Challenges and advancements in bioprocess intensification of fungal secondary metabolite: kojic acid.

Kojic acid is a fungal secondary metabolite commonly known as a tyrosinase inhibitor, that acts as a skin-whitening agent. Its applications are widely distributed in the area of cosmetics, medicine, food, and chemical synthesis. Renewable resources are the alternative feedstocks that can fulfill the demand for free sugars which are fermented for the production of kojic acid. This review highlights the current progress and importance of bioprocessing of kojic acid from various types of competitive and non-competitive renewable feedstocks. The bioprocessing advancements, secondary metabolic pathway networks, gene clusters and regulations, strain improvement, and process design have also been discussed. The importance of nitrogen sources, amino acids, ions, agitation, and pH has been summarized. Two fungal species Aspergillus flavus and Aspergillus oryzae are found to be extensively studied for kojic acid production due to their versatile substrate utilization and high titer ability. The potential of A. flavus to be a competitive industrial strain for large-scale production of kojic acid has been studied.

The impact of carbon and nitrogen catabolite repression in microorganisms.

Organisms have cellular machinery that is focused on optimum utilization of resources to maximize growth and survival depending on various environmental and developmental factors. Catabolite repression is a strategy utilized by various species of bacteria and fungi to accommodate changes in the environment such as the depletion of resources, or an abundance of less-favored nutrient sources. Catabolite repression allows for the rapid use of certain substrates like glucose over other carbon sources. Effective handling of carbon and nitrogen catabolite repression in microorganisms is crucial to outcompete others in nutrient limiting conditions. Investigations into genes and proteins linked to preferential uptake of different nutrients under various environmental conditions can aid in identifying regulatory mechanisms that are crucial for optimum growth and survival of microorganisms. The exact time and way bacteria and fungi switch their utilization of certain nutrients is of great interest for scientific, industrial, and clinical reasons. Catabolite repression is of great significance for industrial applications that rely on microorganisms for the generation of valuable bio-products. The impact catabolite repression has on virulence of pathogenic bacteria and fungi and disease progression in hosts makes it important area of interest in medical research for the prevention of diseases and developing new treatment strategies. Regulatory networks under catabolite repression exemplify the flexibility and the tremendous diversity that is found in microorganisms and provides an impetus for newer insights into these networks.

Carbon, nitrogen and phosphorus removal mechanisms of aerobic granules.

Aerobic granules are the potential tools to develop modern wastewater treatment technologies with improved nutrient removal efficiency. These granules have several promising advantages over conventional activated sludge-based wastewater treatment processes. This technology has the potential of reducing the infrastructure and operation costs of wastewater treatment by 25%, energy requirement by 30%, and space requirement by 75%. The nutrient removal mechanisms of aerobic granules are slightly different from that of the activated sludge. For instance, unlike activated sludge process, according to some reports, as high as 70% of the total phosphorus removed by aerobic granules were attributed to precipitation within the granules. Similarly, aerobic granule-based technology reduces the total amount of sludge produced during wastewater treatment. However, the reason behind this observation is unknown and it needs further explanations based on carbon and nitrogen removal mechanisms. Thus, as a part of the present review, a set of new hypotheses have been proposed to explain the peculiar nutrient removal mechanisms of the aerobic granules.

Hydrolytic pre-treatment methods for enhanced biobutanol production from agro-industrial wastes.

Brewery industry liquid waste (BLW), brewery spent grain (BSG), apple pomace solid wastes (APS), apple pomace ultrafiltration sludge (APUS) and starch industry wastewater (SIW) have been considered as substrates to produce biobutanol. Efficiency of hydrolysis techniques tested to produce fermentable sugars depended on nature of agro-industrial wastes and process conditions. Acid-catalysed hydrolysis of BLW and BSG gave a total reducing sugar yield of 0.433 g/g and 0.468 g/g respectively. Reducing sugar yield from microwave assisted hydrothermal method was 0.404 g/g from APS and 0.631 g/g from APUS, and, 0.359 g/g from microwave assisted acid-catalysed SIW dry mass. Parameter optimization (time, pH and substrate concentration) for acid-catalysed BLW hydrolysate utilization using central composite model technique produced 307.9 g/kg glucose with generation of inhibitors (5-hydroxymethyl furfural (20 g/kg), furfural (1.6 g/kg), levulinic acid (9.3 g/kg) and total phenolic compound (0.567 g/kg)). 10.62 g/L of acetone-butanol-ethanol was produced by subsequent clostridial fermentation of the substrate.

Acid mediated chemical treatment to remove sugar from waste acid stream from nano-crystalline cellulose manufacturing process.

Nano-crystalline cellulose (NCC) is a nano-scale biomaterial derived from highly abundant natural polymer cellulose. It is industrially produced by concentrated acid hydrolysis of cellulosic materials. However, presences of as high as 5-10% of sugar monomers in spent sulphuric acid during the manufacturing process, makes it unsuitable for such recycling or reuse of sulphuric acid. Currently, the industry has been using membrane and ion exchange technology to remove such sugars, however, such technologies cannot achieve the target of 80-90% removal. In the current investigation, thermal treatment and acid mediated thermal treatment have been evaluated for sugar removal from the spent sulphuric acid. Almost complete removal of sugar has been achieved by this approach. Maximum sugar removal efficiency (99.9%) observed during this study was at 120±1°C for 60min using 0.8 ratio (sample: acid) or at 100±1°C for 40min using 1.5 ratio.

Sustainable commercial nanocrystalline cellulose manufacturing process with acid recycling.

Nanocrystalline cellulose (NCC) is a biomaterial having potential applications in a wide range of industries. It is industrially produced by concentrated acid hydrolysis of cellulosic materials. In this process, the sulfuric acid rich liquor can be concentrated and reused. However, removal of sugar monomers and oligomers is necessary for such recycling. Membrane and ion exchange technology can be employed to remove sugars; however, such technologies are not efficient in meeting the quality required to recycle the acid solution. As a part of the present study, activated carbon (AC) has been evaluated as an adsorbent for sugar removal from the acidic solution generated during commercial nanocrystalline cellulose manufacturing process. Almost complete removal of sugar can be achieved by this approach. The maximum sugar removal observed during this study was 3.4g/g of AC. Based on this finding, a sustainable method has been proposed for commercial nanocrystalline cellulose manufacturing.

Finding Knowledge Gaps in Aerobic Granulation Technology.

This review identifies the knowledge gaps in aerobic granulation technology and defines some problems for future studies. In particular, extracellular polymeric substances (EPSs) should be further characterized to understand the intermolecular interactions among these polymers, the role of chelating agents in destabilizing EPS ionic bridges needs further elucidation, and early detection of the quorum-quenching enzymes should be considered to avoid granule segregation and process failure. Furthermore, the process should be supplemented with volatile fatty acids as electron donors/carbon sources, and appropriate anoxic/anaerobic conditions should be provided for enhanced nitrogen and phosphorus removal. Finally, the biodegradation, bioaccumulation, biosorption, and mass transfer behaviors of the emerging contaminants within the granules need further investigation.

Potential Application of Biohydrogen Production Liquid Waste as Phosphate Solubilizing Agent-A Study Using Soybean Plants.

With CO2 free emission and a gravimetric energy density higher than gasoline, diesel, biodiesel, and bioethanol, biohydrogen is a promising green renewable energy carrier. During fermentative hydrogen production, 60-70 % of the feedstock is converted to different by-products, dominated by organic acids. In the present investigation, a simple approach for value addition of hydrogen production liquid waste (HPLW) containing these compounds has been demonstrated. In soil, organic acids produced by phosphate solubilizing bacteria chelate the cations of insoluble inorganic phosphates (e.g., Ca3 (PO4)2) and make the phosphorus available to the plants. Organic acid-rich HPLW, therefore, has been evaluated as soil phosphate solubilizer. Application of HPLW as soil phosphate solubilizer was found to improve the phosphorus uptake of soybean plants by 2.18- to 2.74-folds. Additionally, 33-100 % increase in seed germination rate was also observed. Therefore, HPLW has the potential to be an alternative for phosphate solubilizing biofertilizers available in the market. Moreover, the strategy can be useful for phytoremediation of phosphorus-rich soil.

Biohydrogen production by co-fermentation of crude glycerol and apple pomace hydrolysate using co-culture of Enterobacter aerogenes and Clostridium butyricum.

Co-substrate utilization of various wastes with complementary characteristics can provide a complete medium for higher hydrogen production. This study evaluated potential of apple pomace hydrolysate (APH) co-fermented with crude glycerol (CG) for increased H2 production and decreased by-products formation. The central composite design (CCD) along with response surface methodology (RSM) was used as tool for optimization and 15 g/L of CG, 5 g/L of APH and 15% (v/v) inoculum were found to be optimum to produce as high as 26.07 ± 1.57 mmol H2/L of medium. The p-value of 0.0017 indicated that APH at lower concentration had a significant effect on H2 production. By using CG as sole carbon source, reductive pathway of glycerol metabolism was favored with 19.46 mmol H2/L. However, with APH, oxidative pathway was favored with higher H2 production (26.07 ± 1.57 mmol/L) and decrease in reduced by-products (1,3-propanediol and ethanol) formation. APH inclusion enhanced H2 production, and decreased substrate inhibition.

Toxicity of chlortetracycline and its metal complexes to model microorganisms in wastewater sludge.

Complexation of antibiotics with metals is a well-known phenomenon. Wastewater treatment plants contain metals and antibiotics, thus it is essential to know the effect of these complexes on toxicity towards microorganisms, typically present in secondary treatment processes. In this study, stability constants and toxicity of chlortetracycline (CTC) and metal (Ca, Mg, Cu and Cr) complexes were investigated. The calculated stability constants of CTC-metal complexes followed the order: Mg-CTC>Ca-CTC>Cu-CTC>Cr-CTC. Gram positive Bacillus thuringiensis (Bt) and Gram negative Enterobacter aerogenes (Ea) bacteria were used as model microorganisms to evaluate the toxicity of CTC and its metal complexes. CTC-metal complexes were more toxic than the CTC itself for Bt whereas for Ea, CTC and its metal complexes showed similar toxicity. In contrast, CTC spiked wastewater sludge (WWS) did not show any toxic effect compared to synthetic sewage. This study provides evidence that CTC and its metal complexes are toxic to bacteria when they are biologically available. As for WWS, CTC was adsorbed to solid part and was not biologically available to show measurable toxic effects.

Novel spectrophotometric method for detection and estimation of butanol in acetone-butanol-ethanol fermenter.

A new, simple, rapid and selective spectrophotometric method has been developed for detection and estimation of butanol in fermentation broth. The red colored compound, produced during reduction of diquat-dibromide-monohydrate with 2-mercaptoethanol in aqueous solution at high pH (>13), becomes purple on phase transfer to butanol and gives distinct absorption at λ520nm. Estimation of butanol in the fermentation broth has been performed by salting out extraction (SOE) using saturated K3PO4 solution at high pH (>13) followed by absorbance measurement using diquat reagent. Compatibility and optimization of diquat reagent concentration for detection and estimation of butanol concentration in the fermentation broth range was verified by central composite design. A standard curve was constructed to estimate butanol in acetone-ethanol-butanol (ABE) mixture under optimized conditions. The spectrophotometric results for butanol estimation, was found to have 87.5% concordance with the data from gas chromatographic analysis.

Genetic Engineering Strategies for Enhanced Biodiesel Production.

The focus on biodiesel research has shown a tremendous growth over the last few years. Several microbial and plant sources are being explored for the sustainable biodiesel production to replace the petroleum diesel. Conventional methods of biodiesel production have several limitations related to yield and quality, which led to development of new engineering strategies to improve the biodiesel production in plants, and microorganisms. Substantial progress in utilizing algae, yeast, and Escherichia coli for the renewable production of biodiesel feedstock via genetic engineering of fatty acid metabolic pathways has been reported in the past few years. However, in most of the cases, the successful commercialization of such engineering strategies for sustainable biodiesel production is yet to be seen. This paper systematically presents the drawbacks in the conventional methods for biodiesel production and an exhaustive review on the present status of research in genetic engineering strategies for production of biodiesel in plants, and microorganisms. Further, we summarize the technical challenges need to be tackled to make genetic engineering technology economically sustainable. Finally, the need and prospects of genetic engineering technology for the sustainable biodiesel production and the recommendations for the future research are discussed.

A novel anaerobic two-phase system for biohydrogen production and in situ extraction of organic acid byproducts.

Owing to CO2-free emission, hydrogen is considered as a potential green alternative of fossil fuels. Water is the major emission of hydrogen combustion process and gravimetric energy density of hydrogen is nearly three times more than that of gasoline and diesel fuel. Biological hydrogen production, therefore, has commercial significance; especially, when it is produced from low-cost industrial waste-based feedstock. Light independent anaerobic fermentation is simple and mostly studied method of biohydrogen production. During hydrogen production by this method, a range of organic acid byproducts are produced. Accumulation of these byproducts is inhibitory for hydrogen production as it may result in process termination due to sharp decrease in medium pH or by possible metabolic shift. For the first time, therefore, a two-phase anaerobic bioreactor system has been reported for biohydrogen production which involves in situ extraction of different organic acids. Among different solvents, based on biocompatibility oleyl alcohol has been chosen as the organic phase of the two-phase system. An organic:aqueous phase ratio of 1:50 has been found to be optimum for hydrogen production. The strategy was capable of increasing the hydrogen production from 1.48 to 11.65 mmol/L-medium.

Economic process to produce biohydrogen and volatile fatty acids by a mixed culture using vinasse from sugarcane ethanol industry as nutrient source.

This work evaluates the potential of vinasse (a waste obtained at the bottom of sugarcane ethanol distillation columns) as nutrient source for biohydrogen and volatile fatty acids production by means of anaerobic consortia. Two different media were proposed, using sugarcane juice or molasses as carbon source. The consortium LPBAH1 was selected for fermentation of vinasse supplemented with sugarcane juice, resulting in a higher H2 yield of 7.14 molH2 molsucrose(-1) and hydrogen content in biogas of approx. 31%, while consortium LPBAH2 resulted in 3.66 molH2/molsucrose and 32.7% hydrogen content in biogas. The proposed process showed a rational and economical use for vinasse, a mandatory byproduct of the renewable Brazilian energy matrix.

Mitigation of the inhibitory effect of soap by magnesium salt treatment of crude glycerol--a novel approach for enhanced biohydrogen production from the biodiesel industry waste.

Owing to its inhibitory effect on microbial growth, soap present in crude glycerol (CG) is a concern in biological valorization of the biodiesel manufacturing waste. By salting out strategy, up to 42% of the soap has been removed and the approach has beneficial effect on H2 production; however, removal of more than 7% of the soap was found to be inhibitory. Actually, soap is utilized as a co-substrate and due to removal; the carbon-nitrogen ratio of the medium might have decreased to reduce the production. Alternatively, without changing the carbon-nitrogen ratio of CG, MgSO4 treatment can convert the soap to its inactive form (scum). The approach was found to increase the H2 production rate (33.82%), cumulative H2 production (34.70%) as well as glycerol utilization (nearly 2.5-folds). Additionally, the treatment can increase the Mg (a nutrient) content of the medium from 0.57 ppm to 201.92 ppm.

Bio-hydrogen production by biodiesel-derived crude glycerol bioconversion: a techno-economic evaluation.

Global biodiesel production is continuously increasing and it is proportionally accompanied by a huge amount of crude glycerol (CG) as by-product. Due to its crude nature, CG has very less commercial interest; although its pure counterpart has different industrial applications. Alternatively, CG is a very good carbon source and can be used as a feedstock for fermentative hydrogen production. Further, a move of this kind has dual benefits, namely it offers a sustainable method for disposal of biodiesel manufacturing waste as well as produces biofuels and contributes in greenhouse gas (GHG) reduction. Two-stage fermentation, comprising dark and photo-fermentation is one of the most promising options available for bio-hydrogen production. In the present study, techno-economic feasibility of such a two-stage process has been evaluated. The analysis has been made based on the recent advances in fermentative hydrogen production using CG as a feedstock. The study has been carried out with special reference to North American biodiesel market; and more specifically, data available for Canadian province, Québec City have been used. Based on our techno-economic analysis, higher production cost was found to be the major bottleneck in commercial production of fermentative hydrogen. However, certain achievable alternative options for reduction of process cost have been identified. Further, the process was found to be capable in reducing GHG emissions. Bioconversion of 1 kg of crude glycerol (70 % w/v) was found to reduce 7.66 kg CO(2) eq (equivalent) GHG emission, and the process also offers additional environmental benefits.