Spatial separation of nitrifiers and denitrifiers promotes selection and enrichment of polyhydroxyalkanoates storing mixed cultures fed by crude glycerol and propionate wastewater.

Polyhydroxyalkanoates (PHA) recovery from industrial wastewater has been highlighted as a promising strategy for a circular bioeconomy. However, the high and varying level of nitrogen in wastewater makes enrichment of mixed microbial culture (MMC) low efficiency. In this study, spatial separation of nitrifiers and denitrifiers was adopted by adding biocarriers in MMC and decreasing the sludge retention time (SRT) to accelerate the enrichment of PHA-storing MMC fed by mixed wastewater containing glycerol and propionate. Nitrifiers and denitrifiers were sustained on biocarriers, obtaining a high total inorganic nitrogen removal and allowing a more efficient selective pressure of a high carbon and nitrogen ratio (C/N) under low SRT conditions. The maximum PHA cell content and relative abundance of PHA-storing bacteria were increased to 60.51 % (SRT 6 d) and 49.62 % (SRT 6 d) with the decrease of SRT, respectively. This study demonstrates an efficient way to highly enrich PHA-storing MMC from crude glycerol, which provide a relevant technical support for high-efficiency enrichment of PHA-storing bacteria in low C/N wastewater.

Mutualistic symbiosis of fungi and nitrogen-fixing bacteria in halophilic aerobic granular sludge treating nitrogen-deficient hypersaline organic wastewater.

Hypersaline organic wastewater is characterized as being nitrogen-deficient, and is easily prone to sludge bulking. In this study, the stability of halophilic aerobic granular sludge (HAGS) for the treatment of hypersaline organic wastewater is explored. Along with the decrease of influent ammonium, the bacterial population substantially reduced, whereas the fungal population continuously increased in HAGS. Saccharomycetales in fungi become the dominant sequence (99.78%) in HAGS bulking. Additionally, Halanaerobium (77.47%) remained prevalent in HAGS despite bacterial washout. Halanaerobium, a nitrogen-fixing genus of bacteria, provided nitrogen for ammonium-assimilating fungi. Saccharomycetales encapsulating HAGS reduced the transfer efficiency of dissolved oxygen, thereby creating favorable growth conditions for Halanaerobium. This paper for the first time highlights the mutualistic symbiosis of fungi and bacteria in HAGS treating the hypersaline organic wastewater. The study lays the foundation for the control and recovery of HAGS bulking.

Simultaneous inorganic nitrogen and phosphate removal by aerobic-heterotrophic fungus Fusarium keratoplasticum FSP1: Performance, pathway and application.

Fungi with multiple contaminant removal function have rarely been studied. Here, a novel fungal strain Fusarium keratoplasticum FSP1, which was isolated from halophilic granular sludge, is reported for first time to perform simultaneous nitrogen and phosphate removal. The strain showed wide adaptability under C/N ratios of 30-35, salinities of 0 %-3 % (m/v), and pH of 7.5-9.5. The maximum removal rates of ammonium, nitrate and nitrite were 4.43, 4.01 and 2.97 mg N/L/h. The nitrogen balance, enzyme activity and substrate conversion experiments demonstrated a single strain FSP1 can assimilate inorganic nitrogen and convert inorganic nitrogen to gaseous nitrogen through heterotrophic nitrification or aerobic denitrification. About 39 %-42 % of the degraded phosphorus was in the extracellular polymeric substances (EPS). Orthophosphate was the main phosphorus species in the cell, whereas phosphate monoester and diester were in the EPS. The novel strain FSP1 is a potential candidate for wastewater treatment.

Feast/famine ratio regulates the succession of heterotrophic nitrification-aerobic denitrification and autotrophic ammonia oxidizing bacteria in halophilic aerobic granular sludge treating saline wastewater.

Heterotrophic nitrification-aerobic denitrification (HN-AD) shows innovation potential of wastewater treatment process in a single tank. However, how to enrich HN-AD bacteria in activated sludge to enhance their contribution remained unknown. This study explored the impact of the feast/famine (F/F) ratio on the succession of autotrophic ammonia oxidizing bacteria (AOB) and HN-AD bacteria in a halophilic aerobic granular sludge (HAGS) system. As the F/F ratio decreased from 1/9 to 1/15, the total inorganic nitrogen (TIN) removal performance significantly decreased. The proportion of heterotrophic bacteria was dropped from 79.0 % to 33 %. Accordingly, the relative abundance of Paracoccus decreased from 70.8 % to 25.4 %, and the copy number of the napA gene was reduced from 2.2 × 1010 copies/g HAGS to 8.1 × 109 copies/g HAGS. It found the F/F ratio regulated the population succession of autotrophic AOB and HN-AD bacteria, thereby providing a solution to achieve the enrichment of HN-AD bacteria in HAGS.

Static magnetic field increases aerobic nitrogen removal from hypersaline wastewater in activated sludge with coexistence of fungi and bacteria.

Fungi have been found to exist in activated sludge treating saline wastewater, but their role in removing pollution has been neglected. This study explored the aerobic removal of total inorganic nitrogen (TIN) from saline wastewater under static magnetic fields (SMFs) with several strengths. Compared to the control, the aerobic removal of TIN was significantly increased by 1.47 times in 50 mT SMF, due to the increased dissimilation nitrogen removal by fungi and bacteria. Under SMF, fungal nitrogen dissimilation removal was significantly increased by 3.65 times. The fungal population size decreased, and its community composition changed significantly under SMF. In contrast, bacterial community composition and population remained relatively stable. Under SMFs, heterotrophic nitrification - aerobic denitrification bacteria Paracoccus and the fungi denitrifying Candida formed a synergistic interaction. This study elucidates the fungal role in aerobic TIN removal and provides an efficient solution to improve TIN removal from saline wastewater by SMF.

A halophilic aerobic-heterotrophic strain Halomonas venusta SND-01: Nitrogen removal by ammonium assimilation and heterotrophic nitrification-aerobic denitrification.

Nitrogen (N) removal from high-salinity wastewater is a major challenge. The aerobic-heterotrophic nitrogen removal (AHNR) process has been demonstrated to be feasible for treating hypersaline wastewater. In this study, Halomonas venusta SND-01, a halophilic strain capable of performing AHNR, was isolated from saltern sediment. The strain achieved ammonium, nitrite, and nitrate removal efficiencies of 98%, 81%, and 100%, respectively. The N balance experiment suggests that this isolate removes N mainly via assimilation. Various functional genes related to N metabolism were found in the genome of the strain, establishing a complex AHNR pathway that includes ammonium assimilation, heterotrophic nitrification-aerobic denitrification, and assimilatory nitrate reduction. Four key enzymes in the N removal process were successfully expressed. The strain exhibited high-adaptability under C/N ratios of 5-15, salinities of 2%-10% (m/v), and pH of 6.5-9.5. Therefore, the strain shows high potential for treating saline wastewater with different inorganic N compositions.

Spontaneous granulation of moderately halophilic sludge inoculated with saltern sediments from single granule into multi-granule aggregation.

There is very limited research on the application of moderate halophiles for biotreatment of hypersaline wastewater widely generated from some industries. This study demonstrated the development of moderate halophiles inoculated from saltern sediments into aerobic granule sludge (AGS) to treat hypersaline wastewater with a salinity of 100 g/L. The granulation of moderate halophiles can occur without applying the settling velocity selective pressure. The saltern sediment initially aggregated into single small granules and finally developed into 1200 ± 50 µm multiparticle granules. The halophiles affiliated in Halomonas was dominant in the granular bacterial community, with a relative abundance of 94.52%. Halomonas ventosae secreted sulfated polysaccharides. The sulfated polysaccharides content accounted for 63.95 ± 2.10% in the polysaccharides (PS), having an adhesive role in connecting single granules. Multiparticle granules showed the clear stratified structure, with α-D-glucopyranose polysaccharides in the inner bounders and ß-D-glucopyranose polysaccharides in the outer. The moderately granular sludge showed the stable chemical oxygen demand (COD) removal efficiency of >90% and the aerobic total inorganic nitrogen (TIN) removal efficiency (equal to ammonia removal) of 70 ± 5.00%. This paper contributes new insight into the formation of moderately halophilic granular sludge and accelerates the application of moderately halophilic granular sludge to treat hypersaline wastewater.

A novel Pseudomonas aeruginosa strain performs simultaneous heterotrophic nitrification-aerobic denitrification and aerobic phosphate removal.

Nitrogen and phosphate removal from wastewater relies on different functional bacteria. In this study, a novel strain affiliated with Pseudomonas aeruginosa was isolated from activated sludge by gradient dilution and performed heterotrophic nitrification-aerobic denitrification and aerobic phosphate removal (HNADPR). The strain showed an ammonium removal efficiency of 87% and a phosphate removal efficiency of 97% under optimal conditions, such as C/N ratio of 10, P/N ratio of 0.1, temperature of 30°C, and pH of 7.5-8.5. The modified Gompertz model could fit well the heterotrophic ammonium nitrification, aerobic nitrite/nitrate denitrification, and aerobic phosphate removal processes. Functional gene amplification indicated that ammonium removal followed the complete HN-AD pathway (NH4+ â†’ NH2OH â†’ NO2- â†’ NO3- â†’ NO2- â†’ NO â†’ N2O â†’ N2). Phosphate removal only occurred under aerobic conditions and ceased under anaerobic conditions. In successive aerobic cycles, the strain persistently took up phosphate. In wastewater, phosphate was aerobically converted into cell membrane, intracellular and extracellular polymeric substrates (EPS). Phosphorus in the form of phosphate monoester was pooled in EPS. A hypothetic aerobic phosphate removal model for strain SNDPR-01 is proposed to improve our understanding of the novel bacterial function of HNADPR.

Coupling Magnetic Field and Salinity Upshock To Improve Polyhydroxyalkanoate Productivity by Haloferax mediterranei Feeding on Molasses Wastewater.

Low polyhydroxyalkanoate (PHA) volumetric productivity from wastewater limits low-cost PHA production. To resolve this problem, an external magnetic field (MF) coupled with upshock salinity was applied to PHA production by Haloferax mediterranei (family Halobacteriaceae). Elevating the fermentation salinity over the optimal growth salinity (200 g/L) increased the PHA cell content while inhibiting cell proliferation, decreasing volumetric productivity. When a MF of 50 mT in 300 g/L salinity was applied, H. mediterranei proliferation and PHA cell content were promoted, leading to a 7.95% increase in PHA volumetric productivity in synthetic molasses wastewater and a 13.82% increase in glucose feeding compared with those in 200 g/L salinity. Under the MF, osmotic pressure regulation was activated by accumulating K+ and increasing betaine synthesis. The maximum betaine content increased by 74.33% in 300 g/L salinity with a 50-mT MF compared with that in 200 g/L salinity. When a 50-mT MF in 300 g/L salinity was applied, the malondialdehyde (MDA) content decreased by 32.66% and the activity of superoxide dismutase (SOD) increased by 46.89%, which reduced the oxidative damage. This study provides a new solution to enhance PHA volumetric productivity by MF and an insight into the magnetic effects of H. mediterranei. IMPORTANCE The obstacle to replacing petroplastics with PHA is its high production cost. To increase the fermentation economy, a novel strategy of coupling a MF with salinity upshock was applied, which enhanced the PHA volumetric productivity of H. mediterranei in fermenting molasses wastewater. The magnetic effect of H. mediterranei was found at a MF of 50 mT, which improved the salt tolerance of H. mediterranei and reduced the oxidative damage induced by the elevated salinity, thereby promoting proliferation and PHA cell content. This is the first time a technical method for enhancing PHA volumetric productivity by means of a MF has been proposed. Such a strategy can advance the utilization of H. mediterranei for the industrial production of PHA using organic wastewater.

Promoting enrichment of sulfur-oxidizing autotrophic denitrifiers via static magnetic fields: Performance and mechanism of magnetic biological effects.

Sulfur-driven autotrophic denitrification (SADN) is a promising technology for nitrogen removal from wastewater. In this study, different-strength SMFs (0, 5, 20, 50, 70 mT) were evaluated to investigate the potential of external static magnetic field (SMF) for enriching sulfur-oxidizing autotrophic denitrifiers (SOAD). 50-mT and 70-mT SMFs were most suitable to accelerate the growth of SOAD and the elimination of non-SOAD. The relative abundance of Thiobacillus significantly increased (p < 0.01) from 6.26% in control reactor to 36.15% under 50 mT and 52.51% under 70 mT. Under 50 mT, Thiobacillus denitrificans accumulated most rapidly, with the largest population. Furthermore, functional gene forecast by high-throughput and metagenomic sequencing indicated that SMF changed the two-component system, the adenosine 5'-monophosphate-activated protein kinase (AMPK) signaling system, the phosphotransferase system (PTS), as well as N/S-related enzymes to regulate stress-response and promote the growth of SOAD. The findings indicated that SMF accelerated the start-up of SADN.

Carbon Source Applied in Enrichment Stage of Mixed Microbial Cultures Limits the Substrate Adaptability for PHA Fermentation Using the Renewable Carbon.

Suitability of different substrates for enriched mixed microbial cultures (MMCs) is of importance to the polyhydroxyalkanoate (PHA) fermentation using renewable carbon. In this study, three enriched MMCs were evaluated for their fermentation features and kinetics with different carbon sources (sodium acetate, glucose, or starch). The results showed that the highly specific bacterial community composition was developed depending on the applied carbon source. Correspondence analysis suggested that the genus affiliated in Gammaproteobacteria_unclassified was related to 3-hydroxybutyrate (HB) synthesis in acetate-fed MMC (relative abundance of 38%) and glucose-fed MMC (relative abundance of 76.7%), whereas Vibrio genus was related to 3-hydroxyvalerate (HV) production in glucose-fed MMC (relative abundance of 0.4%) and starch-fed MMC (relative abundance of 94.6%). The acetate-fed MMC could not use glucose and starch as fermentation carbon sources, showing the limitation of microbial species developed with the specific metabolic substrate. Glucose-fed MMC produced the highest PHA cell content of 64.2% cell dry weight when using sodium acetate as the fermentation carbon. Glucose-fed MMC showed wide resilience and adaptation to various carbon sources. When actual landfill leachate was used for fermentation by glucose-fed MMC, maximum PHA cell content of 45.5% cell dry weight and the PHA volumetric productivity of 0.265 g PHA/(L·h) were obtained. This study suggested carbon sources applied in the MMC enrichment stage had a significant influence on utilization of carbon in the fermentation stage.

A novel halophilic Exiguobacterium mexicanum strain removes nitrogen from saline wastewater via heterotrophic nitrification and aerobic denitrification.

The utilization of halophilic bioresources is limited due to a lack of isolation and characterization work. A halophilic bacterium strain SND-01 of Exiguobacterium mexicanum was isolated in this study, which is the first report on its novel function in heterotrophic nitrification-aerobic denitrification (HN-AD). The strain SND-01 is slightly halophilic, surviving at 0 up to 9% (w/v) salinity. When utilizing ammonium, nitrate or nitrite as the sole nitrogen source in aerobic conditions, the isolated strain showed the maximum nitrogen removal rate of 2.24 ± 0.14 mg/(L·h), 3.63 ± 0.21 mg/(L·h) and 2.30 ± 0.23 mg/(L·h), respectively. Functional genes and key enzymes involved in heterotrophic-aerobic nitrogen transformations were characterized, establishing the pathway of HN-AD. The nitrogen removal via HN-AD is dependent on the C/N ratio, salinity and temperature. The halophilic Exiguobacterium mexicanum strain SND-01 shows a significant potential in biotreatment of saline wastewater in an easy and cost-effective way.

The metabolic pathways of polyhydroxyalkanoates and exopolysaccharides synthesized by Haloferax mediterranei in response to elevated salinity.

How polymer synthesis is mobilized or activated as a biological response of Haloferax mediterranei against hypertonic conditions remains largely unexplored. This study investigated the protein expression of H. mediterranei in response to high salinity by using isobaric tags for relative and absolute quantitation (iTRAQ)-based proteomic analysis. The microbes were harvested at end of fermentation at the NaCl salinity of 75 and 250 g L-1. Among the identified 2123 proteins, 170 proteins were differentially expressed. Gene ontology annotation revealed that the highest number of proteins was annotated in biological process category, which was responsible for metabolic process, cellular component and catalytic activity. Differentially expressed proteins were belonged to the class of response to stimulus as well as catalytic activity and binding. Under high salinity conditions, three pathways were established as key responses of PHA and EPS production to hypertonic pressure. Two overexpressed proteins, beta-ketoacyl-ACP reductase and 3-hydroxyacyl-CoA dehydrogenase, enhanced the synthesis of PHAs. The serine-pyruvate transaminase and serine-glyoxylate transaminase were upregulated, thereby increasing the conversion of glucose to PHA. Downregulated levels of sulfate-adenylyl transferase and adenylyl-sulfate kinase could cause diminished EPS synthesis. This study could contribute to better understanding of the proteomic mechanisms of the synthesized polymers in defending against salt stress. SIGNIFICANCE: Haloferax mediterranei, a family member of halophilic archaea, is well known for its fermentative production of poly-ß-hydroxyalkanoates (PHAs). PHAs are natural polymers that exhibit great potential in a wide range of applications such as a good alternative to petroleum-based plastics and the biocompatible material. For decades, the functional role of PHAs synthesized by H. mediterranei is deemed to be carbon and energy reservations. The finding proved that differential production of PHA and EPS in H. mediterranei exposed to elevated salinity was caused by differential protein expression. This is the first report on how PHA and EPS synthesized by H. mediterranei is mobilized as the response of increased salinity, contributing to the understanding of halophilic archaea's response to hypertonic stress and the precise control of fermentation production. Despite its advantages as a PHA cell factory, H. mediterranei synthesized EPS simultaneously, thereby lowering the maximum yield of PHA production. Overall, salinity can be used as a vital microbial fermentation parameter to obtain the highest harvest of PHA, as well as the lowest EPS synthesis in industrial fermentation.

Fast granulation of halophilic activated sludge treating low-strength organic saline wastewater via addition of divalent cations.

Granulation of halophilic activated sludge is an important solution to solve the problem of solid-liquid separation in biological treatment of saline wastewater. This study demonstrated that by adding divalent cations into the saline influent with low organic load, halophilic granular sludge with an average diameter of 910 ± 10 µm can be cultivated. The close correlation between divalent cations and particle size indicated that Ca2+ played a major role in the granulation process. Ca2+ was accumulated in halophilic granular sludge, which provided an inorganic carrier for microbial aggregation and leaded to the dominance of halophilic bacteria of the family Flavobacteriaceae. The halophilic bacteria secreted a large amount of extracellular polymeric substances (EPS), which contained 70.0 ± 0.02% protein. By enhancing the EPS network of protein and Ca2+, halophilic granular sludge was formed. The addition of Mg2+ enhanced the network of Mg2+ and loosely bound EPS, which could be destroyed due to Na+ substitution. This study provides an effective granulation method for halophilic activated sludge.

A laser-induced breakdown spectroscopy-integrated lateral flow strip (LIBS-LFS) sensor for rapid detection of pathogen.

Incorporation of new readout methods with established analytical devices allows methodological innovations in analytical sciences. Herein, we present a new sensing platform by combining an ultrasensitive element analyzer, namely the laser-induced breakdown spectroscopy (LIBS) and a lateral flow strip (LFS). AgxAuy bimetallic nanoparticles (AgxAuyBNPs) are selected as the labels to deliver the optimal quantitative performance by analyzing the Ag (I) signal from the test (T) line of LFS. For prototypical application in pathogen detection, the LIBS-LFS sensor can achieve a detection limit of 1.6 cfu mL-1 of Staphylococcus aureus (S. aureus) within 10 min, which is superior to conventional methods. Importantly, the signals of AgxAuyBNPs for visual and LIBS analysis are stable and still readable after the detection is finished and the test strip is stored for up to 13 days, suggesting a potential for long-term data preservation. This combination of LIBS with LFS provides a new concept toward integrated nano/analytical devices that can benefit various application scenarios.

Granulation of halophilic sludge inoculated with estuarine sediments for saline wastewater treatment.

As a solution of the sludge loss in the treatment of saline wastewater, the granulation of halophilic sludge was explored in this study. The inoculated estuarine sediment was granulated to an average diameter of 1155 ±â€¯102 µm under the selective settling pressure in the airlift sequencing batch reactor (SBR) when the influent organic loading rate (OLR) was doubled to 0.36 g COD/L·day. The results indicated that the OLR doubled the amount of total extracellular polymeric substance (EPS) and that protein was predominant in the EPS (72.8 ±â€¯2.0%). The correlation between aggregate size and protein content was better than that between aggregate size and polysaccharide content. The amount of alginate-like exopolysaccharides (ALE) increased linearly at the mature granular stage, co-occurring with the compact and elastic structure of the granules. According to the results of 16S rRNA high -throughput sequencing, the Shannon-Weaver index of mature granule decreased by >50% compared to the inoculated sediment. Bacteria of Propionibacteriaceae family constituted 34% of the population in granules and were in symbiotic relationship with halophiles of family Rhodocyclaceae, Vibrionaceae, Flavobacteriaceae, and Cryomorphaceae. The aerobic halophilic granular sludge showed COD removal efficiency of 90.9 ±â€¯0.8% and ammonia removal efficiency of 72.6 ±â€¯4.0% for 30 g/L saline wastewater. An average nitrite accumulation ratio of 94.5 ±â€¯2.9% was observed during nitrification. Granulation of halophilic sludge provides an effective solution to the saline sludge loss problem, which is a step forward to realize the biological treatment of saline wastewater by halophiles.

[Effect of Carbon Sources on the Accumulation of Endogenous Polymers and Denitritation in the O/A-F/F Mode].

To accumulate endogenous polymers during the aerobic phase, the aerobic/anoxic-feast/famine (O/A-F/F) selection mode can be used. It can also be used in situ for endogenous denitrification by activated sludge during the anoxic phase. To further explore the effect of carbon sources on the activated sludge accumulation of endogenous polymers and endogenous denitrification, this study used acetic and glucose as the main carbon sources to investigate the accumulation of endogenous polymers, endogenous denitrification, and the structure and function of enriched activated sludge. The results show that acetic (Ac-SBR) and glucose (Gc-SBR) as the main carbon source systems achieved a 40 mg·L-1 nitrate removal by endogenous denitrification when the influent chemical oxygen demand (COD) was~500 mg·L-1 in the O/A-F/F selection mode. Both the Ac-SBR and Gc-SBR achieved partial denitrification, but the nitrite accumulation of the Ac-SBR was higher than that of the Gc-SBR. Acetic is favorable for the accumulation of endogenous polyhydroxyalkanoate (PHA); PHA drives the endogenous denitrification. The yield of PHA was 0.52 and the denitrification rate (DNR) was 9.65 mg·(L·h)-1. The Gc-SBR system achieved the simultaneous accumulation of PHA and glycogen (Gly). The yield of Gly was higher than that of PHA and the DNR driven by Gly was 4.35 mg·(L·h)-1. The Gly was the main driving force to achieve endogenous denitrification and contributed to 77% of the total nitrogen removal. The 16S rRNA high-throughput sequencing analysis of activated sludge flora shows that the class of ß-Proteobacteria in the Proteobacteria was dominant, with an abundance of 40.56% in the Ac-SBR. However, the abundance of ß-Proteobacteria was only 18.05% in the Gc-SBR. The class of α-Proteobacteria contributes to glycogen accumulation in the Gc-SBR. The PHA can be accumulated by ß-Proteobacteria, Unclassified Bacteroidetes, and Lgnavibacteria in the Ac-SBR.

Cr(VI) Adsorption on Red Mud Modified by Lanthanum: Performance, Kinetics and Mechanisms.

Water pollution caused by the highly toxic metal hexavalent chromium (Cr(VI)) creates significant human health and ecological risks. In this study, a novel adsorbent was used to treat Cr(VI)-containing wastewater; the adsorbent was prepared using red mud (RM) generated from the alumina production industry and the rare earth element lanthanum. This study explored adsorption performance, kinetics, and mechanisms. Results showed that the adsorption kinetics of the RM modified by lanthanum (La-RM), followed the pseudo-second-order model, with a rapid adsorption rate. Cr(VI) adsorption was positively associated with the absorbent dose, pH, temperature, and initial Cr(VI) concentration; coexisting anions had little impact. The maximum Cr(VI) adsorption capacity was 17.35 mg/g. Cr(VI) adsorption on La-RM was a mono-layer adsorption pattern, following the Langmuir isotherm model. Thermodynamic parameters showed the adsorption was spontaneous and endothermic. The adsorption of Cr(VI) on La-RM occurred as a result of LaOCl formation on the RM surface, which in turn further reacted with Cr(VI) in the wastewater. This study highlighted a method for converting industrial waste into a valuable material for wastewater treatment. The novel absorbent could be used as a potential adsorbent for treating Cr(VI)-contaminating wastewater, due to its cost-effectiveness and high adsorption capability.

Effects of carbon sources on the enrichment of halophilic polyhydroxyalkanoate-storing mixed microbial culture in an aerobic dynamic feeding process.

Microbial polyhydroxyalkanoate (PHA) production serves as a substitute for petroleum-based plastics. Enriching mixed microbial cultures (MMCs) with the capacity to store PHA is a key precursor for low-cost PHA production. This study investigated the impact of carbon types on enrichment outcomes. Three MMCs were separately fed by acetate sodium, glucose, and starch as an enriching carbon source, and were exposed to long-term aerobic dynamic feeding (ADF) periods. The PHA production capacity, kinetics and stoichiometry of the enrichments, the PHA composition, and the microbial diversity and community composition were explored to determine carbon and enrichment correlations. After 350-cycle enriching periods under feast-famine (F-F) regimes, the MMCs enriched by acetate sodium and glucose contained a maximum PHA content of 64.7% and 60.5% cell dry weight (CDW). The starch-enriched MMC only had 27.3% CDW of PHA. High-throughput sequencing revealed that non-PHA bacteria survived alongside PHA storing bacteria, even under severe F-F selective pressure. Genus of Pseudomonas and Stappia were the possible PHA accumulating bacteria in acetate-enriched MMC. Genus of Oceanicella, Piscicoccus and Vibrio were found as PHA accumulating bacteria in glucose-enriched MMC. Vibrio genus was the only PHA accumulating bacteria in starch-enriched MMC. The community diversity and composition were regulated by the substrate types.