Enhancing the production of PHA in Scenedesmus sp. by the addition of green synthesized nitrogen, phosphorus, and nitrogen-phosphorus-doped carbon dots.

Plastic consumption has increased globally, and environmental issues associated with it have only gotten more severe; as a result, the search for environmentally friendly alternatives has intensified. Polyhydroxyalkanoates (PHA), as biopolymers produced by microalgae, might be an excellent option; however, large-scale production is a relevant barrier that hinders their application. Recently, innovative materials such as carbon dots (CDs) have been explored to enhance PHA production sustainably. This study added green synthesized multi-doped CDs to Scenedesmus sp. microalgae cultures to improve PHA production. Prickly pear was selected as the carbon precursor for the hydrothermally synthesized CDs doped with nitrogen, phosphorous, and nitrogen-phosphorous elements. CDs were characterized by different techniques, such as FTIR, SEM, ζ potential, UV-Vis, and XRD. They exhibited a semi-crystalline structure with high concentrations of carboxylic groups on their surface and other elements, such as copper and phosphorus. A medium without nitrogen and phosphorous was used as a control to compare CDs-enriched mediums. Cultures regarding biomass growth, carbohydrates, lipids, proteins, and PHA content were analyzed. The obtained results demonstrated that CDs-enriched cultures produced higher content of biomass and PHA; CDs-enriched cultures presented an increase of 26.9% in PHA concentration and an increase of 32% in terms of cell growth compared to the standard cultures.

Mechanical, Thermal, and Physicochemical Properties of Filaments of Poly (Lactic Acid), Polyhydroxyalkanoates and Their Blend for Additive Manufacturing.

Polymeric blends are employed in the production of filaments for additive manufacturing to balance mechanical and processability properties. The mechanical and thermal properties of polymeric filaments made of poly (lactic acid) (PLA), polyhydroxyalkanoates (PHA), and its blend (PLA-PHA) are investigated herein and correlated to their measured structural and physicochemical properties. PLA exhibits the highest stiffness and tensile strength, but lower toughness. The mechanical properties of the PLA-PHA blend were similar to those of PLA, but with a significantly higher toughness. Despite the lower mechanical properties of neat PHA, incorporating a small amount (12 wt.%) of PHA into PLA significantly enhances toughness (approximately 50%) compared to pure PLA. The synergistic effect is attributed to the spherulitic morphology of blended PHA in PLA, promoting interactions between the amorphous regions of both polymers. Thermal stability is notably improved in the PLA-PHA blend, as determined by thermogravimetric analysis. The blend also exhibits lower cold crystallization and glass transition temperatures as compared to PLA, which is beneficial for additive manufacturing. Following additive manufacturing, X-ray photoelectron spectroscopic showed that the three filaments present an increase in C-C and C=O bonds associated with the loss of C-O bonds. The thermal process induces a slight increase in crystallinity in PHA due to chain reorganization. The study provides insights into the thermal and structural changes occurring during the melting process of additive manufacturing.

Evaluation of the production and extraction of polyhydroxybutyrate from volatile fatty acids by means of mixed cultures and B. cepacia.

The global consumption of plastics generates accelerated environmental pollution in landfills and marine ecosystems. Biopolymers are the materials with the greatest potential to replace synthetic polymers in the market due to their good biodegradability, however, there are still several disadvantages, mainly related to their production cost. Considering the above, the generation of biodegradable and biocompatible bioplastics stands out as an alternative solution, some of which are made from renewable raw materials, including polyhydroxyalkanoates PHAs. Although much research has been done on bacteria with the capacity for intracellular accumulation of PHAs, among others, it is also possible to produce PHAs using mixed microbial cultures instead of a single microorganism, using natural microbial consortia that have the capacity to store high amounts of PHAs. In this contribution, three methods for the extraction and purification of PHAs produced by fermentation using volatile fatty acids as a carbon source at different concentrations were evaluated, using the pure strain Burkholderia cepacia 2G-57 and the mixed cultures of the activated sludge from the El Salitre WWTP, in order to select the best method from the point of view of environmental sustainability as this will contribute to the scalability of the process. The mixed cultures were identified by sequencing of the 16S gene. A yield of 89% was obtained from the extraction and purification of PHA using acetic acid as a solvent, which according to its properties is "greener" than chloroform. The polymer obtained was identified as polyhydroxybutylated PHB.

Exploring polyhydroxyalkanoates biosynthesis using hydrocarbons as carbon source: a comprehensive review.

Environmental pollution caused by petrochemical hydrocarbons (HC) and plastic waste is a pressing global challenge. However, there is a promising solution in the form of bacteria that possess the ability to degrade HC, making them valuable tools for remediating contaminated environments and effluents. Moreover, some of these bacteria offer far-reaching potential beyond bioremediation, as they can also be utilized to produce polyhydroxyalkanoates (PHAs), a common type of bioplastics. The accumulation of PHAs in bacterial cells is facilitated in environments with high C/N or C/P ratio, which are often found in HC-contaminated environments and effluents. Consequently, some HC-degrading bacteria can be employed to simultaneously produce PHAs and conduct biodegradation processes. Although bacterial bioplastic production has been thoroughly studied, production costs are still too high compared to petroleum-derived plastics. This article aims to provide a comprehensive review of recent scientific advancements concerning the capacity of HC-degrading bacteria to produce PHAs. It will delve into the microbial strains involved and the types of bioplastics generated, as well as the primary pathways for HC biodegradation and PHAs production. In essence, we propose the potential utilization of HC-degrading bacteria as a versatile tool to tackle two major environmental challenges: HC pollution and the accumulation of plastic waste. Through a comprehensive analysis of strengths and weaknesses in this aspect, this review aims to pave the way for future research in this area, with the goal of facilitating and promoting investigation in a field where obtaining PHAs from HC remains a costly and challenging process.

PHB production by Bacillus megaterium strain MNSH1-9K-1 using low-cost media.

Plastics are widely used for diverse applications due to their versatility. However, their negative impact on ecosystems is undeniable due to their long-term degradation. Thus, there is a rising need for developing eco-friendlier alternatives to substitute fossil-based plastics, like biopolymers. PHA are synthesized intracellularly by microorganisms under stressful conditions of growth and have similar characteristics to conventional polymers, like their melting point, transition temperatures, crystallinity, and flexibility. Although it is feasible to use biopolymers for diverse industrial applications, their elevated production cost due to the supplies needed for microbiological procedures and the low productivity yields obtained have been the main limiting factors for their commercial success. The present study assessed the ability of Bacillus megaterium strain MNSH1-9K-1 to produce biopolymers using low-cost media from different kinds of fruit-peel residues. The results show that MNSH1-9K-1 can produce up to 58 g/L of PHB when grown in a medium prepared from orange-peel residues. The data obtained provide information to enhance the scalability of these kinds of biotechnological processes.

Kinetic and stoichiometric parameters in the fed-batch bioreactor production of poly(3-hydroxybutyrate) by Bacillus megaterium using different carbon sources.

This study investigates the effects of different strategies on poly(3-hydroxybutyrate)-P(3HB) production in a fed-batch bioreactor by Bacillus megaterium using candy industry effluent (CIE), sucrose, and rice parboiled water (RPW) as carbon sources. In biosynthesis, kinetic and stoichiometric parameters of substrate conversion into products and/or cells, productivity, instantaneous, and specific conversion rates were evaluated. The maximum concentration of P(3HB) was 4.00 g.L-1 (77% of the total dry mass) in 42 h of cultivation in minimal medium/RPW added with a carbon source based on CIE, demonstrating that the fed-batch provided an increase of approximately 22% in the polymer concentration and 32% in the overall productivity in relation to medium based on commercial sucrose. Fed-batch cultivation also had the advantage of avoiding the extra time required for inoculum preparation and sterilization of the bioreactor during the batch, which thereby increased the overall industrial importance of the process. Effluents from the candy, confectionery, and/or rice parboiling industries can be used as alternative substrates for P(3HB) production at a low cost.

Study of the growth and biochemical composition of 20 species of cyanobacteria cultured in cylindrical photobioreactors.

BACKGROUND: Cyanobacteria are prokaryotic organisms with wide morphological and metabolic diversity. By means of photosynthesis, they convert inorganic compounds into biomolecules, which may have commercial interest. In this work, we evaluated 20 cyanobacterial strains regarding their physiological aspects such as growth, photosynthesis and biochemical composition, some of which are revealed here for the first time. The organisms were cultivated in cylindrical photobioreactors (CPBR) for 144 h and the biomass was obtained. The light inside cultures was constant throughout experimental time and maintained at the saturation irradiance (Ik) of each species. Culture pH was maintained within 7.8 and 8.4 by automatic CO2 bubbling. Growth rate, dry biomass, chlorophyll a, carotenoids, phycocyanin, proteins, carbohydrates, lipids, polyhydroxyalkanoate (PHA) and antioxidant activity were determined. RESULTS: The proportionality of the biochemical composition varied among species, as well as the growth rates. Leptolyngbya sp. and Nostoc sp. (CCIBt3249) showed growth rates in the range of 0.7-0.8 d-1, followed by Rhabdorderma sp. (~ 0.6 d-1), and Phormidium sp. (~ 0.5 d-1). High carotenoid content was obtained in Rhabdoderma sp. (4.0 µg mL-1) and phycocyanin in Leptolyngbya sp. (60 µg mL-1). Higher total proteins were found in the genus Geitlerinema (75% DW), carbohydrates in Microcystis navacekii (30% DW) and lipids in Phormidium sp. (15% DW). Furthermore, Aphanocapsa holsatica showed the highest antioxidant activity (65%) and Sphaerocavum brasiliense, Microcystis aeruginosa, Nostoc sp. (CCIBt3249) and A. holsatica higher levels of PHA (~ 2% DW). CONCLUSIONS: This study reports on the biochemical composition of cyanobacteria that can impact the biotechnology of their production, highlighting potential strains with high productivity of specific biomolecules.

Natural antimicrobial systems protected by complex polyhydroxyalkanoate matrices for food biopackaging applications - A review.

Interest is growing in entrapping natural antimicrobial compounds (NACs) within polyhydroxyalkanoates (PHAs) to produce active food-biopackaging systems. PHAs are versatile polymeric macromolecules that can protect NAC activity by entrapment. This work reviews 75 original papers and 18 patents published in the last 11 years concerning PHAs as matrices for NACs to summarize the physicochemical properties, release, and antimicrobial activities of systems fabricated from PHAs and NACs (PHA/NAC systems). PHA/NAC systems have recently been used as active food biopackaging systems to inactivate foodborne pathogens and prolong food shelf life. PHAs protect NACs by increasing the degradation temperature of some NACs and decreasing their loss of mass when heated. Some NACs also transform the PHA/NAC systems into more thermostable, flexible, and resistant when interacting with PHAs while also improving the barrier properties of the systems. NAC release and activity are also prolonged when NACs are trapped within PHAs. PHA/NAC systems, therefore, represent ecologically friendly materials with promising applications.

Flow cytometry: a tool for understanding the behaviour of polyhydroxyalkanoate accumulators.

The use of mixed microbial cultures (MMCs) is seen as an attractive strategy for polyhydroxyalkanoate (PHA) production. In order to optimize the MMC-PHA production process, tools are required to improve our understanding of the physiological state of the PHA-storing microorganisms within the MMC. In the present study, we explored the use of flow cytometry to analyse the metabolic state and polyhydroxybutyrate (PHB) content of the microorganisms from an MMC-PHA production process. A sequencing batch reactor under a feast and famine regime was used to enrich an MMC with PHB-storing microorganisms. Interestingly, once the PHB-storing microorganisms are selected, the level of PHB accumulation depends largely on the metabolic state of these microorganisms and not exclusively on the consortium composition. These results demonstrate that flow cytometry is a powerful tool to help to understand the PHA storage response of an MMC-PHA production process. KEY POINTS: • Flow cytometry allows to measure PHB content and metabolic activity over time. • Microorganisms showing high PHB content also have high metabolic activity. • PHB producers with low metabolic activity show low PHB content.

PHB Processability and Property Improvement with Linear-Chain Polyester Oligomers Used as Plasticizers.

In 2021, global petroleum-based plastic production reached over 400 million metric tons (Mt), and the accumulation of these non-biodegradable plastics in the environment is a worldwide concern. Polyhydroxybutyrate (PHB) offers many advantages over traditional petroleum-based plastics, being biobased, completely biodegradable, and non-toxic. However, its production and use are still challenging due to its low deformation capacity and narrow processing window. In this work, two linear-chain polyester oligomers were used as plasticizers to improve the processability and properties of PHB. Thermal analyses, XRD, and polarized optical microscopy were performed to evaluate the plasticizing effect on the PHB and the reflection on the mechanical behavior. Both oligomers acted as PHB plasticizers, with a reduction in Tg and Tm as a function of the plasticizer concentration, which can make it easier to handle the material in thermal processing and reduce the probability of thermal degradation. Plasticizer 2 proved to be the most promising between the two with an optimized condition of 20%, in which there was a decrease in elastic modulus of up to 72% and an increase in the maximum elongation of 467%.

Revalorization of Microalgae Biomass for Synergistic Interaction and Sustainable Applications: Bioplastic Generation.

Microalgae and cyanobacteria are photosynthetic microorganisms' sources of renewable biomass that can be used for bioplastic production. These microorganisms have high growth rates, and contrary to other feedstocks, such as land crops, they do not require arable land. In addition, they can be used as feedstock for bioplastic production while not competing with food sources (e.g., corn, wheat, and soy protein). In this study, we review the macromolecules from microalgae and cyanobacteria that can serve for the production of bioplastics, including starch and glycogen, polyhydroxyalkanoates (PHAs), cellulose, polylactic acid (PLA), and triacylglycerols (TAGs). In addition, we focus on the cultivation of microalgae and cyanobacteria for wastewater treatment. This approach would allow reducing nutrient supply for biomass production while treating wastewater. Thus, the combination of wastewater treatment and the production of biomass that can serve as feedstock for bioplastic production is discussed. The comprehensive information provided in this communication would expand the scope of interdisciplinary and translational research.

Aislamiento y caracterización de bacterias productoras de biopolimeros a partir de efluentes industriales / Isolated and characterization of biopolymer producing bacteria from industrial effluents

RESUMEN Se realizó una caracterización físico-química de los efluentes procedentes de industrias del sector educación, metalmecánico, lácteos y confitería de la ciudad de Manizales, Caldas; posteriormente se obtuvieron aislamientos, en medios diferenciales suplementados, de bacterias con potencial para la producción de biopolímeros a los cuales se les aplicó pruebas para la caracterización morfológica, bioquímica y molecular. Los parámetros físico químicos obtenidos de los efluentes industriales demuestran diferencias entre ellos, ya que cada industria genera diferentes residuos aportando una determinada contaminación al efluente, se obtuvieron 73 aislamientos productores de exopolisacáridos (EPS) y 101 productores de polihidroxialcanoatos (PHA), con características morfológicas y bioquímicas variables. El estudio muestra que los efluentes industriales son una gran fuente de bacterias de interés para la producción de diversos polímeros microbianos; principalmente aquellos que producen polímeros tipo biopoliésteres intracelulares como PHA, debido a su variabilidad físico-química y nutricional permitiendo que los microorganismos se adapten a diversas características medioambientales y de composición.

ABSTRACT A physical-chemical characterization of effluents from industries in the education, metal-mechanic, dairy and confectionery sectors of the city of Manizales, Caldas; Later isolates were obtained, in differential media supplemented, from bacteria with potential for the production of biopolymers to which they were applied tests for morphological, biochemical and molecular characterization. The physical chemical parameters obtained from the industrial effluents show a difference between them, since each industry generates different waste contributing a certain contamination to the effluent, 73 isolates producing exopolysaccharides (EPS) and 101 producers of polyhydroxyalkanoates (PHA) were obtained, with morphological characteristics and variable biochemistry. The study shows that industrial effluents are a great source of bacteria of interest for the production of various microbial polymers; mainly those that produce polymers like intracellular biopolyesters such as PHA, due to their physical-chemical and nutritional variability allowing the microorganisms to adapt to diverse environmental and compositional characteristics.

Biopackaging Potential Alternatives: Bioplastic Composites of Polyhydroxyalkanoates and Vegetal Fibers.

Initiatives to reduce plastic waste are currently under development worldwide. As a part of it, the European Union and private and public organizations in several countries are designing and implementing regulations for single-use plastics. For example, by 2030, plastic packaging and food containers must be reusable or recyclable. In another approach, researchers are developing biopolymers using biodegradable thermoplastics, such as polyhydroxyalkanoates (PHAs), to replace fossil derivatives. However, their production capacity, high production costs, and poor mechanical properties hinder the usability of these biopolymers. To overcome these limitations, biomaterials reinforced with natural fibers are acquiring more relevance as the world of bioplastics production is increasing. This review presents an overview of PHA-vegetal fiber composites, the effects of the fiber type, and the production method's impact on the mechanical, thermal, barrier properties, and biodegradability, all relevant for biopackaging. To acknowledge the behaviors and trends of the biomaterials reinforcement field, we searched for granted patents focusing on bio-packaging applications and gained insight into current industry developments and contributions.

Participation of Acyl-Coenzyme A Synthetase FadD4 of Pseudomonas aeruginosa PAO1 in Acyclic Terpene/Fatty Acid Assimilation and Virulence by Lipid A Modification.

The pathogenic bacterium Pseudomonas aeruginosa possesses high metabolic versatility, with its effectiveness to cause infections likely due to its well-regulated genetic content. P. aeruginosa PAO1 has at least six fadD paralogous genes, which have been implicated in fatty acid (FA) degradation and pathogenicity. In this study, we used mutagenesis and a functional approach in P. aeruginosa PAO1 to determine the roles of the fadD4 gene in acyclic terpene (AT) and FA assimilation and on pathogenicity. The results indicate that fadD4 encodes a terpenoyl-CoA synthetase utilized for AT and FA assimilation. Additionally, mutations in fadD paralogs led to the modification of the quorum-sensing las/rhl systems, as well as the content of virulence factors pyocyanin, biofilm, rhamnolipids, lipopolysaccharides (LPS), and polyhydroxyalkanoates. In a Caenorhabditis elegans in vivo pathogenicity model, culture supernatants from the 24-h-grown fadD4 single mutant increased lethality compared to the PAO1 wild-type (WT) strain; however, the double mutants fadD1/fadD2, fadD1/fadD4, and fadD2/fadD4 and single mutant fadD2 increased worm survival. A correlation analysis indicated an interaction between worm death by the PAO1 strain, the fadD4 mutation, and the virulence factor LPS. Fatty acid methyl ester (FAME) analysis of LPS revealed that a proportion of the LPS and FA on lipid A were modified by the fadD4 mutation, suggesting that FadD4 is also involved in the synthesis/degradation and modification of the lipid A component of LPS. LPS isolated from the fadD4 mutant and double mutants fadD1/fadD4 and fadD2/fadD4 showed a differential behavior to induce an increase in body temperature in rats injected with LPS compared to the WT strain or from the fadD1 and fadD2 mutants. In agreement, LPS isolated from the fadD4 mutant and double mutants fadD1/fadD2 and fadD2/fadD4 increased the induction of IL-8 in rat sera, but IL1-ß cytokine levels decreased in the double mutants fadD1/fadD2 and fadD1/fadD4. The results indicate that the fadD genes are implicated in the degree of pathogenicity of P. aeruginosa PAO1 induced by LPS-lipid A, suggesting that FadD4 contributes to the removal of acyl-linked FA from LPS, rendering modification in its immunogenic response associated to Toll-like receptor TLR4. The genetic redundancy of fadD is important for bacterial adaptability and pathogenicity over the host.

Glucose metabolism in Pseudomonas aeruginosa is cyclic when producing Polyhydroxyalkanoates and Rhamnolipids.

Pseudomonas aeruginosa is an important chassis for production of polyhydroxyalkanoates (PHA) and rhamnolipids (RHL). Advances in the understanding of the biosynthesis metabolism of these biocompounds are crucial for increasing yield. 13C-Metabolic Flux Ratio Analysis (13C-MFA) is a technique to estimate in vivo metabolic fluxes ratios. PHA and RHL are essentially non-growth associated products of biotechnological interest and both contain hydroxyalkanoates (HAs), whose labeling patterns could be accessed by GC-MS. In this study, to reveal the relative contributions of the Entner-Doudoroff (ED) pathway and the non-oxidative Pentose Phosphate (PP) pathway to PHA and RHL production, 13C-MFA was performed in Pseudomonas aeruginosa LFM634 when supplied with labeled glucose. This bacterial strain lacks both functional EMP and the oxidative PP branch. Labeling patterns in HAs were measured. Experiments with [U-13C] glucose indicated a low flux though PP pathway. An optimal design of labeling experiment showed that [6-13C] glucose would be the best substrate to enable an estimation of the ED flux with high accuracy. Results of experiments performed with this isotope indicated that about two-thirds of glyceraldehyde 3-phosphate is recycled through a cyclic ED architecture, suggesting that P. aeruginosa utilizes that cycle to regulate the NADPH/Acetyl-CoA ratio for PHA and RHL biosynthesis.

Burkholderia sacchari (synonym Paraburkholderia sacchari): An industrial and versatile bacterial chassis for sustainable biosynthesis of polyhydroxyalkanoates and other bioproducts.

This is the first review presenting and discussing Burkholderia sacchari as a bacterial chassis. B. sacchari is a distinguished polyhydroxyalkanoates producer strain, with low biological risk, reaching high biopolymer yields from sucrose (0.29 g/g), and xylose (0.38 g/g). It has great potential for integration into a biorefinery using residues from biomass, achieving 146 g/L cell dry weight containing 72% polyhydroxyalkanoates. Xylitol (about 70 g/L) and xylonic acid [about 390 g/L, productivity 7.7 g/(L.h)] are produced by the wild-type B. sacchari. Recombinants were constructed to allow the production and monomer composition control of diverse tailor-made polyhydroxyalkanoates, and some applications have been tested. 3-hydroxyvalerate and 3-hydroxyhexanoate yields from substrate reached 80% and 50%, respectively. The genome-scale reconstruction of its metabolic network, associated with the improvement of tools for genetic modification, and metabolic fluxes understanding by future research, will consolidate its potential as a bioproduction chassis.

Beyond PHA: Stimulating intracellular accumulation of added-value compounds in mixed microbial cultures.

This review compiled and analyzed the operational conditions (dissolved oxygen, feast and famine ratio, sequential batch reactor cycle length, organic loading rate (OLR), pH, C/N, and temperature) established during the feast and famine culture strategy for the mixed microbial cultures (MMC) selection to understand how these variables could affect the synthesis of polyhydroxyalkanoates, polyglucose, triacylglycerides, levulinic acid and adipic acid from non-fermented substrates. According to the reported information, the dissolved oxygen has a greater impact on the type and amount of produced compound. In a lesser extent, the OLR and the cycle length were identified to have an impact on the accumulation of polyhydroxyalkanoates, whose accumulation was favored at lower OLR and longer cycle lengths. Thereby, the information of this work will allow the design of future strategies for the simultaneous accumulation of compounds of interest other than the polyhydroxyalkanoates or understand the operational conditions that would optimize the polyhydroxyalkanoates production.

Recent advances in the production of biomedical systems based on polyhydroxyalkanoates and exopolysaccharides.

In recent years, growing attention has been devoted to naturally occurring biological macromolecules and their ensuing application in agriculture, cosmetics, food and pharmaceutical industries. They inherently have antigenicity, low immunogenicity, excellent biocompatibility and cytocompatibility, which are ideal properties for the design of biomedical devices, especially for the controlled delivery of active ingredients in the most diverse contexts. Furthermore, these properties can be modulated by chemical modification via the incorporation of other (macro)molecules in a random or controlled way, aiming at improving their functionality for each specific application. Among the wide variety of natural polymers, microbial polyhydroxyalkanoates (PHAs) and exopolysaccharides (EPS) are often considered for the development of original biomaterials due to their unique physicochemical and biological features. Here, we aim to fullfil a gap on the present associated literature, bringing an up-to-date overview of ongoing research strategies that make use of PHAs (poly (3-hydroxybutyrate), poly (3-hydroxybutyrate-co-3-hydroxyvalerate), poly (3-hydroxyoctanoate), poly(3-hydroxypropionate), poly (3-hydroxyhexanoate-co-3-hydroxyoctanoate), and poly (3-hydroxybutyrate-co-3-hydroxyhexanoate)) and EPS (bacterial cellulose, alginates, curdlan, pullulan, xanthan gum, dextran, hyaluronan, and schizophyllan) as sources of interesting and versatile biomaterials. For the first time, a monograph addressing the properties, pros and cons, status, challenges, and recent progresses regarding the application of these two important classes of biopolymers in biomedicine is presented.

Production and characterization of polyhydroxyalkanoates from industrial waste using soil bacterial isolates.

Nowadays when conventional plastic is being looked as a menace, the possibility of it being replaced with polyhydroxyalkanoates (PHAs) which are biodegradable, environment friendly and biocompatible thermoplastics is not remote. PHAs are a fascinating group of biopolyesters stored within the cytoplasm of numerous bacterial cells as energy and carbon reserves. PHAs signify the best promising biological substitute to certain conventional petrochemical plastics which have wide range of applications in different industries such as biomedical sector, packaging, toners for printing, and adhesives for coating, etc. In the present study, PHAs producing bacterial strains were screened by Sudan black B staining and confirmed by Nile blue A staining. Out of forty bacterial strains showing positive results, six bacterial strains exhibited comparatively higher PHAs production. The highest PHAs producing bacterial strain was identified using 16s rRNA sequencing. Optimization of process parameters was performed by using one factor at a time (OFAT) approach. The isolated bacterium was able to synthesize PHAs when various agro-industrial wastes such as domestic kitchen waste, mixed fruit pulp, sugarcane molasses, and waste flour from bread factory were screened as a carbon substrate in the growth medium. The results showed accumulation of 44.5% PHAs of cell dry weight using domestic kitchen waste as carbon substrate. The characterization of biopolymers was performed using FTIR and XRD analysis. The commercial exploitation of results of this study may serve twin purposes of addressing the challenge of high production cost of PHAs being the major constraint in replacing petro-based plastics as well as address the problem of disposal of recurring domestic kitchen waste and other agro-industrial waste.

Resource recovery in aerobic granular sludge systems: is it feasible or still a long way to go?

Lately, wastewater treatment plants are much often being designed as wastewater-resource factories inserted in circular cities. Among biological treatment technologies, aerobic granular sludge (AGS), considered an evolution of activated sludge (AS), has received great attention regarding its resource recovery potential. This review presents the state-of-the-art concerning the influence of operational parameters on the recovery of alginate-like exopolysaccharides (ALE), tryptophan, phosphorus, and polyhydroxyalkanoates (PHA) from AGS systems. The carbon to nitrogen ratio was identified as a parameter that plays an important role for the optimal production of ALE, tryptophan, and PHA. The sludge retention time effect is more pronounced for the production of ALE and tryptophan. Additionally, salinity levels in the bioreactors can potentially be manipulated to increase ALE and phosphorus yields simultaneously. Some existing knowledge gaps in the scientific literature concerning the recovery of these resources from AGS were also identified. Regarding industrial applications, tryptophan has the longest way to go. On the other hand, ALE production/recovery could be considered the most mature process if we take into account that existing alternatives for phosphorus and PHA production/recovery are optimized for activated sludge rather than granular sludge. Consequently, to maintain the same effectiveness, these processes likely could not be applied to AGS without undergoing some modification. Therefore, investigating to what extent these adaptations are necessary and designing alternatives is essential.