Laboratory-scale Bioreactor Research Articles

Self-Forming Dynamic Membrane BioReactors (SFD MBR) for municipal wastewater treatment: Relevance of solids retention time and biological process stability

A lab-scale Self-Forming Dynamic Membrane BioReactor (SFD MBR) equipped with a nylon mesh was operated for the treatment of real municipal wastewater under a volumetric loading rate of 0.9–1.1 gCOD L−1 d−1 and a flux of 61–68 L m−2 h−1. During an experimental period of more than 600 d, three runs were performed with different solids retention times (SRT of 15, 30, and 50 d). At the steady state, the system had always good biological and filtration performances, but the mesh clogging tendency and the solid removal observed varied with the SRT. Lower filtration performance, both in terms of permeate quality and productivity, was observed under unstable conditions for the biological processes, such as start-up and variations of the food-to-microorganism ratio. Higher SRT resulted in more stable biological process at the steady state, which positively affected the filtration performance even in front of organic load variations.

High-yield production of active recombinant S. simulans lysostaphin expressed in E. coli in a laboratory bioreactor

Staphylococcus aureus (S. aureus), which has developed multidrug resistance, leads to many healthcare-associated infections resulting in significant medical and economic losses. Therefore, the development of new efficient strategies to deal with these bacteria has been gaining importance. Lysostaphin is a peptidoglycan hydrolase that has considerable potential as a bacteriocin. However, there have been few reported optimization and scale-up studies of the lysostaphin bioproduction process. Our preliminary results have revealed that the composition of auto-induction media at 30 °C increases the produced lysostaphin around 10-fold in shake flasks. In this study, achieving higher yields for recombinant lysostaphin in E. coli at a laboratory scale has been the aim, through the use of auto-induction media.Optimized medium composition and fermentation parameters were transferred to a laboratory-scale bioreactor. The tested conditions improved protein yields up to 184 mg/L in a 3 L stirred bioreactor and the productivity was improved 2-fold in comparison to previously published reports. Furthermore, this study also showed that lysostaphin is an effective bacteriocin on both commercially available and isolated S. aureus strains. These results will contribute to future larger-scale production of lysostaphin via the proposed fermentation conditions.

Metagenomic Study of Bacterial and Archaeal Populations during Anaerobic Digestion of Lignocellulosic Waste in Lab-scale Biogas Reactors

This study evaluated using 16S rDNA gene-based metagenomics technique the populations of bacteria and archaea in digestate samples from lab-scale anaerobic bioreactors digesting pretreated and untreated coconut husk fiber, pineapple floret and banana stem. Result of biodegradability experiment indicated high microbial activity in digestate (biogas slurry), with untreated banana stem having the highest total solids (TS) and volatile solids (VS) removal efficiencies of 78.3 % and 92.9 % respectively. Similarly, all pretreated substrates exhibited higher TS and VS losses with corresponding TS (77.8 %) and VS (87.2 %) removal efficiencies. This TS and VS removal rates signaled increased rate of organic matter decomposition with concomitant biogas productivity. Diversity comparisons performed between samples showed rich microbial diversity in untreated sample than the pretreated sample. Taxonomic composition revealed that, for untreated samples at the phylum level, the bacterial community was predominantly Firmicutes (relative abundance 97.0 %), with 0.30 % Actinobacteria and 0.10 % Proteobacteria. The genus Oxobacter (35.0 %), Clostridium (12.0 %) and Ethanoligenens (10.0 %) were ubiquitous and abundant in the untreated sample. The archaeal community was however dominated by the Euryarchaeota with one methanogenic order Methanomicrobiales, and a high abundance of the genera Thermacetogenium. For pretreated samples, at the phylum level, bacterial community was also dominated by Firmicutes (95.0 %), followed by Proteobacteria (1.02 %), Actinobacteria (0.18 %) and Tenericutes (0.06 %). The genus Clostridium (41.0 %), Ethanoligenens (29.0 %) and Lactobacillus (15.0 %) were also ubiquitous and abundant in the pretreated sample. Archaeal community was also dominated by Euryarchaeota with the two methanogenic orders Methanomicrobiales and Methanosarcinales dominating. The major microbial groups were hydrolyzing and fermenting populations. These findings revealed rich microbial assemblage and diversity among microbial communities in biogas digestate.

Influence of Hydraulic Retention Time (HRT) upon the Treatment of Wastewater by a Laboratory-Scale Membrane Bioreactor (MBR)

The effect of applying different hydraulic retention times (HRTs) was investigated for a lab-scale membrane bioreactor (MBR) treating domestic wastewater. The submerged flat-type ultrafiltration MBR was operated under three HRTs (576 min, 462 min, and 372 min) corresponding to three operating periods to investigate its efficiency in removing organic content and nutrients from domestic wastewater. The chemical oxygen demand (COD) decreased from 99 to 96.5% with the HRT decreased from 576 min to 462 min. The bacteria performing nitrification were affected by the HRT change: the ammonium (NH4-N) removal dropped from 98.5% (HRT of 576 min) to 74.2% (HRT of 462 min). With the HRT adjusted to the lowest value of 372 min, the COD and NH4-N removals were 94.4% and 55.8%, respectively. The phosphate (PO4-P) removal was 59.1%, 34.4%, and 28% during periods 1, 2 and 3, respectively. In terms of the COD removal efficiency, the treated effluent met the Turkish limits for discharge to the receiving water during all of the operating periods. The system performance was suitable in terms of NH4-N removal for periods 1 (HRT of 576 min) and 2 (HRT of 462 min). However, applying the operating conditions of period 2 (i.e., maintaining the HRT at 462 min) requires additional post-treatment (e.g., low-cost chemical precipitation) to enhance the PO4-P removal.

Autotrophic Fixed-Film Systems Treating High Strength Ammonia Wastewater.

The aim of the study was enrichment of nitrifying bacteria and to investigate the potential of autotrophic fixed-film and hybrid bioreactors to treat high strength ammonia wastewater (up to 1,000 mg N/L). Two types of fixed-film systems [moving bed biofilm reactor (MBBR) and BioCordTM] in two different configurations [sequencing batch reactor (SBR) and a continuous stirred tank reactor (CSTR)] were operated for 306 days. The laboratory-scale bioreactors were seeded with activated sludge from a municipal wastewater treatment plant and fed synthetic wastewater with no organics. Strategies for acclimation included biomass reseeding (during bioreactor start-up), and gradual increase in the influent ammonia concentration [from 130 to 1,000 mg N/L (10% every 5 days)]. Stable ammonia removal was observed up to 750 mg N/L from 45 to 145 days in the MBBR SBR (94–100%) and CSTR (72–100%), and BioCordTM SBR (96–100%) and CSTR (92–100%). Ammonia removal declined to 87% ± 6, in all bioreactors treating 1,000 mg N/L (on day 185). Following long-term operation at 1,000 mg N/L (on day 306), ammonia removal was 93–94% in both the MBBR SBR and BioCordTM CSTR; whereas, ammonia removal was relatively lower in MBBR CSTR (20–35%) and BioCordTM SBR (45–54%). Acclimation to increasing concentrations of ammonia led to the enrichment of nitrifying (Nitrosomonas, Nitrospira, and Nitrobacter) and denitrifying (Comamonas, OLB8, and Rhodanobacter) bacteria [16S rRNA gene sequencing (Illumina)] in all bioreactors. In the hybrid bioreactor, the nitrifying and denitrifying bacteria were relatively more abundant in flocs and biofilms, respectively. The presence of dead cells (in biofilms) suggests that in the absence of an organic substrate, endogenous decay is a likely contributor of nutrients for denitrifying bacteria. The nitrite accumulation and abundance of denitrifying bacteria indicate partial denitrification in fixed-film bioreactors operated under limited carbon conditions. Further studies are required to assess the contribution of organic material produced in autotrophic biofilms (by endogenous decay and soluble microbial products) to the overall treatment process. Furthermore, the possibility of sustaining autotrophic nitrogen in high strength waste-streams in the presence of organic substrates warrants further investigation.

Powdered Ceramsite and Powdered Limestone Use in Aerobic Granular Sludge Technology.

The effects of two powdered mineral materials (powdered ceramsite and powdered limestone) on aerobic granulation of sludge were evaluated. The experiment was conducted on a laboratory scale bioreactors treating wastewater for 89 days. Three granular sequencing batch reactors (GSBRs) were operated at the lowest optimal organic loading rate (OLR) of 2.55 g COD/(L∙d). In the control reactor (R1), the mean diameter (d) of the biomass ranged from 124.0 to 210.0 µm, and complete granulation was not achieved. However, complete granulation did occur in reactors to which either ceramsite (251.9 µm < d < 783.1 µm) or limestone (246.0 µm < d < 518.9 µm) was added. Both powdered materials served as a ballast for the sludge flocs making up the seed sludge. Ceramsite particles also acted as microcarriers of granule-forming biomass. The granules in the reactors with added powdered materials had nonfibrous and smoother surfaces. The reactor with ceramsite exhibited the highest average efficiencies for COD, total nitrogen, and total phosphorus removal (85.4 ± 5.4%, 56.6 ± 10.2%, and 56.8 ± 9.9%, respectively). By contrast, the average nitrification efficiency was 95.1 ± 12.8%.

Decolourization of textile effluent using native microbial consortium enriched from textile industry effluent

A robust and efficient treatment process is required to address the problem of residual colour and avoid expensive post-treatment steps while dealing with textile effluents. In the present work, a novel microbial consortium enriched from textile effluent was used to optimize the process of decolourization under extreme conditions with minimum inputs. With PreTreatment Range (PTR) effluent as a carbon source and only 0.5 g/L yeast extract as external input, the process enabled 70-73% colour reduction (from 1910-1930 to 516-555 hazen) in dyeing unit wastewater. Unhindered performance at higher temperatures (30 °C-50 °C) and wide pH range (7-12) makes this process highly suitable for the treatment of warm and extremely alkaline textile effluents. No significant difference was observed in the decolourization efficiency for effluents from different batches (Colour: 1647-4307 hazen; pH-11.5-12.0) despite wide variation in nature and concentration of dyes employed. Long term (60 days) continuous mode performance monitoring at hydraulic retention time of 48 h in lab-scale bioreactor showed consistent colour (from 1734-1980 to 545-723 hazen) and chemical oxygen demand (1720-2170 to 669-844 mg/L) removal and consistently neutral pH of the treated water. Present study thus makes a significant contribution by uncovering the ability of native microbial consortium to reliably treat dye laden textile wastewater without any dilution or pre-treatment and with minimum external inputs. The results ensure easy applicability of this indigenously developed process at the industrial scale.

Xanthomonas campestris biocontrol agent: Selection, medium formulation and bioprocess kinetic analysis

Black rot, caused by Xanthomonas campestris pv. campestris, is one of the most important diseases of cruciferous crops which causes significant yield losses. Biological control of black rot by microbial biocontrol agents represents a promising alternative to chemical treatments and good agricultural practices which show only limited success. This study was carried out to assess a potential of different antagonists, including Bacillus, Pseudomonas, Lactobacillus, Streptomyces, Saccharomyces and Trichoderma genera, for biological control of black rot. Cultivation broth samples and their filtrates were examined against seven X. campestris strains, isolated from diseased cruciferous plants, using the diffusion-disc method. Bacillus velezensis has showed the highest inhibition zone diameter of 35.62?3.76 mm. Afterwards, different combinations of carbon and nitrogen sources were used in the cultivation medium to maximize antagonistic activity of B. velezensis. The best combinations were glycerol and yeast extract, lactose and peptone, as well as sucrose and yeast extract, suggesting the potential of biodiesel, dairy and sugar industry effluents in the production of bioactive compounds effective against the black rot pathogen. The validation experiment was performed in a laboratory-scale bioreactor, in order to investigate bioprocess kinetics of biomass growth and carbon source consumption, using the cultivation medium containing the optimal carbon and nitrogen source.

Effects of Solids Retention Time on the Anaerobic Membrane Bioreactor with Yttria-Based Ceramic Membrane Treating Domestic Wastewater at Ambient Temperature.

The effects of solid retention times (SRTs) (100 days, 50 days, 25 days) on the performance, microbial community, and membrane fouling of a lab-scale anaerobic yttria-based ceramic membrane bioreactor (AnCMBR) treating synthetic domestic wastewater at ambient temperature (31.2 ± 2.7 °C) were examined. The soluble chemical oxygen demand (SCOD) removal was higher (89.6%) at 25 days SRT compared with 50 days (39.61%) and 100 days (34.3%) SRT. At 100 days SRT, more Bacteroidetes, Firmicutes, and Proteobacteria were present in the microbial community. At 25 days SRT, more Chloroflexi, Synergistetes, and Pastescibacteria emerged, contributing to the stable performance. The SRT of 25 days has resulted in a more stable microbial community compared with 50 days and 100 days SRT. Both bacterial and archaeal community diversities were higher at 25 days SRT, and the specific production of soluble microbial by-products (SMPs) and extracellular polymeric substances (EPSs) were higher at 25 days SRT as well. Consequently, the membrane flux was lower at 25 days SRT with the increased particle size and the enhanced SMPs and EPSs production. Fourier transform infrared spectroscopy analysis (FTIR) and three-dimensional excitation and emission matrix (3D-EEM) analysis showed that protein and SMPs were the major membrane foulants at all SRT stages. In this study, SRT at 25 days was favorable for the stable operation of an AnCMBR treating domestic wastewater at ambient temperature.

Anaerobic membrane bioreactor performance at different wastewater pre-concentration factors: An experimental and economic study

This research evaluated the performance of a lab-scale anaerobic membrane bioreactor (AnMBR) treating municipal sewage pre-concentrated by forward osmosis (FO). The organic loading rate (OLR) and sodium concentrations of the synthetic sewage stepwise increased from 0.3 to 2.0 g COD L−1 d−1 and from 0.28 to 2.30 g Na+ L−1 to simulate pre-concentration factors of 1, 2, 5 and 10. No major operational problems were observed during AnMBR operation, with COD removal efficiencies ranging between 90 and 96%. The methane yield progressively increased from 214 ± 79 to 322 ± 60 mL CH4 g−1 COD as the pre-concentration factor increased from 1 to 10. This was mainly attributed to the lower fraction of methane dissolved lost in the permeate at higher OLRs. Interestingly, at the highest pre-concentration factor (2.30 g Na+ L−1) the difference between the permeate and the digester soluble COD indicated that membrane biofilm also played a role in COD removal. Finally, a preliminary energy and economic analysis showed that, at a pre-concentration factor of 10, the AnMBR temperature could be increased 10 °C and achieve a positive net present value (NPV) of 4 M€ for a newly constructed AnMBR treating 10,000 m3 d−1 of pre-concentrated sewage with an AnMBR lifetime of 20 years.

Development of clone with novel TrpE fusion tag in E. coli for overexpression of trypsin in a bench-scale bioreactor

Trypsin is a key enzyme under the serine proteases that is found in the pancreas which plays a key role in protein digestion. It cleaves peptide chains mainly at the carboxyl side of the amino acids lysine or arginine. This enzyme has received greater attention mainly due to its increased use in the removal of fusion tags during protein purification and its role in the processing of biosimilars like insulin. The present study was carried out to develop a clone with Novel TrpLE1413(TrpE) Fusion Tag for enhanced expression of trypsin which helps in cost reduction of biosimilar processing. In our experiment we have used a synthetic bovine trypsin gene containing a novel fusion tag TrpE at its N terminus, which was cloned into the pET41b (+) vector and expressed in E. coli BL21 (DE3) in a lab-scale bioreactor. Using the optimized fermentation process with TrpE Fusion Tag, 27.8 g/L inclusion bodies were produced at the end of fermentation, of which 209 mg/L of active trypsin was obtained after purification. In contrast, previous reports have claimed to produce a maximum of 60 mg/L of the enzyme without the fusion tag. Thus based on our findings, the small size (less than 2 kDa) of TrpE tag and its hydrophobicity may reduce the loss incurred during the purification process. Hence, it could be discerned that the use of the TrpE fusion tag along with a robust fermentation process led to 3– 4 fold higher yield making it a commercially viable process facilitating an improved recovery of the enzyme.

Effective nitrogen removal of wastewater from vitamin B2 production by a potential anammox process

The wastewater of vitamin B2 (VB2) production treated after anaerobic digesters contains high total nitrogen (TN) and low organic carbon. To efficiently remove nitrogen pollution in the wastewater, one lab-scale anaerobic ammonium oxidation (anammox) bioreactor was designed. Synthetic wastewater and proper conditions were used to startup the anammox process in the bioreactor. The stoichiometric parameters of the bioreactor showed that anammox process was successfully established during operation. Evaluation of the bioreactor with real VB2 wastewater suggested that the bioreactor can efficiently remove nitrogen pollution at low organic carbon situation. The optimal state of total nitrogen loading rate (TNLR) and the highest total nitrogen removal efficiency (TNRE) of the bioreactor were 0.35 kg/(m3·d) and 75%, respectively. The microbiota run at day 112 (named VBM112) was significantly different with the microbiotas of the two inoculation cultures. At the phylum level, the most abundant phylum was Proteobacteria in the inoculation culture, while Bacteroidetes was the most abundant phylum in VBM112. At the genus level, the composition of nitrogen removal related microbes significantly increased in VBM112. Especially, the anammox bacteria Candidatus Kuenenia increased from 0% of the inoculation culture to 2.2% of VBM112. The analyses of running parameters and microbiota profiles confirmed the well establishment of one lab-scale anammox bioreactor, which might be helpful for future full-scale VB2 wastewater treatment startup.

Improving bioremediation process of petroleum wastewater using biosurfactants producing Stenotrophomonas sp. S1VKR-26 and assessment of phytotoxicity

Primarily, this study aims to evaluate the biosurfactant production capability of Stenotrophomonas sp. S1VKR-26, profiling of its bioremediation ability to remediate petroleum refinery wastewater in a lab-scale bioreactor and assessment of phytotoxicity of bioremediated petroleum wastewater. As a result, strain S1VKR-26 was found to produce 5.15 g L−1 biosurfactant, CMC of 30 mg L−1 and reduced the surface tension from 60.3 to 30.5 mN m−1. Different PAHs like naphthalene (93%), phenanthrene (86%), fluoranthene (92%), and pyrene (98.3%), total petroleum hydrocarbons (72.33%) and phenolic compounds (93.06%) were significantly remediated from the wastewater after the treatment of strain S1VKR-26. Moreover, S1VKR-26 strain treated 1:1 diluted petroleum wastewater have higher germination (100%), vigor (486), and seedling (4.86 cm) compared to untreated wastewater. Therefore, the treatment of petroleum refinery wastewater with strain S1VKR-26 could be more effective in the sense of environmental safety and irrigation for crop production in agriculture.

Oxygen transfer rate affect polyhydroxybutyrate production and oxidative stress response in submerged cultures of Rhizobium phaseoli

Oxygen is a key factor during free-state growth of Rhizobium phaseoli or symbiosis with common bean roots. Five different initial oxygen transfer rates (OTRI) were fix by dissolved oxygen control and constants stirrer speed rates to determine the growth of R. phaseoli in a lab-scale bioreactor. The biomass did not presented differences in the maximum concentrations for OTRI of 1.0, 1.7 and 5.9 mmol l−1 h−1. However, a sublethal effect was observed for 11.9 and 20.2 mmol l−1 h−1, where lower biomass, kinetic and stoichiometric parameters were determined. Catalase activity as an oxidative stress indicator was approximately two-fold greater with 20.2 mmol l−1 h−1 than lower OTRI. Polyhydroxybutyrate (PHB) production did not present differences in the maximum concentration. Nevertheless, an increase in PHB accumulation, yield on biomass and specific productivity were determined at the higher OTRI, suggesting a defense mechanism against oxidative stress induce by the enhancement of the oxygen transfer rates.

TISSUE ENGINEERING BIOREACTORS: POTENTIAL APPLICATIONS AND SCALE UP STRATEGY

Tissue engineering bioreactors have been used in order to achieve production of artificial tissue, increasing cell proliferation capacity and yield and/or in vitro tissue/disease modelling. Although it is still discussing how to obtain functional and vascular tissue with these bioreactors, preclinical and clinical studies are ongoing. Tissue engineering bioreactors have been used as lab-scale bioreactors until now. Crucial potential application areas can be created by increasing the production capacity and bioprocess efficiency of these bioreactors. In this review, recent bioreactor technologies such as spinner flask, rotating wall/bed, hollow fiber membrane, perfusion and mechanical stimuli bioreactors are briefly presented in terms of their potential applications in medical field especially in the scope of scale-up approaches such as bubble column, stirred tank, membrane, air lift, fluidized packed and bed bioreactors.&#x0D; Peer Review History: &#x0D; Received 1 May 2020; Revised 11 June; Accepted 6 July, Available online 15 July 2020&#x0D; UJPR follows the most transparent and toughest ‘Advanced OPEN peer review’ system. The identity of the authors and, reviewers will be known to each other. This transparent process will help to eradicate any possible malicious/purposeful interference by any person (publishing staff, reviewer, editor, author, etc) during peer review. As a result of this unique system, all reviewers will get their due recognition and respect, once their names are published in the papers. We expect that, by publishing peer review reports with published papers, will be helpful to many authors for drafting their article according to the specifications. Auhors will remove any error of their article and they will improve their article(s) according to the previous reports displayed with published article(s). The main purpose of it is ‘to improve the quality of a candidate manuscript’. Our reviewers check the ‘strength and weakness of a manuscript honestly’. There will increase in the perfection, and transparency.&#x0D; Received file &#x0D; &#x0D; Average Peer review marks at initial stage: 6.0/10&#x0D; Average Peer review marks at publication stage: 8.0/10&#x0D; Reviewer(s) detail:&#x0D; Name: Dr. Sally A. El-Zahaby&#x0D; Affiliation: Pharos University in Alexandria, Egypt&#x0D; E-mail: sally.elzahaby@yahoo.com&#x0D; &#x0D; Name: DR Viney Chawla&#x0D; Affiliation: University Institute of Pharmaceutical Sciences&#x0D; Baba Farid University of Health Sciences&#x0D; Sadiq Road, Faridkot-Punjab 151203&#x0D; E-mail: drvineychawla@gmail.com&#x0D; &#x0D; Comments of reviewer(s): &#x0D; &#x0D;

Response surface methodology based optimization and scale-up production of amylase from a novel bacterial strain, Bacillus aryabhattai KIIT BE-1

A novel strain KIIT BE-1 isolated from a specialized environment, screened through starch iodine test from a set of eighty-five biodigestate isolates, produced amylase maximally when cultured for 48 h at 37 °C. The molecular and biochemical characterization confirmed it as a strain of Bacillus aryabhattai. It exhibited optimal amylase activity (3.20 U/ml) at 36 h post incubation with a media combination of starch and yeast extract for C-N source respectively. Statistical optimisation by response surface modeling showed R2 values of 0.9645 for biomass and 0.9831 for amylase activity, suggesting the significance of the model. The optimised medium (10.25% starch, 5.0% peptone, 5.18% yeast extract, pH 7.3) enhanced the enzyme activity to 4.16 U/ml (1.39-fold) from 3.20 U/ml of un-optimised medium. Further, the biomass yield and the enzymatic activity in optimized medium and process conditions increased by 1.14 and 1.21 folds subjected to a 5 l scaled-up operation in a lab-scale bioreactor.

Zero-valent iron addition in anaerobic dynamic membrane bioreactors for preconcentrated wastewater treatment: Performance and impact

Wastewater preconcentration to capture abundant organics is promising for facilitating subsequent anaerobic digestion (AD) to recover bioenergy, however research efforts are still needed to verify the effectiveness of such an emerging strategy as carbon capture plus AD. Therefore, lab-scale anaerobic dynamic membrane bioreactors (AnDMBRs) without and with the addition of zero-valent iron (ZVI) (i.e., AnDMBR1 versus AnDMBR2) were developed for preconcentrated domestic wastewater (PDW) treatment, and the impact of ZVI addition on process performance and associated mechanisms were investigated. The stepwise addition of ZVI from 2 to 4 to 6 g/L improved the treatment performance as COD removal slightly increased and TP removal and methane production were enhanced by 53.3%–62.9% and 22.6%–31.3%, respectively, in consecutive operational phases. However, the average increasing rate of the transmembrane pressure (TMP) in AnDMBR2 (0.18 kPa/d) was obviously higher than that in AnDMBR1 (0.05 kPa/d), indicating an unfavorable impact of dosing ZVI on the dynamic membrane (DM) filtration performance. ZVI that has transformed to iron ions (mainly Fe2+) can behave as a coagulant, electron donor or inorganic foulant, thus enabling the excellent removal of dissolved phosphorous, enhancing the enrichment and activities of specific methanogens and causing the formation of a compact DM layer. Morphological, componential, and microbial community analyses provided new insights into the functional mechanisms of ZVI added to membrane-assisted anaerobic digesters, indicating that ZVI has the potential to improve bioenergy production and resource recovery, while optimizing the ZVI dosage should be considered to alleviate membrane fouling.

Generic Model Control Applied to E. coli BL21(DE3) Fed-Batch Cultures

This work proposes a Generic Model Control (GMC) strategy to regulate biomass growth in fed-batch cultures of Escherichia coli BL21(DE3). The control law is established using a previously validated mechanistic model based on the overflow metabolism paradigm. A model reduction is carried out to prevent the controller from relying on kinetics, which may be uncertain. In order to limit the controller to the use of a single measurement, i.e., biomass concentration which is readily available, a Kalman filter is designed to reconstruct the nonmeasurable information from the outlet gas and the remaining stoichiometry. Several numerical simulations are presented to assess the controller robustness with respect to model uncertainty. Experimental validation of the proposed GMC strategy is achieved with a lab-scale bioreactor.

A biorefinery for conversion of citrus peel waste into essential oils, pectin, fertilizer and succinic acid via different fermentation strategies

A process for the valorization of citrus peel waste (CPW) has been developed aiming to produce succinic acid and a series of added-value products through the biorefinery platform. CPW was subject to physicochemical and biological treatment to isolate essential oils (0.43%) and pectin (30.53%) as extractable products, pretreating the material for subsequent production of succinic acid that enabled application of remaining biorefinery residues (BR) as fertilizer substitute. Cellulose, hemicellulose and lignin contents of CPW accounted for 22.45%, 8.05% and 0.66% respectively, while acid hydrolysis reduced hemicellulose by 3.42% in BR. Moreover, essential oils extracted from CPW included 17 compounds, among which D-limonene reached 96.7%. The hydrolyzate generated was fermented for succinic acid production using Actinobacillus succinogenes. Different batch experiments demonstrated that the combined use of corn steep liquor (CSL) and vitamins in a lab-scale bioreactor resulted in product concentration and yield that reached 18.5 g L−1 and 0.62 g g−1 respectively. Although simultaneous saccharification and fermentation (SSF) could not enhance succinic acid production, a fed-batch fermentation strategy increased succinic acid concentration and yield generating 22.4 g L−1 and 0.73 g g−1 respectively, while the mass of the platform chemical formed was enhanced by 27% as compared to the batch process. BR was explored as fertilizer substitute aiming to close the loop in the management of CPW towards development of a zero-waste process demonstrating that although the material imposed stress on plant growth, the content of potassium, phosphorus and nitrogen in the mixture increased.

Respirometric techniques coupled with laboratory-scale tests for kinetic and stoichiometric characterisation of fungal and bacterial tannin-degrading biofilms.

In environmental biotechnology applications for wastewater treatment, bacterial-based bioprocesses are mostly implemented; on the contrary, the application of fungal-based bioprocesses, is still challenging under non-sterile conditions. In a previous laboratory-scale study, we showed that when specific tannins are used as the sole carbon source, fungi can play a key role in the microbial community, under non-sterile conditions and in the long term. In a previous study, an engineered ecosystem, based on fungal tannin biodegradation, was successfully tested in a laboratory-scale bioreactor under non-sterile conditions. In the present study, a kinetic and stoichiometric characterisation of the biomass developed therein was performed through the application of respirometric techniques applied to the biomass collected from the above-mentioned reactor. To this aim, a respirometric set-up was specifically adapted to obtain valuable information from tannin-degrading fungal biofilms. A mathematical model was also developed and applied to describe both the respirometric profiles and the experimental data collected from the laboratory-scale tests performed in the bioreactor. The microbial growth was described through a Monod-type kinetic equation as a first approach. Substrate inhibition, decay rate and tannin hydrolysis process were included to better describe the behaviour of immobilised biomass selected in the tannin-degrading bioreactor. The model was implemented in AQUASIM using the specific tool Biofilm Compartment to simulate the attached fungal biofilm. Biofilm features and transport parameters were either measured or assumed from the literature. Key kinetic and stoichiometric unknown parameters were successfully estimated, overcoming critical steps for scaling-up a novel fungal-based technology for tannins biodegradation.