilization of nuclear magnetic resonance spectroscopy (1H and 13C spectra) and fourier transform infrared spectroscopy for the structural analysis of polymers revealed that polyhydroxybutyrate was the predominant substance extracted from HAGS. In this study, the innovative use of highly enriched HAGS for treating ECH wastewater and simultaneously recovering PHA not only enables the efficient biological treatment of ECH wastewater but also realizes resource recovery of ECH wastewater.

Valorization of food waste derived anaerobic digestate into polyhydroxyalkanoate (PHA) using Thauera mechernichensis TL1

Polyhydroxyalkanoate (PHA) is a biopolymer that can be used as a bioplastic, offering a green alternative to petroleum-based plastics. In this study, we investigated PHA production using Thauera mechernichensis TL1. The optimal molar C/N ratio was determined to be 20 from among the ratios of 4, 20, 40, 80, and 200 and in the absence of nitrogen. Food waste anaerobic digestate, mainly comprised of acetate and propionate, was used as the carbon source for PHA production by T. mechernichensis TL1, resulting in a maximum PHA content of 23.98 ± 0.52 wt% (0.52 ± 0.02 g/L PHA) with a PHA productivity of 0.043 g/L-h PHA. In addition, when using acetate and propionate, T. mechernichensis TL1 produced PHA with a maximum PHA content of 57.43 ± 2.84 wt% (2.04 ± 0.11 g/L PHA) and 50.94 ± 1.61 wt% (2.62 ± 0.16 g/L PHA), with a PHA productivity of 0.092 g/L-h PHA and 0.070 g/L-h PHA, respectively. Proton nuclear magnetic resonance spectroscopy (1H NMR) confirmed polyhydroxybutyrate (PHB) production using acetate as a carbon source, and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) production using propionate or food waste anaerobic digestate as the carbon source. The whole-genome analysis of T. mechernichensis TL1 confirmed the existence of a PHA biosynthesis pathway, with the presence of phaA, phaB, phaC (Class I and Class II), and phaJ genes. This study was the first to demonstrate Thauera sp.’s ability to produce PHA from food waste anaerobic digestate, rendering it as a promising candidate for PHA-producing bacteria for the valorization of food waste.

Polyhydroxyalkanoates production by an advanced food-on-demand strategy: The effect of operational conditions

Despite the increasing number of studies related to the polyhydroxyalkanoate (PHA) production from sewage sludge of wastewater treatment plants, there is still a gap in the correlation between the operational conditions, such as the organic loading rate (OLR), and the intracellular polyhydroxyalkanoate (PHA) content, productivity and final recovery of the polymer. Therefore, this work aims to provide experimental data on PHA productivity and purity in view of scaling up the process to an industrial level taking into account process parameters. In view of that, three OLR were applied during the selection of PHA-accumulating bacteria in sewage sludge. Then, the biomass was harvested and subjected to batch accumulation experiments at two organic loads per dosage by employing a tailor-made software to adopt an automated feed-on-demand strategy, which allowed for 30–56 h of accumulation tests in stand-alone mode. Finally, an improved protocol for PHA extraction has been applied. Experimental results show that the maximum PHA content (60% w/w) was achieved using the highest organic load per dosage during the accumulation test with the biomass selected at the highest OLR (1.8 g COD L−1 d−1). Also, the extraction protocol efficiency was proven with four samples with different PHA content, achieving recovery yield as high as 78 ± 3 % with a purity of 89 ± 2 %, thus demonstrating that the adopted strategy might be beneficial for industrial use.

Polyhydroxyalkanoate biosynthesis and optimisation of thermophilic Geobacillus stearothermophilus strain K4E3_SPR_NPP.

Polyhydroxyalkanoates (PHA) can be used to combat the challenges associated with plastic because it is biodegradable and can be produced from renewable resources. Extremophiles are considered to be potential PHA producers. An initial screening for the PHA synthesizing ability of a thermophilic bacteria Geobacillus stearothermophilus strain K4E3_SPR_NPP was carried out using Sudan black B staining. Nile red viable colony staining was used to further verify that the isolates produced PHA. Crotonic acid assays were used to determine the concentrations of PHA. The bacteria showed 31% PHA accumulation per dry cell weight (PHA/DCW) when glucose was used as a carbon source for growth. The molecule was identified to be medium chain length PHA, A copolymer of PHA containing poly(3-hydroxybutyrate)-poly(3-hydroxyvalerate)-poly(3-hydroxyhexanoate) (PHB-PHV-PHHX) using 1H-NMR. Six carbon sources and four nitrogen sources were screened for the synthesis of maximum PHA content, of which lactose and ammonium nitrate showed45% and 53% PHA/DCW respectively. The important factors in the experiment are identified using the Plackett-Burman design, and optimization is performed using the response surface method. Response surface methodology was used to optimize the three important factors, and the maximum biomass and PHA productions were discovered. Optimal concentrations yielded a maximum of 0.48g/l biomass and 0.32g/l PHA, measuring 66.66% PHA accumulation. Dairy industry effluent was employed for the synthesis of PHA, yielding 0.73g/l biomass and 0.33g/l PHA, measuring 45% PHA accumulation. These findings add credibility to the possibility of adopting thermophilic isolates for PHA production using low-cost substrates.

Hazop Analysis of a Bioprocess for Polyhydroxyalkanoate (PHA) Production from Organic Waste: Part B

The production of polyhydroxyalkanoates (PHAs) from industrial waste streams has attracted the attention of researchers and process industries because they could replace traditional plastics. The integrated treatment of civil wastewater along with organic solid wastes in a novel “urban biorefinery” is a very important option to implement a synergic treatment of all relevant bio-waste streams. The development of new biotech processes should consider the occupational health and safety issues from the initial design stages. Among the process hazards analysis techniques, HAZard and OPerability (HAZOP) methodology is widely used for studying both the processes hazards and their operability problems, by exploring the effects of any deviations from design conditions. In the present study, a modified version of HAZOP methodology has been applied to a three-step process developed at pilot scale in the Treviso municipal wastewater treatment plant in order to produce PHA. This paper (part B) shows the results of HAZOP analysis applied to the second process step aimed at culturing the activated sludge under periodic feeding conditions into a sequencing batch reactor (SBR). The analysis applied to the process conditions, corresponding to the maximum PHA content in the biomass, has led to the identification of technical solutions to mitigate the main occupational risks.

Hazop Analysis of a Bioprocess for Polyhydroxyalkanoate (PHA) Production from Organic Waste: Part A

The number of bioprocesses for the circular economy of organic waste has grown in recent years. Implementation of new processes and technologies should consider occupational health and safety issues from the initial design stages. Among the process hazards analysis techniques, HAZard and OPerability (HAZOP) methodology is widely used for studying both the process’s hazards and their operability problems, by exploring the effects of any deviations from design conditions. In the present study, a modified version of HAZOP methodology has been applied to a three-steps process developed at pilot scale in the Treviso municipal wastewater treatment plant in order to produce polyhydroxyalkanoate (PHA) as the final high value product. This paper shows the results of HAZOP analysis applied to the first process step (acidogenic fermentation) aimed at volatile fatty acids production. The analysis has been applied to the process conditions corresponding to the maximum PHA content in the biomass. The HAZOP study results showed that this methodology allowed a comprehensive exploration of conventional chemical engineering process hazards and biological hazards. Final piping and instrumentation diagrams (P&IDs) for acidogenic fermentation have been designed, identifying all prevention measures aimed at managing the hazard and operability issues. The P&ID shows the interconnection of equipment and the instrumentation required for controlling the process.

Strategy for economical and enhanced polyhydroxyalkanoate production from synergistic utilization of palm oil and derived wastewater by activated sludge

The potential of palm oil and derived wastewater pretreated by enzyme as co-substrates to accumulate polyhydroxyalkanoate (PHA) rich in short and medium-chain-length monomers under two feeding strategies was evaluated batchwise through mixed microbial cultures (MMCs) in activated sludge. A terpolymer with the maximum PHA content of 30.5 wt%, volumetric yield of 0.372 g COD/g COD and composition of ca. 84.7 ∼ 97.4/0.5 ∼ 1.6/2.1 ∼ 13.7 (3-hydroxybutyrate/ 3-hydroxyvalerate/ 3-hydroxyoctanoate, %) was achieved as a result of co-substrate incorporation. From the perspective of economic benefits, PHA accumulated via adopting strategy of supplementing carbon source to the same initial concentration per cycle saved 42.7 % of carbon consumption, along with a reduction in culture time (72 h). The above discoveries signify that the combination of palm oil and derived wastewater plus MMCs provides an alternative to the plastics industries for a more sustainable and efficient utilization of biological resources and an economic PHA accumulation approach.

Recirculation factor as a key parameter in continuous-flow biomass selection for polyhydroxyalkanoates production

The effectiveness of polyhydroxyalkanoates (PHA) production with mixed microbial cultures (MMC) largely depends on the selection of PHA-storing microorganisms, conventionally performed in sequencing batch reactors (SBR). These, although easily allow the establishment of the required feast and famine (FF) regime, can represent a factor of cost increase when the process is scaled up. Here, a novel continuous-flow process for MMC selection under FF conditions has been developed by using two sequentially operated reactors. The feast reactor, having a tubular configuration, was continuously fed with a synthetic mixture of acetic and propionic acids (at an organic loading rate of 2.12 gCOD/L d) and the effluent of this reactor was in part sent to the CSTR famine reactor. The recirculation factor (RC), that is the ratio between the recirculation flow rate and the feeding flow rate to the feast reactor, was the main parameter investigated. Four different runs were performed with the RC varying from 1 to 8 and the increase in its value caused a decrease of the biomass residence time in each reactor. The intracellular PHA content in the feast reactor almost linearly increased up to RC 4 (with a value of 34 ± 2 %, wt/wt) and dropped at the RC 8 condition that, however, showed the maximum PHA content (58 ± 5 %, wt/wt) during the accumulation tests. Indeed, the relative abundance of sequences affiliated with putative PHA-storing bacteria increased up to 90.5 % at RC 8 and were dominated by members of the Alphaproteobacteria class mostly represented by the genus Meganema (74 %).

Phenotypic and genomic characterization provide new insights into adaptation to environmental stressors and biotechnological relevance of mangrove Alcaligenes faecalis D334

Alcaligenes faecalis D334 was determined in this study as a salt-tolerant bacterium isolated from mangrove sediment. In response to 6% (w/v) NaCl, strain D334 produced the highest ectoines of 14.14 wt%. To understand adaptive features to mangrove environment, strain D334 was sequenced using Pacific BioScience platform, resulting in a circular chromosome of 4.23 Mb. Of note, D334 genome harbored 81 salt-responsive genes, among which two membrane-associated genes ompc and eric were absent in 3 selected A. faecalis genomes. Apart from that, a complete pathway for ectoine and 5-hydroxyectoine synthesis was predicted. To resist 40 mM H2O2, 46 genetic determinants contributing to oxidative stress response were employed. Moreover, two operons involved in polyhydroxyalkanoate (PHA) production were identified in the D334 genome, resulting in maximum PHA content of 5.03 ± 0.04 wt% and PHA concentration of 0.13 ± 0.001 g/L. A large flagellar biosynthesis operon contributing to swimming motility was found to be conserved in D334 and 8 other A. faecalis genomes. These findings shed light for the first time on the high versatility of A. faecalis D334 genome to adapt to mangrove lifestyle and the possibility to develop D334 as an industrial platform for PHA and 5-hydroxyectoine production.

Valorization of Brewery Waste through Polyhydroxyalkanoates Production Supported by a Metabolic Specialized Microbiome

Simple SummaryRaw brewers’ spent grain, a by-product of beer production, is produced at a large scale and is usually used as animal feed or is landfilled. However, its composition shows that this feedstock has the potential for other applications, such as bioplastics production (e.g., polyhydroxyalkanoates). In this way, the aim of this work was to assess the use of raw brewers’ spent grain for polyhydroxyalkanoates production, adding new value to this feedstock. The results confirm the potential of raw brewers’ spent grain to produce polyhydroxyalkanoates, as the population was enriched in the microorganisms able to accumulate these biopolymers. These results will contribute to society’s knowledge and competence via the development of a treatment process for brewery waste of both environmental (productive waste treatment) and economic interest (production of biopolymers), which will certainly attract its application to the brewery industry worldwide.Raw brewers’ spent grain (BSG), a by-product of beer production and produced at a large scale, presents a composition that has been shown to have potential as feedstock for several biological processes, such as polyhydroxyalkanoates (PHAs) production. Although the high interest in the PHA production from waste, the bioconversion of BSG into PHA using microbial mixed cultures (MMC) has not yet been explored. This study explored the feasibility to produce PHA from BSG through the enrichment of a mixed microbial culture in PHA-storing organisms. The increase in organic loading rate (OLR) was shown to have only a slight influence on the process performance, although a high selectivity in PHA-storing microorganisms accumulation was reached. The culture was enriched on various PHA-storing microorganisms, such as bacteria belonging to the Meganema, Carnobacterium, Leucobacter, and Paracocccus genera. The enrichment process led to specialization of the microbiome, but the high diversity in PHA-storing microorganisms could have contributed to the process stability and efficiency, allowing for achieving a maximum PHA content of 35.2 ± 5.5 wt.% (VSS basis) and a yield of 0.61 ± 0.09 CmmolPHA/CmmolVFA in the accumulation assays. Overall, the production of PHA from fermented BSG is a feasible process confirming the valorization potential of the feedstock through the production of added-value products.

Insights into integration of polyhydroxyalkanoates (PHAs) production into wastewater treatment: Comparison of different electron acceptors on system function and PHA-producer enrichment

Mixed microbial culture polyhydroxyalkanoates (MMC PHA) process combined with mainstream wastewater treatment is a competitive resource recovery integrated process in which economical non-oxygen electron acceptors can be used to enrich PHA-producer and reduce operating costs. Oxygen and three non-oxygen electron acceptors (endogenous nitrate, exogenous nitrate, and nitrite) were used and comprehensively compared in terms of PHA production ability and microbial community ecophysiology in this study. The results showed that the PHA production ability with exogenous nitrate was comparable to that of oxygen, which had a maximum PHA content of 64.0 wt% (versus 62.3% of oxygen), qPHA of 0.7 mg COD/mg Xa/h, as well as 99.1% denitrification efficiency. The non-oxygen acceptors delayed the time for Arcobacter to occupy the dominant niche in the community, and the environmental selection pressure produced was slightly lower than that of oxygen. The symbiotic community between denitrifying bacteria and PHA-producers ensured the high PHA production potential and denitrification efficiency of the non-oxygen system. An economic assessment showed that the integrated PHA production and nitrate removal process with the mainstream wastewater treatment process proposed in this study could reduce aeration energy by 64%, and PHA recovery yield was 0.1 kg CODPHA/kg CODWAS from waste activated sludge (WAS).

Exploring the Limits of Polyhydroxyalkanoate Production by Municipal Activated Sludge.

Municipal activated sludge can be used for polyhydroxyalkanoate (PHA) production, when supplied with volatile fatty acids. In this work, standardized PHA accumulation assays were performed with different activated sludge to determine (1) the maximum biomass PHA content, (2) the degree of enrichment (or volume-to-volume ratio of PHA-accumulating bacteria with respect to the total biomass), and (3) the average PHA content in the PHA-storing biomass fraction. The maximum attained biomass PHA content with different activated sludge ranged from 0.18 to 0.42 gPHA/gVSS, and the degree of enrichment ranged from 0.16 to 0.51 volume/volume. The average PHA content within the PHA-accumulating biomass fraction was relatively constant and independent of activated sludge source, with an average value of 0.58 ± 0.07 gPHA/gVSS. The degree of enrichment for PHA-accumulating bacteria was identified as the key factor to maximize PHA content when municipal activated sludge is directly used for PHA accumulation. Future optimization should focus on obtaining a higher degree of enrichment of PHA-accumulating biomass, either through selection during wastewater treatment or by selective growth during PHA accumulation. A PHA content in the order of 0.6 g PHA/g VSS is a realistic target to be achieved when using municipal activated sludge for PHA production.

Integrated production of biopolymers with industrial wastewater treatment: Effects of OLR on process yields, biopolymers characteristics and mixed microbial community enrichment

The production of polyhydroxyalkanoates (PHA) using industrial wastewaters as feedstocks is a current and challenging topic. This study investigated the production of biopolymers by a mixed microbial culture under different OLRs equal to 1 kgCOD m−3d−1 (Period 1), 2 kgCOD m−3d−1 (Period 2) and 3 kgCOD m−3d−1 (Period 3). The maximum PHA content was achieved in Period 2 (0.38 gPHA gTSS−1), whereas lower values were obtained in Period 1 (0.13 gPHA gTSS−1) and Period 3 (0.26 gPHA gTSS−1). Overall, the maximum PHA productivity resulted equal to 0.08 gPHA L−1 h−1 (P2), 0.05 gPHA L−1 h−1 (P1) and 0.04 gPHA L−1 h−1 (P3), respectively.The molecular weight of the PHA increased from Period 1 (250 kDa) to Period 2 (417 KDa) and Period 3 (463 KDa), although resulting in a slight decrease of crystallinity degree. Microbial community analysis revealed a reduction in bacterial diversity and a progressive shift of the microbial community with the increasing OLR. Alpha-diversity indexes based on Operational Taxonomic Units (OTUs) at 99% identity revealed higher species richness (Taxa (S) 280) and diversity (Shannon (H) 4,06) in Period 1, whereas Period 3 was characterized by reduced richness and diversity and higher dominance (Taxa (S) 133, Shannon (H) 2,40).Based on the results obtained, it was pointed out that the OLR variation determined significant effects on the process performances, as well as on the productivity and quality of the biopolymers. This means that OLR is a key control parameter to maximize the PHA production and control the physical-chemical characteristics of the polymers.

Engineered Mutants of a Marine Photosynthetic Purple Nonsulfur Bacterium with Increased Volumetric Productivity of Polyhydroxyalkanoate Bioplastics.

Polyhydroxyalkanoates (PHAs) are green and sustainable bioplastics that could replace petrochemical synthetic plastics without posing environmental threats to living organisms. In addition, sustainable PHA production could be achieved using marine photosynthetic purple nonsulfur bacteria (PNSBs) that utilize natural seawater, sunlight, carbon dioxide gas, and nitrogen gas for growth. However, PHA production using marine photosynthetic PNSBs has not been economically feasible yet due to its high cost and low productivity. In this work, strain improvement, using genome-wide mutagenesis coupled with high-throughput screening via fluorescence-activated cell sorting, we were able to create Rhodovulum sulfidophilum mutants with enhanced volumetric PHA productivity, with an up to 1.7-fold increase. The best selected mutants (E6 and E6M4) reached the stationary growth phase 1 day faster and accumulated the maximum PHA content 2 days faster than the wild type. Maximizing volumetric PHA productivity before the stationary growth phase is indeed an additional advantage for R. sulfidophilum as a growth-associated PHA producer.

Ecological insight into incompatibility between polymer storage and floc settling in polyhydroxyalkanoate producer selection using complex carbon sources

Polyhydroxyalkanoate (PHA) producer selection is a key step in mixed culture (MC) production. This study focused on incompatibility between PHA storage and floc settling of MCs in the selection process. In a selector using fermented waste activated sludge as substrate under varying organic loading, average maximum PHA content obtained in batch assays increased by ∼ 22 wt% and biomass concentration increased by ∼ 34% with the increasing of organic loading. However, poor floc settling occurred, causing decreased batch PHA production and costly downstream process. A flank community which can corporately use non-VFA organics existed in the selector. When organic loading increased, PHA producers had stronger negative interactions, but not cooperation with the flank community members. Thus, high PHA storage of MCs was bounded to the domination of core PHA producer. But the domination of Thauera bacteria under high organic loading indirectly induced a bloom of filamentous bacteria.

An integrated process for mixed culture production of 3-hydroxyhexanoate-rich polyhydroxyalkanoates from fruit waste

Fermented wastes have been demonstrated to be adequate sources for polyhydroxyalkanoates (PHA) production using mixed microbial cultures (MMC). In general, MMC processes are focused on the production of co-polymers containing 3-hydroxybutyrate (HB) and 3-hydroxyvalerate (HV), which limits the range of polymer properties and aplications. This study aimed at demonstrating the feasibility of producing a ter-polymer enriched in 3-hydroxyhexanoate (HHx) from a waste using a mixed-culture 3-stage process. For this purpose, each of the stages (acidogenic reactor, selection of a PHA-accumulating culture and PHA accumulation) was fine-tuned to maximise the process’s yield and productivity. In the acidogenic reactor, operating conditions were manipulated to ferment fruit waste into a caproate-rich effluent (69%) with a fermentation products’ concentration (FP) of 13.0 gCOD L−1. A culture enriched in a highly efficient PHA-accumulating culture capable of converting caproate into HHx was selected. This culture was used as inoculum and fed with caproate-rich effluent, achieving a maximum PHA content of 71.3%, a PHA productivity of 3.29 gCOD L−1h−1 and a terpolymer with a composition of 33/1/66 (HB/HV/HHx,% wt.). Batch assays were carried out to demonstrate that caproate was the precursor of HHx, and acclimatization of the biomass to caproate was an important factor in the selection step to foster HHx-containing polymer production. This paper reports unprecedented values of yield, productivity and HHx fraction for the production of a mcl-PHA using MC and a fermented waste. These results are a significant contribution towards upscaling the process to convert a waste into HHx-rich PHA.

Polyhydroxyalkanoates and biochar from green macroalgal Ulva sp. biomass subcritical hydrolysates: Process optimization and a priori economic and greenhouse emissions break-even analysis

Although macroalgae biomass is an emerging sustainable feedstock for biorefineries, the optimum process parameters for their hydrolysis and fermentation are still not known. In the present study, the simultaneous production of polyhydroxyalkanoates (PHA) and biochar from green macroalgae Ulva sp. is examined, applying subcritical water hydrolysis and Haloferax mediterranei fermentation. First, the effects of temperature, treatment time, salinity, and solid load on the biomass and PHA productivity were optimized following the Taguchi method. Hydrolysis at 170 °C, 20 min residence time, 38 g L−1 salinity with a seaweed solid load of 5% led to the maximum PHA yield of 0.104 g g−1Ulva and a biochar yield of 0.194 ± 1.23 g g−1Ulva. Second, the effect of different initial culture densities on the biomass and PHA productivity was studied. An initial culture density of 50 g L−1 led to the maximum volumetric PHA productivity of 0.024 ± 0.002 g L−1 h−1 with a maximum PHA content of 49.38 ± 0.3% w/w Sensitivity analysis shows that within 90% confidence, the annual PHA production from Ulva sp. is 148.14 g PHA m−2 year−1 with an annual biochar production of 42.6 g m−2 year−1. Priori economic and greenhouse gas break-even analyses of the process were done to estimate annual revenues and allowable greenhouse gas emissions. The study illustrates that PHA production from seaweed hydrolysate using extreme halophiles coupled to biochar production could become a benign and promising step in a marine biorefinery.

Production and characterization of poly 3-hydroxybutyrate-co-3-hydroxyvalerate in wheat starch wastewater and its potential for nanoparticle synthesis.

Polyhydroxyalkanoates (PHAs) are polymers with biodegradable and biocompatible properties accumulated in a wide variety of bacterial strains. In the present study, active sludge, wheat starch wastewater (WSW), and oil wastewater were used for the isolation and screening of PHA-accumulating bacteria. WSW was then implemented as a cheap and economical culture medium for the production of PHAs by the selected isolate. The extracted PHA was characterized, and the capability of produced biopolymer for preparing nanoparticles was evaluated. Based on the results, 96 different bacterial isolates were obtained, of which the strains isolated from WSW demonstrated the highest PHA-accumulation capability. The maximum PHA content of 3.07g/l (59.50% of dry cell weight) was obtained by strain N6 in 21h. The selected strain was identified by molecular approaches as Bacillus cereus. Afterward, the physicochemical characterization of an accumulated biopolymer was specified as a PHBV copolymer. Finally, spherical homogenous PHBV nanoparticles with a size of 137nm were achieved. The PHBV nanoparticles showed a suitable small size and good zeta potential for medical applications. Hence, it can be concluded that isolated wild strain (B. cereus) has the potential exploitation capability for cost-effective PHBV production using the WSW.

Effects of Various Stress Conditions to Enhance Polyhydroxyalkanoates Accumulation in Wickerhamomyces anomalus VIT-NN01

Background: Polyhydroxyalkanoates (PHA) are polyesters produced by various microorganisms. The major drawback of PHA, which is restricting its large-scale production in industries, is its high cost of raw materials and lower PHA concentration. Objective: The present study aimed to enhance the production of PHA from Wickerhamomyces anomalus VIT-NN01 by optimizing various nutrient and environmental stress conditions and its characterization. Methods: Effect of various stress conditions viz. nitrogen limitation, salinity, chemical mutagens (acridine orange, sodium azide), and physical stresses (UV, Low Electric Current (LEC), Magnetic Field Intensity (MFI), sound waves) were optimized to screen the best strategic growth conditions for the maximum accumulation of PHA in W. anomalus VIT-NN01. Instrumental analysis was done to evaluate the various changes that occurred in the treated cells and extracted PHA. Results: The maximum PHA content was observed on the effect of sound waves (88.74%), followed by LEC (87.8%) and MFI (85.75%). The morphological changes in length, shape, and size of the treated W. anomalus cells were observed by Transmission Electron Microscopy (TEM). Smooth, porous matrix, and pseudo spherical microstructure of the extracted PHA were observed by scanning electron microscopy and TEM analysis. The extracted polymer was identified as poly(3-hydroxybutyrate-co-3- hydroxyvalerate) [P(3HB-co-3HV)] co-polymer comprised of 51.66% 3HB and 48.33% 3HV monomer units based on gas chromatography-mass spectrophotometer and nuclear magnetic resonance spectroscopic analysis. X-ray diffraction analysis revealed the crystalline nature of the extracted P(3HB-co- 3HV). The degradation and melting temperatures were found to be 275.9 and 113.8°C, respectively, through thermogravimetric and differential scanning calorimetry analysis. Conclusion: These results supported the potentiality of W. anomalus, which tolerated the stress conditions and enhanced P(3HB-co-3HV) production from 60 to 88.74% and showed the novelty of present work. This is the first report elucidating the importance of physical stress conditions viz. low electric current, magnetic field intensity, and sound waves for the significant enhancement of PHA production in yeast.