Reactive extraction of lactic and acetic acids from leached bed reactor leachate and process optimization by response surface methodology.

The present work focused on extracting lactic and acetic acids from the leachate collected from leached bed reactor (LBR) during acidogenesis of food waste using the reactive extraction (RE) process. A wide range of diluents was screened either alone by physical extraction (PE) or in combination with extractants using RE to extract acids from the VFA mix. Aliquat 336-Butyl acetate/MIBK extractants in RE demonstrated higher distribution coefficients (k) and extraction yield (E %) than PE. The response surface methodology (RSM) was used to optimize the extraction of lactic and acetic acids from the synthetic acid mix, using three variables (extractant concentrations, solute/acid concentration and time). Consequently, these three variables were optimized for LBR leachate. The RE was promising, and extraction efficiencies of 65% (lactate), 75% (acetate), 86.2% (propionate) and almost 100% for butyrate and medium-chain fatty acids (MCFA) were achieved after 16 h of extraction. The RSM optimization predicted a maximum E % of 59.60% and 34.67% for lactate and acetate in 5.5 and 1.17 min, respectively. In the leachate experiment, an increase in E% and k was observed with increasing extractant concentration and lactate and acetate concentrations over time. Using a 1M reactive extractant mix and 1.25 and 12 g/L of solute concentrations, the maximum E % of acetate and lactate were 38.66% and 61.8% in 10 min. The results could contribute to developing a rapid in-situ product recovery system integrated with food waste acidogenesis for lactate and acetate recovery, contributing to the bio-economy.

Sustainable enzymatic treatment of organic waste in a framework of circular economy.

Enzymatic treatment of food and vegetable waste (FVW) is an eco-friendly approach for producing industrially relevant value-added products. This review describes the sources, activities and potential applications of crucial enzymes in FVW valorization. The specific roles of amylase, cellulase, xylanase, ligninase, protease, pectinase, tannase, lipase and zymase enzymes were explained. The exhaustive list of value-added products that could be produced from FVW is presented. FVW valorization through enzymatic and whole-cell enzymatic valorization was compared. The note on global firms specialized in enzyme production reiterates the economic importance of enzymatic treatment. This review provides information on choosing an efficient enzymatic FVW treatment strategy, such as nanoenzyme and cross-linked based enzyme immobilization, to make the process viable, sustainable and cheaper. Finally, the importance of life cycle assessment of enzymatic valorization of FVW was impressed to prove this approach is a better option to shift from a linear to a circular economy.

Nonlinear electrostatic ion cyclotron wave collapse and formation of wave packets in the presence of trapped electrons.

The weakly nonlinear and dispersive electrostatic ion cyclotron wave dynamics in the presence of Schamel distributed trapped electrons is studied in collisionless plasmas. The dynamics of the nonlinear wave is shown to be governed by a Schamel-Ostrovsky type equation. Analytical and numerical solutions of this equation reveal the collapse of a solitary (localized) pulse at a critical time that depends on the trapping parameter and the strength of the magnetic field. The time-dependent computational result is noteworthy, which predicts the formation of wave packets (wave group) beyond the critical time. The results are in good agreement with the astrophysical observations in auroral plasmas.

Two-phase anaerobic digestion of food waste: Effect of semi-continuous feeding on acidogenesis and methane production.

In present investigation, effect of diverting acidogenic off-gas from leached bed reactor (LBR) to up-flow anaerobic sludge blanket (UASB) reactor during semi-continuous food waste (FW) anaerobic digestion was evaluated. In test LBR headspace pressure (3.3 psi) was maintained with intermittent headspace gas transfer into UASB. In control, same headspace pressure was maintained without gas transfer. The semi-continuous FW addition affected the characteristics and production of leachate in control and test LBR. The cumulative COD, total soluble products and methane yields were 1.26, 1.37 and 3 times higher in the test LBR than the control. The acetate and methane yields from test LBR were 697.8 g·kgVSadded-1 and 167.55 mL·gCOD-1feeding. Acidogenic gas transfer maintained low partial pressure of hydrogen and the hydrogen to carbon-di-oxide ratio in the headspace of LBR, which were thermodynamically favorable for microbial metabolism and concomitant high-rate production of acetate-rich volatile fatty acid and methane-rich biogas from FW.

Food waste and sewage sludge co-digestion amended with different biochars: VFA kinetics, methane yield and digestate quality assessment.

This work investigated the impact of the addition of different biochar types on mitigation of volatile fatty acid (VFA) accumulation, methane recovery and digestate quality in mesophilic food waste-sludge co-digestion. Four biochars derived from agricultural and sludge residues under different pyrolysis temperatures were compared. Specific biochar properties such as pH, surface area, chemical properties and presence of surface functional groups likely influenced biochar reactions during digestion, thereby resulting in a varying performance of different biochars. Miscanthus straw biochar addition led to the highest specific methane yield of 307 ± 0.3 mL CH4/g VSadded versus 241.87 ± 5.9 mL CH4/g VSadded from control with no biochar addition over 30 days of the co-digestion period. Biochar supplementation led to enhanced process stability which likely resulted from improved syntrophic VFA oxidation facilitated by specific biochar properties. Overall, a 21.4% increase in the overall methane production was obtained with biochar addition as compared to control. The resulting digestate quality was also investigated. Biochar-amended digester generated a digestate rich in macro- and micro-nutrients including K, Mg, Ca, Fe making biochar-amended digestate a potential replacement of agricultural lime fertilizer. This work demonstrated that the addition of specific biochars with desirable properties alleviated VFA accumulation and facilitated enhanced methane recovery, thereby providing a means to achieve process stability even under high organic loading conditions in co-digestions. Moreover, the availability of biochar-enriched digestate with superior characteristics than biochar-free digestate adds further merit to this process.

Algal biorefinery models with self-sustainable closed loop approach: Trends and prospective for blue-bioeconomy.

Microalgae due to its metabolic versatility have received a focal attention in the biorefinery and bioeconomy context. Microalgae products have broad and promising application potential in the domain of renewable fuels/energy, nutraceutical, pharmaceuticals and cosmetics. Biorefining of microalgal biomass in a circular loop with an aim to maximize resource recovery is being considered as one of the sustainable option that will have both economical and environmental viability. The expansive scope of microalgae cultivation with self-sustainability approach was discussed in this communication in the framework of blue-bioeconomy. Microalgae based primary products, cultivation strategies, valorization of microalgae biomass for secondary products and integrated biorefinery models for the production of multi-based products were discussed. The need and prospect of self-sustainable models in closed loop format was also elaborated.

Efficient resource valorization by co-digestion of food and vegetable waste using three stage integrated bioprocess.

During two-stage (Acidogenesis-Methanogenesis) process, solid organics and gaseous by-products are usually left unused. To increase resource recovery efficiency, a three stage process (Hydrolysis/Acidogenesis-Methanogenesis-Composting) was designed. Initially, co-digestion of food waste (FW) and vegetable waste (VW) was carried out in Leach Bed Reactor (LBR) for hydrolysis and acidogenesis, followed by airlift reactor (ALR) for methanogenesis for 21 days using two different feed stocks [2:3 FW:VW~FVW; FW alone]. Off gas from LBR was diverted to ALR to enhance methane recovery. Results depicted that volatile fatty acids (VFA) and biohydrogen production was more for FW fed system, while methane production was higher in FVW fed system. Three different functional zones in three separate chambers significantly accelerated organic removal rate while gas diversion increased overall methane recovery. In third stage, residual solid organic matter from LBR was subjected to aerobic composting and compost with N (%): 2.90 & 2.76; C/N ratio: 18.2 & 20.8 for FVW and FW was recovered. The three-stage process has advantages of zero waste generation and overall process stability, accounting for resource efficient circular loop.

Integration of biological pre-treatment methods for increased energy recovery from paper and pulp biosludge.

The paper and pulp industry (PPI) produces high quantities of solid and liquid discharge and is regarded as the most polluting industry in the world causing adverse effects to environments and human beings. Hence changes in the way PPI sludge and waste materials are treated is urgently required. Nearly, 10 million tons of waste is generated per year, however PPI waste is enriched with many organic chemicalscontaining a high percentage of lignin, cellulose, and hemicellulose which can be used as valuable raw materials for the production of bioenergy and value-added chemicals. Pretreatment of complex lignocellulosic materials of PPI waste is difficult because of the cellulose crystallinity and lignin barrier. At present most of this waste is recycled in a conventional treatment approach through biological and chemical processes, incurring high cost and low returns. Henceefficient pretreatment techniques are required by which complete conversion of PPI waste is possible. Therefore, the present chapter provides the scope of integration of pretreatment methods through which bioenergy recovery is possible during the PPI waste treatment. Detailed information is presented on the various pre-treatment techniques (chemical, mechanical, enzymatic and biological) in order to increase the efficiency of PPI waste treatment and energy recovery from PPI waste. Along with acid and alkali based efficient chemical treatment process, physical methods (i.e. shearing, high-pressure homogenization, etc.), biochemical techniques (whole cell-based and enzyme-based) and finally biological techniques (e.g. aerobic and anaerobic treatment) are discussed. During each of the treatment processes, scope of energy recovery and bottlenecks of the processes were elaborated. The review thus provides systemic insight into developing efficient pretreatment processes which could increase carbon recovery and treatment efficiency of PPI waste.

Effect of food to vegetable waste ratio on acidogenesis and methanogenesis during two-stage integration.

The mixing ratio of food waste (FW) to vegetable waste (VW) (2:3 FW:VW ∼ 152.51 g VS and 2:1 FW:VW ∼ 137.03 gVS) was optimized using two-stage (LBR-UASB) experimental process depending upon volatile solid (VS) load. The effect of FW to VW ratio was studied in Leach Bed Reactor (LBR) towards leachate production. Results revealed that hydrolysis rate (73.11%), sCOD (3294.3 g/KgVS) and tVFA (2664 g/KgVS) yield was higher in 2:1 FW:VW ratio. Acetate, propionate, lactate and methane yield for 2:3 FW:VW (420 g/KgVS, 87 g/KgVS, 180 g/KgVS and 226.86 ml/gVS respectively) was different from 2:1 FW:VW (340 g/KgVS, 247 g/KgVS, 340 g/KgVS and 218.54 ml/gVS respectively). 2:3 FW:VW ratio depicted higher VS (53.96%) and COD (54.1%) removal than 2:1 FW:VW ratio 46.34% and 41.8% respectively. VW addition regulated pH, restricted propionate and lactate production with enhanced methanogenesis by improving acetate production in two-stage AD process which further boosted process stability and efficiency.

Application of recombinant Pediococcus acidilactici BD16 (fcs +/ech +) for bioconversion of agrowaste to vanillin.

Biotechnological production of vanillin is gaining momentum as the natural synthesis of vanillin that is very expensive. Ferulic acid (FA), a costly compound, is used as the substrate to produce vanillin biotechnologically and the making process is still expensive. Therefore, this study investigated the practical use of an agrobiomass waste, rice bran, and provides the first evidence of a cost-effective production of vanillin within 24 h of incubation using recombinant Pediococcus acidilactici BD16 (fcs +/ech +). Introduction of two genes encoding feruloyl CoA synthetase and enoyl CoA hydratase into the native strain increased vanillin yield to 4.01 g L-1. Bioconversion was monitored through the transformation of phenolic compounds. A hypothetical metabolic pathway of rice bran during the vanillin bioconversion was proposed with the inserted pathway from ferulic acid to vanillin and compared with that of other metabolic engineered strains. These results could be a gateway of using recombinant lactic acid bacteria for industrial production of vanillin from agricultural waste.

Agrowaste to vanillin conversion by a natural Pediococcus acidilactici strain BD16.

Owing to its flavoring, antimicrobial, antioxidant and anticarcinogenic nature, vanillin is widely used in foods, beverages, perfumes and pharmaceutical products. Ferulic acid (FA) is an important precursor of vanillin which is abundant in cereals like maize, rice and wheat and sugar beet. A major drawback of microbial vanillin production from FA is the degradation and biotransformation of toxic vanillin to other phenolic derivatives. The present study is undertaken to explore microbial vanillin production from FA precursor rice bran by employing vanillin-resistant Pediococcus acidilactici BD16, a natural lactic acid bacteria isolate. Extracellular, intracellular and cellular vanillin dehydrogenase activity was found least, which was minimized vanillin degradation, and the strain resists more than 5 g L-1 vanillin in the medium. A metabolomics approach was followed for the detection of FA, vanillin and other metabolites generated during fermentation of rice bran medium. A metabolic pathway was also predicted for vanillin biosynthesis. Approximately 1.06 g L-1 of crude vanillin was recovered from rice-bran-containing medium and this further offers scope for the industrial utilization of the organism and its genetic manipulation to enhance production of biovanillin.

Metabolic engineering of E. coli top 10 for production of vanillin through FA catabolic pathway and bioprocess optimization using RSM.

Metabolic engineering and construction of recombinant Escherichia coli strains carrying feruloyl-CoA synthetase and enoyl-CoA hydratase genes for the bioconversion of ferulic acid to vanillin offers an alternative way to produce vanillin. Isolation and designing of fcs and ech genes was carried out using computer assisted protocol and the designed vanillin biosynthetic gene cassette was cloned in pCCIBAC expression vector for introduction in E. coli top 10. Recombinant strain was implemented for the statistical optimization of process parameters influencing F A to vanillin biotransformation. CCD matrix constituted of process variables like FA concentration, time, temperature and biomass with intracellular, extracellular and total vanillin productions as responses. Production was scaled up and 68 mg/L of vanillin was recovered from 10 mg/L of FA using cell extracts from 1 mg biomass within 30 min. Kinetic activity of enzymes were characterized. From LCMS-ESI analysis a metabolic pathway of FA degradation and vanillin production was predicted.

Anaerobic co-digestion of food waste and chemically enhanced primary-treated sludge under mesophilic and thermophilic conditions.

Anaerobic co-digestion of food waste with primary sewage sludge is beneficial for urban centers, while the optimized conditions reported in the literature are not locally suitable for Hong Kong. Therefore, the present study was aimed to develop an optimized mixing ratio of food waste to chemically enhanced primary-treated sewer sludge (CEPT) for co-digestion using batch tests under mesophilic (37°C) and thermophilic (55°C) conditions. The mixing ratios of 1:1, 1:2, 1:3, 2:1 and 3:1 (v v(-1)) of food waste to CEPT sludge was tested under the following conditions: temperature - 35°C and 55°C; pH - not regulated; agitation - 150 rpm and time - 20 days. The thermophilic incubations led a good hydrolysis rate and 2-12-fold higher enzyme activities than in mesophilic incubations for different mixing ratios. While the acidogenesis were found retarded that leading to 'sour and stuck' digestion for all mixing ratio of food waste to CEPT sludge from thermophilic incubations. The measured zeta potential was most favourable (-5 to -16.8 mV) for methane production under thermophilic incubations; however the CH4 recovery was less than that in mesophilic incubations. The results suggested that the quick hydrolysis and subsequent acid accumulation under thermophilic incubation lead to inhibited methanogenesis at the early stage than in mesophilic systems. It is concluded that buffer addition is therefore required for any mixing ratio of food waste to CEPT sludge for improved CH4 recovery for both mesophilic and thermophilic operations.

Application of recombinant Pediococcus acidilactici BD16 (fcs⁺/ech⁺) in malolactic fermentation.

This study was conducted to enhance flavor characteristics of wine by malolactic fermentation using recombinant Pediococcus acidilactici BD16 (fcs (+)/ech (+)) encoding synthetic genes of feruloyl-CoA synthetase and enoyl-CoA hydratase. After malolactic fermentation, wine phenolics were characterized using LCMS-ESI technique and a significant improvement in the antioxidant activity and flavor characteristics of wine was observed due to increased concentration of cinnamic acid derivatives. This proof of concept study highlights the role of recombinant P. acidilactici BD16 (fcs (+)/ech (+)) in improving flavor as well as aroma of wine due to production of several phenolic derivatives during secondary fermentation. A novel metabolic pathway was predicted from mass spectral analysis data that indicates biotransformation of cinnamic acid and derivatives into apigenin, catechin, coniferyl aldehyde, cyanidin, hydroxybenzoic acids, laricitrin, luteolin, malvidin 3-glucoside, myricetin, naringenin, pelargonin, piceatannol, querecitin, and vanillin that not only increased the overall consumer appreciation but also improved nutritional and probably the therapeutic properties of wines. This is a first evidence-based study where role of recombinant P. acidilactici BD16 (fcs (+)/ech (+)) in the wine secondary fermentation has been elucidated.

Metabolic engineering of Pediococcus acidilactici BD16 for production of vanillin through ferulic acid catabolic pathway and process optimization using response surface methodology.

Occurrence of feruloyl-CoA synthetase (fcs) and enoyl-CoA hydratase (ech) genes responsible for the bioconversion of ferulic acid to vanillin have been reported and characterized from Amycolatopsis sp., Streptomyces sp., and Pseudomonas sp. Attempts have been made to express these genes in Escherichia coli DH5α, E. coli JM109, and Pseudomonas fluorescens. However, none of the lactic acid bacteria strain having GRAS status was previously proposed for heterologous expression of fcs and ech genes for production of vanillin through biotechnological process. Present study reports heterologous expression of vanillin synthetic gene cassette bearing fcs and ech genes in a dairy isolate Pediococcus acidilactici BD16. After metabolic engineering, statistical optimization of process parameters that influence ferulic acid to vanillin biotransformation in the recombinant strain was carried out using central composite design of response surface methodology. After scale-up of the process, 3.14 mM vanillin was recovered from 1.08 mM ferulic acid per milligram of recombinant cell biomass within 20 min of biotransformation. From LCMS-ESI spectral analysis, a metabolic pathway of phenolic biotransformations was predicted in the recombinant P. acidilactici BD16 (fcs (+)/ech (+)).

Phenolic biotransformations during conversion of ferulic acid to vanillin by lactic acid bacteria.

Vanillin is widely used as food additive and as a masking agent in various pharmaceutical formulations. Ferulic acid is an important precursor of vanillin that is available in abundance in cell walls of cereals like wheat, corn, and rice. Phenolic biotransformations can occur during growth of lactic acid bacteria (LAB), and their production can be made feasible using specialized LAB strains that have been reported to produce ferulic acid esterases. The present study aimed at screening a panel of LAB isolates for their ability to release phenolics from agrowaste materials like rice bran and their biotransformation to industrially important compounds such as ferulic acid, 4-ethyl phenol, vanillic acid, vanillin, and vanillyl alcohol. Bacterial isolates were evaluated using ferulic acid esterase, ferulic acid decarboxylase, and vanillin dehydrogenase assays. This work highlights the importance of lactic acid bacteria in phenolic biotransformations for the development of food grade flavours and additives.

Biotransformation of rice bran to ferulic acid by pediococcal isolates.

Ferulic acid (FA) is widely used in foods, in beverages, and in various pharmaceutical industries as a precursor of vanillin. FA biotransformation can occur during the growth of lactic acid bacteria (LAB), and its conversion to other phenolic derivatives is observed by many scientists, where ferulic acid esterase (FAE) and ferulic acid decarboxylase (FDC) play significant roles. The present study aimed at screening a panel of LAB for their ability to release FA from rice bran, an agro waste material. FAE and FDC activities were analyzed for the preliminary screening of various dairy isolates. Two Pediococcus acidilactici isolates were selected for studying further the hydrolysis of FA from rice bran and its bioconversion into phenolic derivatives like 4-ethylphenol, vanillin, vanillic acid, and vanillyl alcohol. P. acidilactici M16, a probiotic isolate, has great potential for the production of FA from rice bran and could be exploited as starter culture in the food industry for the production of biovanillin.

Biotechnological and molecular approaches for vanillin production: a review.

Vanillin is one of the most widely used flavoring agents in the world. As the annual world market demand of vanillin could not be met by natural extraction, chemical synthesis, or tissue culture technology, thus biotechnological approaches may be replacement routes to make production of bio-vanillin economically viable. This review's main focus is to highlight significant aspects of biotechnology with emphasis on the production of vanillin from eugenol, isoeugenol, lignin, ferulic acid, sugars, phenolic stilbenes, vanillic acid, aromatic amino acids, and waste residues by applying fungi, bacteria, and plant cells. Production of biovanillin using GRAS lactic acid bacteria and metabolically engineered microorganisms, genetic organization of vanillin biosynthesis operons/gene cassettes and finally the stability of biovanillin generated through various biotechnological procedures are also critically reviewed in the later sections of the review.

Statistical media and process optimization for biotransformation of rice bran to vanillin using Pediococcus acidilactici.

An isolate of P. acidilactici capable of producing vanillin from rice bran was isolated from a milk product. Response Surface Methodology was employed for statistical media and process optimization for production of biovanillin. Statistical medium optimization was done in two steps involving Placket Burman Design and Central Composite Response Designs. The RSM optimized vanillin production medium consisted of 15% (w/v) rice bran, 0.5% (w/v) peptone, 0.1% (w/v) ammonium nitrate, 0.005% (w/v) ferulic acid, 0.005% (w/v) magnesium sulphate, and 0.1% (v/v) tween-80, pH 5.6, at a temperature of 37 degrees C under shaking conditions at 180 rpm. 1.269 g/L vanillin was obtained within 24 h of incubation in optimized culture medium. This is the first report indicating such a high vanillin yield obtained during biotransformation of ferulic acid to vanillin using a Pediococcal isolate.