Erik R. Coats

Polyhydroxybutyrate synthesis on biodiesel wastewater using mixed microbial consortia.

Crude glycerol (CG), a by-product of biodiesel production, is an organic carbon-rich substrate with potential as feedstock for polyhydroxyalkanoate (PHA) production. PHA is a biodegradable thermoplastic synthesized by microorganisms as an intracellular granule. In this study we investigated PHA production on CG using mixed microbial consortia (MMC) and determined that the enriched MMC produced exclusively polyhydroxybutyrate (PHB) utilizing the methanol fraction. PHB synthesis appeared to be stimulated by a macronutrient deficiency. Intracellular concentrations remained relatively constant over an operational cycle, with microbial growth occurring concurrent with polymer synthesis. PHB average molecular weights ranged from 200-380 kDa, while thermal properties compared well with commercial PHB. The resulting PHB material properties and characteristics would be suitable for many commercial uses. Considering full-scale process application, it was estimated that a 38 million L (10 million gallon) per year biodiesel operation could potentially produce up to 19 metric ton (20.9t on) of PHB per year.

Post-anoxic denitrification driven by PHA and glycogen within enhanced biological phosphorus removal.

The objective of this research was to interrogate and develop a better understanding for a process to achieve post-anoxic denitrification without exogenous carbon augmentation within enhanced biological phosphorus removal (EBPR). Sequencing batch reactors fed real wastewater and seeded with mixed microbial consortia were operated under variable anaerobic-aerobic-anoxic and organic carbon loading conditions. The process consistently achieved phosphorus and nitrogen removal, while the observed specific denitrification rates were markedly higher than expected for post-anoxic systems operated without exogenous organic carbon addition. Investigations revealed that post-anoxic denitrification was predominantly driven by glycogen, an intracellular carbon storage polymer associated with EBPR; moreover, glycogen reserves can be significantly depleted post-anoxically without compromising EBPR. Success of the proposed process is predicated on providing sufficient organic acids in the influent wastewater, such that residual nitrate carried over from the post-anoxic period is reduced and polyhydroxyalkanoate (PHA) synthesis occurs.

Lactic acid production with undefined mixed culture fermentation of potato peel waste

Potato peel waste (PPW) as zero value byproduct generated from food processing plant contains a large quantity of starch, non-starch polysaccharide, lignin, protein, and lipid. PPW as one promising carbon source can be managed and utilized to value added bioproducts through a simple fermentation process using undefined mixed cultures inoculated from wastewater treatment plant sludge. A series of non-pH controlled batch fermentations under different conditions such as pretreatment process, enzymatic hydrolysis, temperature, and solids loading were studied. Lactic acid (LA) was the major product, followed by acetic acid (AA) and ethanol under fermentation conditions without the presence of added hydrolytic enzymes. The maximum yields of LA, AA, and ethanol were respectively, 0.22 g g(-1), 0.06 g g(-1), and 0.05 g g(-1). The highest LA concentration of 14.7 g L(-1) was obtained from a bioreactor with initial solids loading of 60 g L(-1) at 35°C.

Advancing post-anoxic denitrification for biological nutrient removal

The objective of this research was to advance a fundamental understanding of a unique post-anoxic denitrification process for achieving biological nutrient removal (BNR), with an emphasis on elucidating the impacts of surface oxygen transfer (SOT), variable process loadings, and bioreactor operational conditions on nitrogen and phosphorus removal. Two sequencing batch reactors (SBRs) were operated in an anaerobic/aerobic/anoxic mode for over 250 days and fed real municipal wastewater. One SBR was operated with a headspace open to the atmosphere, while the other had a covered liquid surface to prevent surface oxygen transfer. Process performance was assessed for mixed volatile fatty acid (VFA) and acetate-dominated substrate, as well as daily/seasonal variance in influent phosphorus and ammonia loadings. Results demonstrated that post-anoxic BNR can achieve near-complete (>99%) inorganic nitrogen and phosphorus removal, with soluble effluent concentrations less than 1.0 mgN L(-1) and 0.14 mgP L(-1). Observed specific denitrification rates were in excess of typical endogenous values and exhibited a linear dependence on the glycogen concentration in the biomass. Preventing SOT improved nitrogen removal but had little impact on phosphorus removal under normal loading conditions. However, during periods of low influent ammonia, the covered reactor maintained phosphorus removal performance and showed a greater relative abundance of polyphosphate accumulating organisms (PAOs) as evidenced by quantitative real-time PCR (qPCR). While GAOs were detected in both reactors under all operational conditions, BNR performance was not adversely impacted. Finally, secondary phosphorus release during the post-anoxic period was minimal and only occurred if nitrate/nitrite were depleted post-anoxically.

Functional stability of a mixed microbial consortium producing PHA from waste carbon sources

Polyhydroxyalkanoates (PHAs) represent an environmentally effective alternative to synthetic thermoplastics; however, current production practices are not sustainable. In this study, PHA production was accomplished in sequencing batch bioreactors utilizing real wastewaters and mixed microbial consortia from municipal activated sludge as inoculum. Polymer production reached 85, 53, and 10% of the cell dry weight from methanol-enriched pulp and paper mill foul condensate, fermented municipal primary solids, and biodiesel wastewater, respectively. Using denaturing gradient gel electrophoresis of 16S-rDNA from polymerase chain reaction-amplified DNA extracts, distinctly different communities were observed between and within wastewaters following enrichment. Most importantly, functional stability was maintained despite differing and contrasting microbial populations.

Toward Polyhydroxyalkanoate Production Concurrent with Municipal Wastewater Treatment in a Sequencing Batch Reactor System

Bacteria can synthesize cytoplasmic granules known as polyhydroxyalkanoates PHAs, which are carbon and energy storage reserves, from organic carbon when subject to stressful environmental conditions. PHAs are also biodegradable thermoplastics with many potential commercial applications. The purpose of the research reported herein was to evaluate the feasibility of integrating PHA production within a municipal wastewater treatment WWT configured as a sequencing batch reactor SBR. Four bench-scale WWT SBRs were tested at decreasing organic loading rates to assess the potential to enrich for microbes capable of feast/famine PHA synthesis. For each treatment SBR, sidestream batch reactors receiving higher quantities of primary solids fermenter liquor were operated to produce PHA. Results from this study demonstrate that a treatment SBR supplied moderate strength wastewater can enrich for the target microorganisms, with PHA yields of 0.23-0.31-mg PHA per mg chemical oxygen demand, and produce high quality effluent. In side- stream batch reactors, microorganisms that fed excess quantities of substrate can rapidly synthesize significant quantities of PHA. Based on the results of this study, we estimate that a 1 million gallon per day SBR WWT-PHA production system could generate 11-36 t 12-40 t of PHA annually.

Comparative analysis of microbial community of novel lactic acid fermentation inoculated with different undefined mixed cultures

Three undefined mixed cultures (activated sludge) from different municipal wastewater treatment plants were used as seeds in a novel lactic acid fermentation process fed with potato peel waste (PPW). Anaerobic sequencing batch fermenters were run under identical conditions to produce predominantly lactic acid. Illumina sequencing was used to examine the 16S rRNA genes of bacteria in the three seeds and fermenters. Results showed that the structure of microbial communities of three seeds were different. All three fermentation products had unique community structures that were dominated (>96%) by species of the genus Lactobacillus, while members of this genus constituted <0.1% in seeds. The species of Lactobacillus sp. differed among the three fermentations. Results of this study suggest the structure of microbial communities in lactic acid fermentation of PPW with undefined mixed cultures were robust and resilient, which provided engineering prospects for the microbial utilization of carbohydrate wastes to produce lactic acid.

Effect of organic loading and retention time on dairy manure fermentation

The investigations presented and discussed herein establish an enhanced understanding on volatile fatty acid (VFA) production as a function of dairy manure fermenter organic loading (OL) and retention time (RT), first through a factorial of 64 fermentation potential (FP) batch tests, followed by analysis of a continuously operated pilot-scale fermenter. The maximum observed net FP - 0.103 mg VFA produced (as COD) (mg VS applied)(-1) - occurred at an OL of 40.7 g VS L(-1) and at a RT of 6 days. The pilot-scale fermenter exhibited an average yield of 0.09 mg VFA (as COD) synthesized (mg VS applied)(-1), with average effluent total VFA concentrations of 6398 mg VFA (as COD) L(-1). The research demonstrates that FP tests are an effective method to optimize continuously operated dairy manure fermenters, and that dairy manure fermentation can yield large quantities of organic acids at short RTs and high OL rates.

Lactic acid production from potato peel waste by anaerobic sequencing batch fermentation using undefined mixed culture

Lactic acid (LA) is a necessary industrial feedstock for producing the bioplastic, polylactic acid (PLA), which is currently produced by pure culture fermentation of food carbohydrates. This work presents an alternative to produce LA from potato peel waste (PPW) by anaerobic fermentation in a sequencing batch reactor (SBR) inoculated with undefined mixed culture from a municipal wastewater treatment plant. A statistical design of experiments approach was employed using set of 0.8L SBRs using gelatinized PPW at a solids content range from 30 to 50 g L(-1), solids retention time of 2-4 days for yield and productivity optimization. The maximum LA production yield of 0.25 g g(-1) PPW and highest productivity of 125 mg g(-1) d(-1) were achieved. A scale-up SBR trial using neat gelatinized PPW (at 80 g L(-1) solids content) at the 3 L scale was employed and the highest LA yield of 0.14 g g(-1) PPW and a productivity of 138 mg g(-1) d(-1) were achieved with a 1 d SRT.

Influence of organic loading rate and solid retention time on polyhydroxybutyrate production from hybrid poplar hydrolysates using mixed microbial cultures.

The aim of this study was to investigate the potential of using wood hydrolysates (enzymatically hydrolyzed from hybrid poplar) as substrate to produce polyhydroxybutyrate (PHB) using mixed microbial cultures. The optimal operational conditions for fed-batch bioreactors were 4d solid retention time with an organic loading rate of 2.5g/Ld. The maximum PHB accumulated was 27% of cell dry weight with a yield of 0.32g/g (g PHB produced per g sugars consumed). Microbial community analysis was done at the genus level by 16S rRNA sequencing on an Illumina system and community evolution was observed among different samples and initial seed. Actinobacteria, Alpha- and Beta-proteobacteria were found to be the dominant groups in all the bioreactors. Several PHB-storing microorganisms were characterized belonging to Alpha- and Beta-proteobacteria.