Polyhydroxyalkanoate Synthesis Research Articles

Biosynthesis of Polyhydroxyalkanoates in Cupriavidus necator B-10646 on Saturated Fatty Acids.

It has been established that the wild-type Cupriavidus necator B-10646 strain uses saturated fatty acids (SFAs) for growth and polyhydroxyalkanoate (PHA) synthesis. It uses lauric (12:0), myristic (14:0), palmitic (16:0) and stearic (18:0) acids as carbon sources; moreover, the elongation of the C-chain negatively affects the biomass and PHA yields. When bacteria grow on C12 and C14 fatty acids, the total biomass and PHA yields are comparable up to 7.5 g/L and 75%, respectively, which twice exceed the values that occur on longer C16 and C18 acids. Regardless of the type of SFAs, bacteria synthesize poly(3-hydroxybutyrate), which have a reduced crystallinity (Cx from 40 to 57%) and a molecular weight typical for poly(3-hydroxybutyrate) (P(3HB)) (Mw from 289 to 465 kDa), and obtained polymer samples demonstrate melting and degradation temperatures with a gap of about 100 °C. The ability of bacteria to assimilate SFAs opens up the possibility of attracting the synthesis of PHAs on complex fat-containing substrates, including waste.

Effect of Magnetic Field Enrichment Conditions on the Synthesis of Polyhydroxyalkanoates by Activated Sludge

Effect of Magnetic Field Enrichment Conditions on the Synthesis of Polyhydroxyalkanoates by Activated Sludge

A self-stimulating system based on a polyhydroxyalkanoates coupled induction mechanism and its applications for Halomonas

Inducible gene expression systems are vital tools for synthetic biology. This study focused on a microbial polyhydroxyalkanoate (PHA) related inducible expression system, namely, the regulation mechanism of PHA synthesis associated protein PhaR and PhaP1 in Halomonas bluephagenesis which has employed to enhance PHA synthesis under open unsterile conditions. H. bluephagenesis strains ZS19 and ZS49 with one and two copies of the PHA synthesis system (PphaP1*-phaAB) were constructed based on the PHA self-stimulating synthesis mechanism. When grown in a 7-L bioreactor, H. bluephagenesis ZS19 and ZS49 were grown to 97.4 g/L and 89.6 g/L containing 76.3% and 77.1% PHA in the cell dry weights (CDW), with productivity of 2.06 g/L/h and 2.16 g/L/h, respectively. Compared with its control, their CDW, PHB percent and productivity increased by 20.4% and 10.8%, 13.5% and 14.7 %, and 36.4 % and 43.0 %, respectively. Furthermore, H. bluephagenesis ZS97, engineered with an elongated morphology induced by the PHA-mediated PhaR-phaP1 regulatory system, exhibited a 10 % increase in cell dry weight and a morphology approximately 10 times longer than the wild-type strain, allowing easy separation from the culture broth via centrifugation.

A sustainable synthesis of polyhydroxyalkanoate from stubble waste as a carbon source using Pseudomonas putida MTCC 2475.

Polyhydroxyalkanoates (PHAs) are biodegradable polymers that can be produced from lignocellulosic biomass by microorganisms. Cheap and readily available raw material, such as corn stover waste, has the potential to lessen the cost of PHA synthesis. In this research study, corn stover is pretreated with NaOH under conditions optimized for high cellulose and low lignin with central composite design (CCD) followed by characterization using Fourier-transform infrared spectroscopy (FTIR), thermal gravimetric analysis (TGA), and scanning electron microscopy (SEM). Design expert software performed further optimization of alkali pretreated corn stover for high total reducing sugar (TRS) enhancement using CCD using response surface methodology (RSM). The optimized condition by RSM produced a TRS yield of 707.19mg/g. Fermentation using corn stover hydrolysate by Pseudomonas putida MTCC 2475 gave mcl-PHA detected through gas c hromatography - t andem m ass s pectrometry (GC-MS/MS) and characterization of the PHA film by differential scanning calorimetry (DSC), FTIR, and nuclear magnetic resonance (NMR). Thus, this research paper focuses on using agriculture (stubble) waste as an alternative feedstock for PHA production.

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.

Comparison of batch, fed-batch and continuous operation modes for scalable polyhydroxyalkanoate (PHA) production and carbon sequestration from phenol

Polyhydroxyalkanoates (PHA) are biodegradable intracellular polymers, and a renewable alternative to conventional plastics. To reduce PHA's cost, this study combined an unconventional carbon source, i.e. a toxic aromatics phenol, with an acclimated consortium, to develop a scalable PHA production process. The resultant process performance was systematically compared across three operational modes: batch, fed-batch, and continuous. Phenol toxicity caused inhibition in the batch mode, reducing the PHA synthesis rate but resulting in a high PHA content in cells (PHA% > 50 %). Fed-batch feeding alleviated such inhibition, facilitating a high PHA titer and yield from the substrate. Interestingly, a higher feeding rate in the fed-batch reactor improved its performance, probably by providing a suitable food-to-microbe ratio for the biomass. Using a continuous mode also reduced phenol toxicity, but resulted in more synthesis of non-PHA cellular materials (NPCM). The difference might have been caused by different metabolic states and the presence of intermediates in the reactors. Comparing all 14 conditions under the three modes, feeding phenol more gradually tended to channel substrate away from PHA to NPCM synthesis. The volumetric productivity generally ranged from 3 to 12 mg L−1 min−1, while the biomass productivity was between 2 and 11 mg g initial CDW−1 min−1. A 3-stage, production-harvest and starvation-production process was then designed to simulate real industrialized manufacturing, where the operational modes had a long-term effect on biomass' productivity. Fed-batch mode performed the best, as it fixed 1/3 of phenol's organic carbon in PHA product, suggesting high potential for future application.

Effect of enrichment conditions of secondary feeding on the synthesis of polyhydroxyalkanoates (PHAs) by activated sludge

ABSTRACT Polyhydroxyalkanoates (PHAs) are biodegradable plastics with great performance and development prospects. However, their traditional anaerobic/aerobic enrichment process requires a high concentration of dissolved oxygen (DO), resulting in high energy consumption. In this study, an anaerobic/oxygen-limited with secondary feeding enrichment mode was used to enhance the synthesis of PHAs while reducing energy consumption. The enrichment process of PHAs-synthesizing bacteria lasted up to 100 days, and the experiment was conducted to investigate the change of the PHAs synthesizing ability of the system in this mode by detecting the PHAs content and community distribution of the activated sludge under different stages. Under these conditions, the system enriched two major genera of PHAs-synthesizing bacteria, Thauera (30.21%) and Thiothrix (21.30%). The content of PHAs in the sludge increased from 4.51% to 30.87% and was able to achieve a concomitant increase in poly(3-hydroxyvalerate) (PHV) monomer content. After nitrogen limitation (C/N = 150) treatment, the content of PHAs reached 63.05%. The results showed that the enrichment mode of anaerobic/oxygen-limited with secondary feeding could enrich more PHAs-synthesizing bacteria and significantly increase the synthesis amount of PHAs, which revealed the great potential of this mode in solid waste value-added and reduce the production cost of PHAs and could provide a theoretical basis for the production of PHAs from activated sludge.

Key function of Kouleothrix in stable formation of filamentous aerobic granular sludge at low superficial gas velocity with polymeric substrates

Forming and maintaining stable aerobic granular sludge (AGS) at a low superficial gas velocity (SGV) is challenging, particularly with polymeric substrates. This study cultivated filamentous aerobic granular sludge (FAGS) with filamentous Kouleothrix (Type 1851) at low SGV (0.15 cm/s) utilizing mixed acetate-soluble starch. Within approximately 260 days, notable increases in the relative abundance of Kouleothrix (from 4 % to 10 %) and Ca. Competibacter (from 1 % to 26 %) were observed through 16S rRNA gene analysis. Metagenomic analysis revealed increased expression of functional genes involved in volatile fatty acid (VFA) production (e.g., ackA and pta) and polyhydroxyalkanoate synthesis (e.g., phbB and phbC). Kouleothrix acted as a skeleton for bacterial attachment and was the key fermenting bacteria promoting granulation and maintaining granule stability. This study provides insight into the formation of FAGS with low-energy and non-VFA substrates.

A genetic toolbox to empower Paracoccus pantotrophus DSM 2944 as a metabolically versatile SynBio chassis

BackgroundTo contribute to the discovery of new microbial strains with metabolic and physiological robustness and develop them into successful chasses, Paracoccus pantotrophus DSM 2944, a Gram-negative bacterium from the phylum Alphaproteobacteria and the family Rhodobacteraceae, was chosen. The strain possesses an innate ability to tolerate high salt concentrations. It utilizes diverse substrates, including cheap and renewable feedstocks, such as C1 and C2 compounds. Also, it can consume short-chain alkanes, predominately found in hydrocarbon-rich environments, making it a potential bioremediation agent. The demonstrated metabolic versatility, coupled with the synthesis of the biodegradable polymer polyhydroxyalkanoate, positions this microbial strain as a noteworthy candidate for advancing the principles of a circular bioeconomy.ResultsThe study aims to follow the chassis roadmap, as depicted by Calero and Nikel, and de Lorenzo, to transform wild-type P. pantotrophus DSM 2944 into a proficient SynBio (Synthetic Biology) chassis. The initial findings highlight the antibiotic resistance profile of this prospective SynBio chassis. Subsequently, the best origin of replication (ori) was identified as RK2. In contrast, the non-replicative ori R6K was selected for the development of a suicide plasmid necessary for genome integration or gene deletion. Moreover, when assessing the most effective method for gene transfer, it was observed that conjugation had superior efficiency compared to electroporation, while transformation by heat shock was ineffective. Robust host fitness was demonstrated by stable plasmid maintenance, while standardized gene expression using an array of synthetic promoters could be shown. pEMG-based scarless gene deletion was successfully adapted, allowing gene deletion and integration. The successful integration of a gene cassette for terephthalic acid degradation is showcased. The resulting strain can grow on both monomers of polyethylene terephthalate (PET), with an increased growth rate achieved through adaptive laboratory evolution.ConclusionThe chassis roadmap for the development of P. pantotrophus DSM 2944 into a proficient SynBio chassis was implemented. The presented genetic toolkit allows genome editing and therewith the possibility to exploit Paracoccus for a myriad of applications.

Phenotypic and genotypic characterization of Marinobacterium weihaiense sp. nov. and Marinobacterium marinum sp. nov., isolated from marine sediment, and genomic properties of the genus Marinobacterium.

In this study, two novel bacterial strains were isolated from coastal sediment of Weihai, China. The two strains were Gram-stain-negative and facultatively aerobic, designated 3-1745T and A346T. Based on phenotypic, genetic and phylogenetic properties, strains 3-1745T and A346T represent two novel species of the genus Marinobacterium. The results of genome analysis revealed many central carbohydrate metabolism pathways such as gluconeogenesis, pyruvate oxidation, tricyclic acid cycle, pentose phosphate pathway and PRPP biosynthesis in the genus Marinobacterium. The ability of strains 3-1745T and A346T to utilize volatile fatty acids was experimentally confirmed. Polyhydroxyalkanoate synthases (PhaA, PhaB and PhaC) for the synthesis of polyhydroxyalkanoates were prevalent in the genus Marinobacterium. Multiple BGCs (biosynthetic gene clusters) including betalactone, ectoine, ranthipeptide, redox-cofactor, RiPPs (ribosomally synthesized post-translationally modified peptides) and T3PKS (polyketide synthases) in the genome of the genus Marinobacterium were found. Additional genome analyses suggested that the genus Marinobacterium contained diverse potential mechanisms of salt tolerance and mainly utilized oligosaccharides. This is the first report on broad genomic analyses of the genus Marinobacterium with the description of two novel species and potential ecological and biotechnological implications.

Optimization of activated sludge polyhydroxyalkanoates(PHAs) synthesis system by performing sludge activity recovery experiments and varying the initial sludge concentration

ABSTRACT Polyhydroxyalkanoates(PHAs) are considered a good alternative to petroleum-based plastics because of their good biodegradability and biocompatibility. The synthesis of PHAs using activated sludge can not only solve the problem of the high cost of pure cultures but also improve the utilization value of activated sludge. In this study, sludge activity recovery experiments were firstly conducted and the effects of different initial sludge concentrations on the activated sludge PHAs synthesis system were further investigated. the initial sludge concentrations were 1#SBR (2800 ± 50) mg/L, 2#SBR (4200 ± 50) mg/L, and 3#SBR (5500 ± 50) mg/L. The results showed that the activity, sedimentation performance and PHAs synthesis capacity of activated sludge were enhanced after the sludge activity recovery experiment. At the initial sludge concentration of 4200 mg/L, the activated sludge PHAs synthesis system was operated stably and the synthesis efficiency of PHAs was enhanced. In contrast, at the initial sludge concentration of 2800 and 5500 mg/L, the steady state of the activated sludge PHAs synthesis system was damaged to different degrees at different times, and the synthesis efficiency of PHAs was greatly reduced.

Optimization and Analysis of Polyhydroxyalkanoate (PHA) by Bacillus sp. Strain CL33 and Bacillus flexus Strain S5a from Palm Oil Mill Waste

Polyhydroxyalkanoate (PHA) is a biodegradable polymer that microorganisms can synthesize amidst non-optimal growth conditions with excess carbon sources. Palm oil, rich in fatty acids, serves as a carbon source for PHA synthesis. The bacterial PHA production can be influenced by carbon concentration in the growth medium. Therefore, determining the optimal concentration of palm oil as a carbon source is crucial for PHA production. Additionally, it is possible to determine the type of PHA generated by bacteria, which can then be utilized as information when processing utilizing the PHA. The experiment employed palm oil concentrations of 0.5%, 1%, and 2% and was carried out for periods of 48, 72, 96 hours. It was discovered that Bacillus sp. strain CL33 and Bacillus flexus strain S5a produced the most effective PHA at a concentration of 25 with an incubation period of 96 hours. The PHA generated by these bacteria was quantitatively analyzed through measurements of total bacterial growth, cell dry weight, and the levels of crotonic acid. PHA types were also analyzed using GC-MS, with monomers including 2-hydroxybutyrate(-2HB), 2-hydroxy-3-phenylpropionate (2H3PhP), 3-Hydroxyhexanoate (3HHx), 3-hydroxyoctanoate (3H2O), and 3-hydroxydecanoate (3HD). The Bacillus sp. strain CL33 yielded a PHA level of 92.23%. Meanwhile, Bacillus flexus strain S5a synthesized a polyhydroxyalkanoate comprising mostly 3-hydroxyhexanoate (3HHx) and polydimethylsiloxane (PDMS). The monomers used were decamethyltetrasiloxane, dodecamethylpentasiloxane, hexamethylcyclotrisiloxane, octamethylpentasiloxane, and dodecamethylcyclohexasiloxane. The type of PHA produced accounted for 85.93% of the total.

Ecological succession and community assembly of mixed microbial culture polyhydroxyalkanoate production systems: Drivers of polyhydroxyalkanoate synthesis and Bdellovibrio predation

The production of polyhydroxyalkanoate (PHA) by mixed microbial culture (MMC) can reduce the pollution of plastics. Ecophysiological study of the microbial community assembly and succession is helpful for comprehensive understanding the MMC PHA production process. The operation mode of sequential aerobic dynamic discharge - aerobic dynamic feeding (ADD-ADF) was applied and the operation can be divided into acclimation phase and maturation phase. Deterministic process caused by selective pressure dominated the community assembly throughout the operation. In the acclimation phase, the physical selective pressure recovered the settling capacity of the system, and settling ability of the MMC was closely related to function of PHA synthesis. However, in the maturation phase, stochastic process caused sludge bulking, making the settling ability and PHA synthesis function of the MMC independent on each other. Stochastic process led to the succession of the dominant PHA-producing bacteria, for example, the predation of Paracoccus and Thauera by Bdellovibrio.

Polyhydroxybutyrate (PHB) Biosynthesis by an Engineered Yarrowia lipolytica Strain Using Co-Substrate Strategy

High production cost is one of the major factors that limit the market growth of polyhydroxyalkanoates (PHAs) as a biopolymer. Improving PHA synthesis performance and utilizing low-grade feedstocks are two logical strategies for reducing costs. As an oleaginous yeast, Y. lipolytica has a high carbon flux through acetyl-CoA (the main PHB precursor), which makes it a desired cell factory for PHB biosynthesis. In the current study, two different metabolic pathways (NBC and ABC) were introduced into Y. lipolytica PO1f for synthesizing PHB. Compared to the ABC pathway, the NBC pathway, which includes NphT7 to redirect the lipogenesis pathway and catalyze acetoacetyl-CoA synthesis in a more energy-favored reaction, led to PHB accumulation of up to 11% of cell dry weight (CDW), whereas the ABC pathway resulted in non-detectable accumulations of PHB. Further modifications of the strain with the NBC pathway through peroxisomal compartmentalization and gene dose overexpression reached 41% PHB of CDW and a growth rate of 0.227 h−1. A low growth rate was observed with acetate as the sole source of carbon and energy or glucose as the sole substrate at high concentrations. Using a co-substrate strategy helped overcoming the inhibitory and toxic effects of both substrates. Cultivating the engineered strain in the optimal co-substrate condition predicted by response surface methodology (RSM) led to 83.4 g/L of biomass concentration and 31.7 g/L of PHB. These results offer insight into a more cost-effective production of PHB with engineered Y. lipolytica.

De novo assembly and comparative genome analysis for polyhydroxyalkanoates-producing Bacillus sp. BNPI-92 strain

BackgroundCertain Bacillus species play a vital role in polyhydroxyalkanoate (PHA) production. However, most of these isolates did not properly identify to species level when scientifically had been reported. ResultsFrom NGS analysis, 5719 genes were predicted in the de novo genome assembly. Based on genome annotation using RAST server, 5,527,513 bp sequences were predicted with 5679 bp number of protein-coding sequence. Its genome sequence contains 35.1% and 156 GC content and contigs, respectively. In RAST server analysis, subsystem (43%) and non-subsystem coverage (57%) were generated. Ortho Venn comparative genome analysis indicated that Bacillus sp. BNPI-92 shared 2930 gene cluster (core gene) with B. cereus ATCC 14579 T (AE016877), B. paranthracis Mn5T (MACE01000012), B. thuringiensis ATCC 10792 T (ACNF01000156), and B. antrics Amen T (AE016879) strains. For our strain, the maximum gene cluster (190) was shared with B. cereus ATCC 14579 T (AE016877). For Ortho Venn pair wise analysis, the maximum overlapping gene clusters thresholds have been detected between Bacillus s p.BNPI-92 and Ba. cereus ATCC 14579 T (5414). Average nucleotide identity (ANI) such as OriginalANI and OrthoANI, in silicon digital DND-DNA hybridization (isDDH), Type (Strain) Genome Server (TYGS), and Genome-Genome Distance Calculator (GGDC) were more essentially related Bacillus sp. BNPI-92 with B. cereus ATCC 14579 T strain. Therefore, based on the combination of RAST annotation, OrthoVenn server, ANI and isDDH result Bacillus sp.BNPI-92 strain was strongly confirmed to be a B. cereus type strain. It was designated as B. cereus BNPI-92 strain. In B. cereus BNPI-92 strain whole genome sequence, PHA biosynthesis encoding genes such as phaP, phaQ, phaR (PHA synthesis repressor phaR gene sequence), phaB/phbB, and phaC were predicted on the same operon. These gene clusters were designated as phaPQRBC. However, phaA was located on other operons. ConclusionsThis newly obtained isolate was found to be new a strain based on comparative genomic analysis and it was also observed as a potential candidate for PHA biosynthesis.

Recent Challenges and Trends of Polyhydroxyalkanoate Production by Extremophilic Bacteria Using Renewable Feedstocks.

Polyhydroxyalkanoates (PHAs) are biodegradable polymers with immense potential in addressing the global plastic pollution crisis and advancing sustainable bioplastics production. Among the various microbes known for PHA production, extremophilic bacteria possess unique capabilities to thrive under extreme conditions, making them attractive candidates for PHA synthesis. Furthermore, the utilization of renewable feedstocks for PHA production aligns with the growing demand for sustainable bioplastic alternatives. A diverse range of extremophilic bacteria, especially halophiles and thermophiles, has provided cost-competitive platforms for producing customized PHA polymers. Extremophilic bacteria offer unique advantages over mesophiles due to their contamination resistance, high cell density growth, and unique culture conditions. The current status of Halomonas spp. as a chassis further allows exploration of metabolic engineering approaches to overcome the challenges associated with current industrial biotechnology. This article especially focuses on extremophilic bacteria and explores recent advances in utilizing renewable feedstocks such as lignocellulosic biomass, agro-industrial residues, and waste streams for PHA production. The integration of biorefinery concepts and circular economy principles in PHA manufacturing is also examined. This review is an attempt to provide an understanding of renewable substrates as feedstocks and emerging trends in PHA production by extremophilic bacteria. It underscores the pivotal role of extremophiles and sustainable feedstock sources in advancing the feasibility and eco-friendliness of PHAs as a promising biopolymer alternative.

Heterologous constitutive production of short-chain-length polyhydroxyalkanoates in Pseudomonas putida KT2440: the involvement of IbpA inclusion body protein.

Designing cell factories for the production of novel polyhydroxyalkanoates (PHAs) via smart metabolic engineering is key to obtain à la carte materials with tailored physicochemical properties. To this end, we used the model medium-chain-length-PHA producing bacterium, P. putida KT2440 as a chassis, which is characterized by its metabolic versatility and stress tolerance. Different PHA biosynthetic modules were assembled in expression plasmids using the Golden gate/MoClo modular assembly technique to implement an orthogonal short-chain-lengh-PHA (scl-PHA) switch in a "deaf" PHA mutant. This was specifically constructed to override endogenous multilevel regulation of PHA synthesis in the native strain. We generated a panel of engineered approaches carrying the genes from Rhodospirillum rubrum, Cupriavidus necator and Pseudomonas pseudoalcaligenes, demonstrating that diverse scl-PHAs can be constitutively produced in the chassis strain to varying yields from 23% to 84% PHA/CDW. Co-feeding assays of the most promising engineered strain harboring the PHA machinery from C. necator resulted to a panel of PHBV from 0.6% to 19% C5 monomeric incorporation. Chromosomally integrated PHA machineries with high PhaCCn synthase dosage successfully resulted in 68% PHA/CDW production. Interestingly, an inverse relationship between PhaC synthase dosage and granule size distribution was demonstrated in the heterologous host. In this vein, it is proposed the key involvement of inclusion body protein IbpA to the heterologous production of tailored PHA in P. putida KT2440.

Oxidation of propionate in Pseudomonas sp. LFM046: Relevance to the synthesis of polyhydroxyalkanoates containing odd-chain length monomers and 2-methylisocitrate

Pseudomonas sp. LFM046 produces polyhydroxyalkanoates of medium-chain length. When carbohydrates are used, only even-chain lenght monomers (3HAeven) are generated. Propionate, when used as a co-substrate, enabled the synthesis of odd-chain length monomers, albeit with poor yields (Y3HAodd/prop.) of approx. 10 %. A mini-Tn5 mutant (LFM693) was generated by interrupting the prpB gene, which encodes the 2-methylisocitrate lyase responsible for one step in propionate oxidation via 2-methylcitrate cycle. At low propionate concentrations of 0.1–0.3 g.L−1, LFM693 showed higher Y3HAodd/prop. In contrast, the propionyl incorporation into the polymer was less efficient at high propionate concentrations. The excretion of 2-methylisocitrate partially explained this reduced efficiency. However, mass balances and 13C isotopomer analysis revealed significant propionate oxidation in LFM693 of 21 % and 46 % for propionate concentrations of 0.5 and 1.0 g.L−1, respectively. Experiments using C1-labeled (13C) propionate revealed an alternative propionate oxidation pathway (α-oxidation, β-oxidation, or 3-methylmalate) whose genes are yet to be revealed.

Biotransformation of starch-based wastewater into bioplastics: Optimization of poly(3-hydroxybutyrate) production by Cupriavidus necator DSM 545 using potato wastewater hydrolysate

Biodegradable biopolymers, such as polyhydroxyalkanoates (PHAs), have emerged as an alternative to petrochemical-based plastics. The present work explores the production of PHAs based on the biotransformation of potato processing wastewater and addresses two different strategies for PHA recovery. To this end, culture conditions for PHA synthesis by Cupriavidus necator DSM 545 were optimized on a laboratory scale using a response surface methodology-based experimental design. Optimal conditions rendered a PHB, poly(3-hydroxybutyrate), accumulation of 83.74 ± 2.37 % (5.1 ± 0.2 gL−1), a 1.4-fold increase compared to the initial conditions. Moreover, polymer extraction with non-halogenated agent improved PHB recovery compared to chloroform method (PHB yield up to 78.78 ± 0.57 %), while maintaining PHB purity. (99.83 ± 4.95 %). Overall, the present work demonstrated the potential valorization of starch-based wastewater by biotransformation into PHBs, a high value-added product, and showed that recovery approaches more eco-friendly than the traditional treatments could be applied to PHB recovery to some extent.

Unveiling the mechanism of static magnetic field effects on carbon flux into polyhydroxyalkanoate synthesis by Haloferax mediterranei

The high cost of polyhydroxyalkanoate (PHA) production significantly limits the large-scale industrial production of PHA. In this regard, applying static magnetic field (SMF) to increase PHA productivity could be a novel approach. The study explored the biological magnetic effects of H. mediterranei for PHA production. A magnetic window for PHA synthesis was found under 110 mT SMF, at which the PHA volumetric productivity increased by 27.28% compared with the control group. The carbon flux in the pathway from acetyl-CoA to the PHA synthesis increased while the flux to the TCA cycle decreased. Combined with transcriptome and proteome analysis, the key genes pdhA2, Paaj, and AccA in the PHA synthesis pathway were up-regulated, while gltA in the TCA cycle was down-regulated under 110 mT SMF. This study revealed the mechanism of SMF enhancement on PHA synthesis by H. mediterranei, which provides an effective solution to increasing PHA volumetric productivity.