Purification of Natural Pigments Violacein and Deoxyviolacein Produced by Fermentation Using Yarrowia lipolytica.

Violacein and deoxyviolacein are bis-indole pigments synthesized by a number of microorganisms. The present study describes the biosynthesis of a mixture of violacein and deoxyviolacein using a genetically modified Y. lipolytica strain as a production chassis, the subsequent extraction of the intracellular pigments, and ultimately their purification using column chromatography. The results show that the optimal separation between the pigments occurs using an ethyl acetate/cyclohexane mixture with different ratios, first 65:35 until both pigments were clearly visible and distinguishable, then 40:60 to create a noticeable separation between them and recover the deoxyviolacein, and finally 80:20, which allows the recovery of the violacein. The purified pigments were then analyzed by thin-layer chromatography and nuclear magnetic resonance.

Extraction of Valuable Biomolecules from the Microalga Haematococcus pluvialis Assisted by Electrotechnologies.

The freshwater microalga Haematococcus pluvialis is well known as the cell factory for natural astaxanthin, which composes up to 4-7% of its total dry weight. The bioaccumulation of astaxanthin in H. pluvialis cysts seems to be a very complex process that depends on different stress conditions during its cultivation. The red cysts of H. pluvialis develop thick and rigid cell walls under stress growing conditions. Thus, the biomolecule extraction requires general cell disruption technologies to reach a high recovery rate. This short review provides an analysis of the different steps in H. pluvialis's up and downstream processing including cultivation and harvesting of biomass, cell disruption, extraction and purification techniques. Useful information on the structure of H. pluvialis's cells, biomolecular composition and properties and the bioactivity of astaxanthin is collected. Special emphasis is given to the recent progress in application of different electrotechnologies during the growth stages and for assistance of the recovery of different biomolecules from H. pluvialis.

Use of instant controlled pressure drop technology for the preservation of edible insects' (yellow mealworm) quality: assessment of microbial inactivation.

The world population is expanding, and with it, so is the need for proteins for the food and feed sectors. Conventional livestock production is correlated with negative environmental repercussions such as global warming, land degradation, and biodiversity loss. The nutritional content of edible insects is comparable to that of conventional meat, and insect farming offers various environmental advantages over livestock production, making it a favorable sustainable protein resource. However, to be placed on the market, insects should be processed, and their microbial load should be within the accepted range of the European Commission regulation (EU) 2021/882. The purpose of this research is to investigate the use of instant controlled pressure drop (DIC) technology for the microbial inactivation of yellow mealworm larvae. This innovative decontamination treatment was compared to blanching in hot water, the conventional processing method. We also investigated the impacts on enzymatic browning. Results showed that the application of rapid (20 s) DIC treatment at 0.3 MPa allowed reducing the total aerobic count and the total yeast and mold count of larvae below the thresholds authorized by the commission regulation. Pressure, temperature, and number of cycle factors were found to have significant effects on the decontamination, while the treatment time had no effect for most treatments. In regard to blanching, we were able to get the authorized load at 90 °C for 360 s. Decimal reduction times for DIC and blanching treatments were found to be 3.8 s and 67.8 s for total aerobic count, and 3 s and 57 s for total yeast and mold count, respectively. Furthermore, DIC-treated larvae showed a significant increase in juice color brightness, which could be traced back to the oxidative enzymatic inactivation of larvae, whereas there was a slight difference between blanched and untreated larvae juice colors.

Impact of Electric Arcs and Pulsed Electric Fields on the Functional Properties of Beta-Lactoglobulin.

Beta-lactoglobulin (ß-lg) is a major whey protein with various techno-functional properties that can be improved by several treatments. Therefore, the objective of this study was to explore the impact of green high-voltage electrical treatments (HVETs)-namely, pulsed electric fields and electric arcs-on the functional properties of ß-lg. Both emulsifying and foaming stability and capacity, as well as the hygroscopicity of non-treated and pretreated ß-lg, were explored. The results demonstrated that the emulsifying capacity and stability of pretreated samples increased by 43% and 22% when compared to native ß-lg, respectively. Likewise, the pretreated ß-lg displayed better foaming stability compared to native ß-lg. In addition, the HVETs significantly decreased the hygroscopicity of ß-lg (by 48% on average), making it a good ingredient with reduced hygroscopicity for the food industry.

Mitigation of Acrylamide in Potato Chips by Pre-drying and Pulsed Electric Field Treatment.

The object of this work was to study the effects of preliminary vacuum drying (VD), pulsed electric field (PEF) treatment, frying temperature on color, oil uptake, and acrylamide (AA) content in fried potato chips. The results of this study indicated that an increase of frying temperature from 120 to 180°C led to a decrease of frying time of around 70% for untreated and PEF pre-treated samples. The color value of L* and a* decreased with the increase of frying temperature, and those values of the sample pre-treated by PEF were significantly higher compared to those obtained from untreated samples. The PEF pre-treatment promoted the reduction of oil content of fried samples by up to 17.6, 14.2, and 16% compared with untreated samples at the frying temperatures of 120, 150, and 180°C, respectively. Higher efficiency was observed by applying the preliminary VD in the case of the frying temperature of 150°C. Furthermore, it was revealed that PEF pre-treatment and preliminary VD application lead to a synergetic effect on the reduction of AA content in potato chips. For example, with the initial moisture ratio of 0.5, pre-dried by VD and pre-treated by PEF, the AA content was noticeably decreased from 2,220 to 311 µg/kg compared to untreated and undehydrated samples at the frying temperature of 150°C. Our findings provide reference for a new pre-treatment to mitigate AA formation and to improve the quality of potato chips.

Mechanical damage and thermal effect induced by ultrasonic treatment in olive leaf tissue. Impact on polyphenols recovery.

The influence of ultrasound treatment (US) on cellular damage of olive leaf tissue was studied. Mechanical damage and thermal effect of US were characterized. The level of tissue damage was defined by the diffusivity disintegration index ZD based on the diffusivity of solutes extracted from olive leaves differently treated. The Arrhenius form using the temperature dependences of the thermal treatment time within the temperature interval 20-90 °C was observed for the thermal process. The corresponding activation energy ΔUT was estimated as 57 kJ/mol. The temperature dependences of electrical conductivity were measured for extracts of intact and maximally treated olive leaves. Then the diffusivity disintegration index ZD and total phenolic compounds recovery for three studied US powers were calculated (100, 200, and 400 W). The results evidenced that the mechanically stimulated damage in olive leaf tissue can occur even at a low US power of 100 W if treatment time is long enough (t = 3.5 h). The US treatment noticeably accelerated the diffusion process mechanically in addition to its thermal effect. Trials in aqueous solution revealed the dependence of polyphenols extraction on damage level with respect to the US power applied.

Monte Carlo simulation of dead-end diafiltration of bidispersed particle suspensions.

A two-dimensional Monte Carlo simulation of dead-end diafiltration of bidispersed particle suspensions was performed. The diafiltration process involves separation of components based on their size by using a permeable membrane. The continuous model was applied to study separation of mixture of disks with diameter d and D (D>d). It was assumed that the membrane at the bottom was permeable to the particles of the smaller diameter d, and impermeable to the particles of the larger diameter D. The process of vertical filtration was accompanied by the simultaneous Brownian motion of the particles and downward movement of the piston. The mixtures with different numerical concentrations of particles, diameter ratio, and initial size of the systems in the vertical direction L_{y} have been studied. The time dependencies of the rejection coefficient k and relative height of suspension h/L_{y} revealed the presence of complete and incomplete separation regimes. The presence of filtration and diffusion-driven stratification of the disks in the vertical direction was observed. The phenomenon of incomplete separation was explained by the formation of an impenetrable barrier from larger particles at the bottom of the deposit.

Recent insights in the impact of emerging technologies on lactic acid bacteria: A review.

Lactic acid bacteria (LAB) have a long history of applications in the food industry for fermentation and preservation. This feature is due to their metabolic products that can improve the nutritional and sensory characteristics of foods as well as their antimicrobial compounds that contribute to extend the shelf life of food products. Some emerging technologies including pulsed electric fields (PEF), power ultrasound (US), high-pressure processing (HPP), ultraviolet (UV), and microwave (MW) have attracted great attention for their implementation in the food industry as mild processing technologies. They have the advantage of efficiently inactivating the microorganisms, along with maintaining the fresh attributes of the food products. When applied at a sub-lethal level, these technologies present the potential to enhance several processes, such as improved microbial growth and fermentation conditions, as well as modified metabolic properties of LAB. This review covers the characteristics of LAB and their applications in the food industry. It discusses the impacts of emerging technologies on these microorganisms, with a special focus on microbial inactivation, growth stimulation, and improvement of the beneficial features of LAB by emerging technologies.

Impact of pulsed electric fields on vacuum drying kinetics and physicochemical properties of carrot.

The effect of pulsed electric fields (PEF) on the kinetics of vacuum drying (VD) and rehydration of carrot tissue was studied. The intensity of PEF treatment was E = 0.6 kV/cm and the total PEF treatment duration was tPEF = 0.1 s. The VD was performed at sub-atmospheric pressure p = 3.03 × 104 Pa for different temperatures Td = 25 °C and 90 °C. During VD, the kinetics of moisture removal and temperature evolution inside of untreated and PEF pretreated samples were studied. The color and structure of the samples were also investigated. PEF pretreatment enhanced VD kinetics at different studied temperatures. However, its effect on VD was more evident at lower drying temperatures. With the PEF pretreatment, the drying time was reduced by 55% and 33% at 25 °C and 90 °C respectively. After the drying and rehydration process, the change in color of PEF pretreated samples was less significant compared to untreated ones. After rehydration, the untreated and PEF pretreated samples were nearly restored to their initial form and size.

Impact of the Physicochemical Composition and Microbial Diversity in Apple Juice Fermentation Process: A Review.

Fermented apple beverages are produced all over the world with diverse characteristics associated with each country. Despite the diversifications, cider producers are confronted with similar issues and risks. The nature of the raw material, also known as the fermentation medium, plays a key role in fermentation. A well-defined composition of apples is, therefore, required to produce cider with good quality. In addition, ferment and its metabolism are important factors in the fermentation process. The producers of cider and other alcoholic beverages are looking in general for novel yeast strains or for the use of native strains to produce "authentic" and diversified beverages that are distinct from each other, and that attract more and more consumers. Research articles on cider production are infrequent compared to wine production, especially on the impact of the chemical composition and microbial diversity of apples on fermentation. Even though the processing of fermented beverages is close in terms of microbial interactions and production, the study of the specific properties of apples and the production challenges of cider production is advantageous and meaningful for cider producers. This review summarizes the current knowledge on apple composition and the impact of the must composition on fermentation and yeast growth. In addition, the microbial diversity of cider, activities, and its influence on fermentation are reviewed.

Effect of Pulsed Electric Fields on the Growth and Acidification Kinetics of Lactobacillus delbrueckii Subsp. bulgaricus.

The aim of this work was to investigate the effect of pulsed electric fields (PEF) on the growth and acidification kinetics of Lactobacillus delbrueckii subsp. bulgaricus CFL1 during fermentation. The PEF treatments were applied during the fermentation process using a recirculation pump and a PEF treatment chamber coupled with a PEF generator. The medium flow rate through the chamber was first optimized to obtain the same growth and acidification kinetics than the control fermentation without medium recirculation. Different PEF intensities (60-428 V cm-1) were then applied to the culture medium to study the impact of PEF on the cells' behavior. The growth and acidification kinetics were recorded during the fermentation and the specific growth rates µ, pH, and acidification rate (dpH/dt) were assessed. The results obtained showed a biphasic growth by applying high PEF intensities (beyond 285 V cm-1) with the presence of two maximal specific growth rates and a decrease in the acidification activities. It was demonstrated that the cells were stressed during the PEF treatment, but presented an accelerated growth after stopping it, leading thereby to similar absorbance and pH at the end of the fermentation. These results show the great potential of PEF technology to be applied to generate low acidified products by performing PEF-assisted fermentations.

Evaluation of the fermentative capacity of an indigenous Hanseniaspora sp. strain isolated from Lebanese apples for cider production.

The present work studied the fermentative potential and carbon metabolism of an indigenous yeast isolated from Lebanese apples for cider production. The indigenous yeast strain was isolated from a spontaneous fermented juice of the Lebanese apple variety 'Ace spur'. The sequencing of the Internal Transcribed Spacer (ITS) domain of rRNA identified the isolated yeast strain as a member of the Hanseniaspora genus. These results suggest an intragenomic ITS sequence heterogeneity in the isolated yeast strain specifically in its ITS1 domain. The different investigations on the yeast carbon metabolism revealed that the isolated yeast is 'Crabtree positive' and can produce and accumulate ethanol from the first hours of fermentation. Thus, our findings highlight the possibility of using the isolated indigenous Hanseniaspora strain as a sole fermentative agent during cider production.

Pulsed electric field-assisted fermentation of Hanseniaspora sp. yeast isolated from Lebanese apples.

Hanseniaspora sp. yeast was stimulated using pulsed electric field (PEF) during the different fermentation phases. The impact of PEF parameters on the growth rate and substrate consumption was studied. The PEF intensities chosen for this study were mainly in the range of 72-285 V cm-1. A PEF treatment chamber was designed for this study with a ratio of 1:50 between the volume of the fermenter and the volume of the chamber. It allows the recycling of the culture medium using a peristaltic pump, and the yeast treatment by PEF during the fermentation. The continuous circulation of the medium allows avoiding the increase of the temperature inside the fermenter, the cell aggregation, as well as the agitation and the scale-up issues that are associated with the PEF treatment of the entire volume in batch mode. The maximal yeast growth rate was obtained using an electric field strength of 285 V cm-1 applied during both Lag and early exponential phase, and Log phase. This observation was accompanied by a faster consumption of glucose in the medium during the fermentation. Besides, the sensitivity of Hanseniaspora sp. yeast to PEF treatment was more pronounced during the Lag and early exponential phase than the Log phase. The results obtained exposed the great benefit of stimulating Hanseniaspora sp. yeast using moderate PEF as it reduces the fermentation time along with increasing the biomass concentration.

Two-step procedure for selective recovery of bio-molecules from microalga Nannochloropsis oculata assisted by high voltage electrical discharges.

Two-step procedure with the initial aqueous extraction from raw microalgae Nannochloropsis oculata and secondary organic solvent extraction from vacuum dried (VD) microalgae were applied for selective recovery of bio-molecules. The effects of preliminary aqueous washing and high voltage electrical discharges (HVED, 40 kV/cm, 4 ms pulses) were tested. The positive effects of HVED treatment and washing on selectivity of aqueous extraction of ionics and other water-soluble compounds (carbohydrates, proteins and pigments) were observed. Moreover, the HVED treatment allowed improving the kinetic of vacuum drying, and significant effects of HVED treatment on organic solvent extraction of chlorophylls, carotenoids and lipids were determined. The proposed two-step procedure combining the preliminary washing, HVED treatment and aqueous/organic solvents extraction steps are useful for selective extraction of different bio-molecules from microalgae biomass.

Ultrasound-assisted fermentation for cider production from Lebanese apples.

The present work studies the impact of low-intensity ultrasound (US) on Hanseniaspora sp. yeast fermentations. The effect of pulse duration and growth phase on US application was first evaluated using a synthetic medium. The optimal conditions were then applied to apple juice US-assisted fermentation. An US treatment chamber was first designed to allow the recycling of the culture medium. The optimal US pulse duration on the yeast growth rate was of 0.5 s followed by 6 s rest period, and during 6 h of both Lag and Log phases. These US parameters led to a faster consumption of glucose in the medium during the fermentation, compared to the untreated culture. The impact of US was also depending on the growth phase, showing higher sensitivity of the yeast to US during the Lag phase rather than the Log phase. US-assisted fermentation of apple juice showed a significant increase in biomass growth and glucose consumption, along with a significant decrease in the ethanol yield. The fastest growth kinetic (by 52%), and the highest ethanol reduction (by 0.55% (v, v)) were obtained for the treatment during the first 12 h of fermentation, thereby, the stationary phase was reached faster, and the maximum biomass growth rate was 10 folds higher compared to the untreated culture. The results obtained in this study demonstrated the promising efficiency of US-assisted fermentation in stimulating the biomass growth and reducing the ethanol content in alcoholic beverages.

Green extraction of polyphenols from grapefruit peels using high voltage electrical discharges, deep eutectic solvents and aqueous glycerol.

Deep eutectic solvents (DES) and aqueous glycerol were proposed as green alternatives to conventional solvents for the extraction of polyphenols from grapefruit peels. In order to increase the extraction kinetics and yields of polyphenols, high voltage electrical discharges (HVED) were used as a pre-treatment technology (energy varied between 7.27 and 218 kJ/kg). Results showed that the HVED energy input can be reduced, when the subsequent solid-liquid extraction was performed in 20% (w/v) aqueous glycerol or in DES (lactic acid: glucose) instead of water. The addition of glycerol has reduced the energy of the pre-treatment by 6 times. The same diffusivity of polyphenols (4 × 10-11 m2/s) was obtained in water from HVED pre-treated peels at 218 kJ/kg and in aqueous glycerol from pre-treated peels at 36 kJ/kg. The solubility of naringin, the main flavonoid compound of grapefruit peels in the solvents, was investigated through a theoretical modelling of its Hansen solubility parameters.

Ultrasound assisted purification of polyphenols of apple skins by adsorption/desorption procedure.

The ultrasound (US) assisted purification of polyphenols of apple skins by adsorption/desorption on the poly-aromatic Amberlite adsorbent XAD-16 was studied. The adsorption steps were done at different temperatures (T = 25-40 °C) with application of US at different intensities (P = 0-400 W). The desorption steps were tested in aqueous ethanol solution at different concentrations of ethanol (Cet = 0-96%). The isotherm of polyphenol adsorption was well described using the Freundlich model. The data on adsorption kinetics and static isotherm evidenced the presence of adsorption on heterogeneous surface with broad distribution of adsorption times that can depend on content of polyphenols in the solutions and applied power of sonication. The studies of desorption revealed the optimum desorption efficiency of polyphenols at 50% concentration of ethanol. The desorption ratio was positively affected by the sonication during the adsorption step. The highest adsorption/desorption efficiency (recovery) was observed for polyphenols as compared with proteins and soluble matter content and it reached of ≈30.6% (0 W) and 68.9% (50 W) in absence and presence of sonication, respectively. The effects of high US power on the damage of XAD-16 were discussed. The obtained data evidenced on good perspective of application of adsorption/desorption procedure assisted by sonication for purification of polyphenols from apple skin extracts.

Effects of Pulsed Electric Field Treatment on Compression Properties and Solutes Diffusion Behaviors of Jerusalem artichoke.

Jerusalem artichoke is widely used as raw material for industrial production of inulin. Pressing (compression) and diffusion are two effective technologies for bio-compounds' recovery from plants. In this work, pulsed electric field (PEF) treatment at 400, 600, and 800 V/cm during 100 ms was applied to facilitate juice and solutes recovery from Jerusalem artichoke. The application of PEF led to electroporation of cell membranes and enhanced the tissue compression/juice expression and solutes diffusion. The consolidation coefficient (calculated by application of semi-empirical model) of PEF treated sample at 800 V/cm was 6.50 × 10-7 m²/s, which is significantly higher than that of untreated sample (5.02 × 10-9 m²/s) and close to that of freeze-thawed sample. Diffusion experiments with PEF treated samples were carried out at 25, 50, and 75 °C. A PEF treatment of Jerusalem artichoke at 800 V/cm led to a similar diffusion behavior at 25 °C, compared to diffusion behavior obtained from untreated sample at 75 °C.

Application of high-voltage electrical discharges and high-pressure homogenization for recovery of intracellular compounds from microalgae Parachlorella kessleri.

Treatments with high-voltage electrical discharges (HVED) and high-pressure homogenization (HPH) were studied and compared for the release of ionic components, carbohydrates, proteins, and pigments from microalgae Parachlorella kessleri (P. kessleri). Suspensions (1% w/w) of microalgae were treated by HVED (40 kV/cm, 1-8 ms) or by HPH (400-1200 bar, 1-10 passes). Particle-size distribution (PSD) and microscopic analyses were used to detect the disruption and damage of cells. HVED were very effective for the extraction of ionic cell components and carbohydrates (421 mg/L after 8 ms of the treatment). However, HVED were ineffective for pigments and protein extraction. The concentration of proteins extracted by HVED was just 750 mg/L and did not exceed 15% of the total quantity of proteins. HPH permitted an effective release overall of intracellular compounds from P. kessleri microalgae including a large quantity of proteins, whose release (at 1200 bar) was 4.9 times higher than that obtained by HVED. Consequently, HVED can be used at the first step of the overall extraction process for the selective recovery of low-molecular-weight components. HPH can be then used at the second step for the recovery of remaining cell compounds.