Deep Eutectic Solvents for Biodiesel Purification in a Microextractor: Solvent Preparation, Selection and Process Optimization.

The most important and commonly used process for biodiesel synthesis is transesterification. The main by-product of biodiesel synthesis by transesterification is glycerol, which must be removed from the final product. Recently, deep eutectic solvent (DES) assisted extraction has been shown to be an effective and sustainable method for biodiesel purification. In this study, biodiesel was produced by lipase-catalysed transesterification from sunflower oil and methanol. A total of 12 different eutectic solvents were prepared and their physical properties were determined. Mathematical models were used to define which physical and chemical properties of DES and to what extent affect the efficiency of extraction of glycerol from the biodiesel. After initial screening, cholinium-based DES with ethylene glycol as hydrogen bond donor was selected and used for optimization of extraction process conditions performed in a microsystem. To determine the optimal process conditions (temperature, biodiesel:DES volume ratio, residence time), the experimental three-level-three-factor Box-Behnken experimental design was used. In the end, a combination of a mathematical model and experimental results was used to estimate how many micro-extractors are necessary for the complete removal of glycerol.

Changes in the content of glucosinolates, polyphenols and carotenoids during lactic-acid fermentation of cruciferous vegetables: A mini review.

Cruciferous vegetables are considered functional foods because of their content of health-related compounds. They are grown and consumed in various cultures around the world. Fermentation as a preservation method for cruciferous vegetables has been used since ancient times. This process results in fermented products that have a unique flavour and odour, high bioactivity, and a distinctly different phytochemical profile than raw vegetables. In this mini review, we summarize data on changes in phytochemical content during lactic-acid fermentation of various cruciferous vegetables. The main focus was on the changes in the group of glucosinolates, polyphenols and carotenoids.

Development of NIR-Based ANN Models for On-Line Monitoring of Glycerol Concentration during Biodiesel Production in a Microreactor.

During the production process, a whole range of analytical methods must be developed to monitor the quality of production and the desired product(s). Most of those methods belong to the group of off-line monitoring methods and are usually recognized as costly and long-term. In contrast, on-line monitoring methods are fast, reliable, simple, and repeatable. The main objective of this study was to compare different methods for monitoring total glycerol concentration as one of the indicators of process efficiency during biodiesel production in a batch reactor and in a microreactor. During the biodiesel production process, the glycerol concentration was measured off-line using standard methods based on UV-VIS spectrophotometry and gas chromatography. Neither method provided satisfactory results, namely, both analyses showed significant deviations from the theoretical value of glycerol concentration. Therefore, near infrared spectroscopy (NIR) analysis was performed as an alternative analytical method. The analysis using NIR spectroscopy was performed in two ways: off-line, using a sample collected during the transesterification process, and on-line by the continuous measurement of glycerol concentration in a rector. Obtained results showed a great NIR application potential not only for off-line but also for on-line monitoring of the biodiesel production process.

An enhanced composting process with bioaugmentation: Mathematical modelling and process optimization.

In this paper, two different types of biowaste composting processes were carried out - composting without and with bioaugmentation. All experiments were performed in an adiabatic reactor for 14 days. Composting enhanced with bioaugmentation was the better choice because the thermophilic phase was achieved earlier, making the composting time shorter. Additionally, a higher conversion of substrate (amount of substrate consumed) was also noticed in the process enhanced by bioaugmentation. A mathematical model was developed and process parameters were estimated in order to optimize the composting process. Based on good agreement between experimental data and the mathematical model simulation results, a three-level-four-factor Box-Behnken experimental design was employed to define the optimal process conditions for further studies. It was found that the air flow rate and the mass fraction of the substrate have the most significant effect on the composting process. An improvement of the composting process was achieved after altering the mentioned variables, resulting in shorter composting time and higher conversion of the substrate.

Transesterification in Microreactors-Overstepping Obstacles and Shifting Towards Biodiesel Production on a Microscale.

Biodiesel, which was earlier used only as an alternative fuel, is now an indispensable component of commercial diesel. Conventional production processes are unable to cope with the increasing demand for biodiesel, and therefore more and more work is being done to intensify the existing processes. The intensification of the biodiesel production process, taking into account the environmental and economic factors, is based on increasing productivity. One way to achieve that is by reducing the volume of production units. The application of the enzymatic reaction path, while reducing the volume of process equipment to the micro-level, has significantly magnified the productivity of the biodiesel production process, which is primarily due to better mass transfer in microsystems. Additional breakthrough is the use of deep eutectic solvents (DES) instead of buffers for enzyme stabilization. In this study, a lipase from Thermomyces lanuginosus (TlL) (both commercial and produced by solid-state fermentation) was used as a catalyst for biodiesel production. Edible and waste sunflower oil, as well as methanol, were used as substrates. The reaction mediums were buffer and DES. The transesterification reaction was carried out in a batch reactor and the emphasis was made on different microreactor configurations. The highest yield of 32% for residence time of only τ = 30 min was obtained in the microreactor system with an emulsion of waste oil and a commercial enzyme suspended in a buffer. This indicates that enzymatic transesterification could be a valuable reaction path for dealing with waste oils. Furthermore, biodiesel synthesis in DES showed somewhat lower yields, but by increasing the water content in the system, the reaction could prove much better results. In the end, the effects of reaction conditions on the volumetric productivity of the process were analyzed.

Kinetic Parameter Estimation and Mathematical Modelling of Lipase Catalysed Biodiesel Synthesis in a Microreactor.

Development of green, clean, and sustainable processes presents new challenges in today's science. Production of fuel is no exception. Considering the utilisation of various renewable sources, the synthesis of biodiesel, characterised as more environmentally-friendly then fossil fuel, has drawn significant attention. Even though the process based on chemical transesterification in a batch reactor still presents the most used method for its production, enzyme catalysed synthesis of biodiesel in a microreactor could be a new approach for going green. In this research, edible sunflower oil and methanol were used as substrates and lipase from Thermomyces lanuginosus (Lipolase L100) was used as catalyst for biodiesel synthesis. Experiments were performed in a polytetrafluoroethylene (PTFE) microreactor with three inlets and in glass microreactors with two and three inlets. For a residence time of 32 min, the fatty acids methyl esters (FAME) yield was 30% higher than the yield obtained for the glass microreactor with three inlets. In comparison, when the reaction was performed in a batch reactor (V = 500 mL), the same FAME yield was achieved after 1.5 h. In order to enhance the productivity of the process, we used proposed reaction kinetics, estimated kinetic parameters, and a mathematical model we developed. After validation using independent experimental data, a proposed model was used for process optimization in order to obtain the highest FAME yield for the shortest residence time.

Lipase catalysed biodiesel synthesis with integrated glycerol separation in continuously operated microchips connected in series.

Although the application of microreactors in different processes has been extensively explored in recent decades, microreactors continue to be underexplored in the context of the enzyme-catalysed process for biodiesel production. Due to their numerous advantages, microreactors could become the next step in the development of a biodiesel production process characterised by sustainability, cost-effectiveness and energy efficiency. In this investigation, biodiesel production was catalysed by lipase from Thermomyces lanuginosus (Lipolase L100). Edible sunflower oil was used as a model substrate in order to investigate the process. After optimal process conditions had been determined, waste-cooking oil was used for biodiesel production to make the production process more sustainable. Three different substrate-feeding strategies were investigated and finally an optimal strategy was proposed. In all the investigated systems, fatty acids methyl esters (FAME) content was higher than 95% and obtained in a significantly shorter time (less than 2 h) compared to the batch process in which biodiesel production was catalysed by lipase (C = 95%, t = 96 h). After the optimal biodiesel production system had been proposed, an integrated system with two microchips connected in series was developed. The first microchip was used for biodiesel production and the second for simultaneous purification i.e. glycerol separation. Finally, purified biodiesel was produced with glycerol content below the detection limit.

Synergy of Microtechnology and Biotechnology: Microreactors as an Effective Toolfor Biotransformation Processes§: §The paper was presented at European Biotechnology Congress, 25-27 May 2017, Dubrovnik, Croatia.

Despite the fact that microreactors have been present for more than 40 years now and that their potential has been extensively exploited in chemical synthesis, analytics and screening, to date very few biocatalytic processes have been explored in microreactors. It is claimed that enzymatic microreactor technology is exactly in the same place where chemical microreactors were 15 years ago. However, general opinion is that the efforts devoted to the research of micro-enzymatic reactors will inaugurate a new breakthrough in bio-based processing. The aim of this review is to explore the synergy between microtechnology, mainly microreactors, and biotechnology, and to assess its potential, opportunities, challenges and future application in biotechnology.

Catechol Removal from Aqueous Media Using Laccase Immobilized in Different Macro- and Microreactor Systems.

Laccase belongs to the group of enzymes that are capable to catalyze the oxidation of phenols. Since the water is only by-product in laccase-catalyzed phenol oxidations, it is ideally "green" enzyme with many possible applications in different industrial processes. To make the oxidation process more sustainable in terms of biocatalyst consumption, immobilization of the enzyme is implemented in to the processes. Additionally, when developing a process, choice of a reactor type plays a significant role in the total outcome.In this study, the use of immobilized laccase from Trametes versicolor for biocatalytic catechol oxidation was explored. Two different methods of immobilization were performed and compared using five different reactor types. In order to compare different systems used for catechol oxidation, biocatalyst turnover number and turnover frequency were calculated. With low consumption of the enzyme and good efficiency, obtained results go in favor of microreactors with enzyme covalently immobilized on the microchannel surface.

Bioproduction of food additives hexanal and hexanoic acid in a microreactor.

Hexanal and hexanoic acid have number of applications in food and cosmetic industry because of their organoleptic characteristics. Problems like low yields, formation of unwanted by-products, and large quantities of waste in their traditional production processes are the reasons for developing new production methods. Biotransformation in a microreactor, as an alternative to classical synthesis processes, is being investigated. Because conditions in microreactors can be precisely controlled, the quality of the product and its purity can also be improved. Biocatalytic oxidation of hexanol to hexanal and hexanoic acid using suspended and immobilized permeabilized whole baker's yeast cells and suspended and immobilized purified alcohol dehydrogenase (ADH) was investigated in this study. Three different methods for covalent immobilization of biocatalyst were analyzed, and the best method for biocatalyst attachment on microchannel wall was used in the production of hexanal and hexanoic acid.