Patrick Bourseau

Impact of ultrafiltration and nanofiltration of an industrial fish protein hydrolysate on its bioactive properties

BACKGROUND Numerous studies have demonstrated that in vitro controlled enzymatic hydrolysis of fish and shellfish proteins leads to bioactive peptides. Ultrafiltration (UF) and/or nanofiltration (NF) can be used to refine hydrolysates and also to fractionate them in order to obtain a peptide population enriched in selected sizes. This study was designed to highlight the impact of controlled UF and NF on the stability of biological activities of an industrial fish protein hydrolysate (FPH) and to understand whether fractionation could improve its content in bioactive peptides. RESULTS The starting fish protein hydrolysate exhibited a balanced amino acid composition, a reproducible molecular weight (MW) profile, and a low sodium chloride content, allowing the study of its biological activity. Successive fractionation on UF and NF membranes allowed concentration of peptides of selected sizes, without, however, carrying out sharp separations, some MW classes being found in several fractions. Peptides containing Pro, Hyp, Asp and Glu were concentrated in the UF and NF retentates compared to the unfractionated hydrolysate and UF permeate, respectively. Gastrin/cholecystokinin-like peptides were present in the starting FPH, UF and NF fractions, but fractionation did not increase their concentration. In contrast, quantification of calcitonin gene-related peptide (CGRP)-like peptides demonstrated an increase in CGRP-like activities in the UF permeate, relative to the starting FPH. The starting hydrolysate also showed a potent antioxidant and radical scavenging activity, and a moderate angiotensin-converting enzyme (ACE)-1 inhibitory activity, which were not increased by UF and NF fractionation. CONCLUSION Fractionation of an FPH using membrane separation, with a molecular weight cut-off adapted to the peptide composition, may provide an effective means to concentrate CGRP-like peptides and peptides enriched in selected amino acids. The peptide size distribution observed after UF and NF fractionation demonstrates that it is misleading to characterize the fractions obtained by membrane filtration according to the MW cut-off of the membrane only, as is currently done in the literature.

Concentration and desalination of fish gelatin by ultrafiltration and continuous diafiltration processes

Abstract Ultrafiltration (UF) of gelatin liquor from marine source (tuna skins) has been performed on a tubular mineral membrane of cut-off 10,000 Da. An attempt was made to concentrate the fish gelatin as well as to purify it by means of the diafiltration technique (DF). Primary experiments were devoted to optimise the UF performances as a function of the main adjustable parameters. The influence of pH, transmembrane pressure, recirculating velocity, and working temperature on both volumetric permeate fluxes and protein retention rates was successively examined. Under the appropriate operating conditions, it was shown that the used membrane can achieve the initial concentration step of the gelatin solutions up to 12 wt% with a protein yield as high as 0.98. In addition, continuous DF proved useful to reduce the salt content of the solutions at an average desalting speed of 185 g/h.m2 with a few loss of protein in the permeate side.

Recovery by nanofiltration and reverse osmosis of marine flavours from seafood cooking waters

Abstract Cooking waters from buckies, shrimps and tuna have a high level of polluting load (Chemical Oxygen Demand — COD — comprised between 5 and 40 gO2/L) and have to be treated before being rejected in the environment. However, these juices seem to contain interesting flavour compounds. This recovery would allow the industrialists to diminish the waste water treatment cost and to recover high added value molecules. In this work, a membrane process system consisting in ultrafiltration (UF) followed by uanofiltration (NF) or reverse osmosis (RO) was used to reduce the pollution load and to concentrate flavour compounds of seafood cooking juices (buckies, shrimps and tuna). The first ultrafiltration step (cut-off: 0.02 μm) consisted in clarifying the cooking waters in order to increase fluxes of the following filtrations. NF (cut-off: 300 Da) did not seem to be efficient enough for flavour recovery, whereas RO was, for shrimps and buckies cooking juices. At the opposite, aromas contained in the tuna cooking juice were not perfectly retained by the RO membrane used. Nevertheless, the polluting load was considerably decreased after RO (COD reduced by 95% for shrimps and buckies and 85% for tuna).

Concentration polarization phenomenon during the nanofiltration of multi-ionic solutions: influence of the filtrated solution and operating conditions.

One of the major difficulties for the prediction of separation performances in the case of multi-ionic mixtures nanofiltration lies in the description of the concentration polarization phenomenon. Usual models available in literature do not take account of the polarization phenomenon or only describe it cursorily. Very few studies dedicated to the understanding and the specific description of the concentration polarization phenomenon are available in literature and a 2-D multi-ionic model describing the layer heterogeneity along the membrane length has never been proposed yet. The model used in the present work, called Pore and Polarization Transport Model (PPTM), allows an accurate description of the concentration polarization layer occurring during the filtration of multi-ionic solutions by taking account of the radial electromigrative transport in the layer, the turbulence, as well as the axial heterogeneity. In this context, the present paper aims at proposing a numerical investigation of the influence of operating conditions on the behavior of the polarization layer occurring at the membrane vicinity. The input parameters governing the transport through the membrane have been assessed in a previous study in the same experimental conditions so that only the polarization layer is investigated here. The proposed model which was previously validated on experimental observed rejection curves is then used to understand how operating conditions, such as applied pressure, feed flow-rate, or divalent ion proportion, govern the polarization phenomenon. For this purpose, concentration and thickness axial profiles along the membrane length and radial profiles within the polarization layer are investigated for various conditions. Finally, the impact of the type of divalent ion and the number of ions is also studied on various mixtures.

Application of response surface methodology to optimise the antioxidant activity of a saithe (Pollachius virens) hydrolysate.

The objective of this study was to produce, by an enzymatic hydrolysis process at a pilot scale, a saithe (Pollachius virens) hydrolysate with a high antioxidant activity. Design of experiment methodology, based on laboratory-scale experiments, was used to obtain a behavioral reduced model that allows one to determine the optimal operating conditions maximizing the antioxidant activity. Two specifications were studied: the degree of hydrolysis and the antioxidant activity. The effects of the following hydrolysis parameters (temperature, pH, enzyme concentration, and operating time) were studied and presented as response surfaces. From these results, a multifactorial optimization was performed and the Pareto optimal set of efficient solutions was evaluated. The optimal conditions were tested at laboratory scale and then validated by comparison with tests carried out on a pilot plant.

Optimization of bead milling parameters for the cell disruption of microalgae: process modeling and application to Porphyridium cruentum and Nannochloropsis oculata.

A study of cell disruption by bead milling for two microalgae, Nannochloropsis oculata and Porphyridium cruentum, was performed. Strains robustness was quantified by high-pressure disruption assays. The hydrodynamics in the bead mill grinding chamber was studied by Residence Time Distribution modeling. Operating parameters effects were analyzed and modeled in terms of stress intensities and stress number. RTD corresponded to a 2 CSTR in series model. First order kinetics cell disruption was modeled in consequence. Continuous bead milling was efficient for both strains disruption. SI-SN modeling was successfully adapted to microalgae. As predicted by high pressure assays, N. oculata was more resistant than P. cruentum. The critical stress intensity was twice more important for N. oculata than for P. cruentum. SI-SN modeling allows the determination of operating parameters minimizing energy consumption and gives a scalable approach to develop and optimize microalgal disruption by bead milling.

Harvesting Chlorella vulgaris by natural increase in pH: effect of medium composition

The freshwater microalga Chlorella vulgaris was harvested by autoflocculation resulting from the precipitation of magnesium or calcium compounds induced by a slow increase in pH in the absence of CO2 input. Autoflocculation was tested in two culture media with, respectively, ammonium (NH 4+) and nitrate (NO 3−) ions as nitrogen source. The culture pH increased because of photosynthesis and CO2 stripping. pH rose to 11 after 8 h in the NO 3− medium, but did not exceed 9 in the NH 4+ medium. No flocculation took place in any of the media. Autoflocculation tests were repeated in the NO 3− -based culture medium by progressively increasing the concentrations of Ca2+ and Mg2+ until inorganic compounds precipitated and flocculated microalgae. The minimal concentrations for flocculation were found to be 120 mg Ca2 L−1 and 1000 mg Mg2+ L−1. These values were, respectively, 3.5 times and 20 times higher than those allowing flocculation by NaOH addition. Energy-dispersive X-ray spectroscopy, zeta potential measurement, and ionic chromatography suggest that the mechanisms involved are different. The rate of cell removal was close to 90% in both cases, but cells were more concentrated in the aggregates obtained by magnesium compound precipitation, with an estimated concentration close to 33 g (dry matter) L−1, against 19 g L−1 for calcium phosphates.

Numerical Ways to Characterize the Deterioration of Nanofiltration Membranes

In this study, a transport model is used to characterize structural and physico-chemical changes in a nanofiltration membrane during the filtration of ionic mixtures. The membrane state is analyzed by a set of four model parameters identified from glucose and salts filtration: the membrane water permeability (Lp), the mean pore radius (rp), the membrane charge density (Xd), and the dielectric constant of the solution inside pores ( p). The study of these structural and physico-chemical properties allows us to determine if deterioration or fouling occurred during filtration. Two distinct identification procedures from filtration of synthetic solutions are investigated in this paper. One is based on the filtration of single salt solutions, whereas the other lies in parameters identification from mixtures containing at least three ions. These methods are applied here to characterize influence of fouling deposit formation and membrane cleaning.

Filterability of exopolysaccharides solutions from the red microalga Porphyridium cruentum by tangential filtration on a polymeric membrane

ABSTRACT The red microalga Porphyridium cruentum is exploited industrially for its exopolysaccharides (EPS) and pigments production. EPS produced by P. cruentum are partially released and dissolved into the surrounding environment, they can be recovered from the culture medium after removing the cells. This paper presents a parametric study of the ultrafiltration of EPS solutions on organic membrane. The EPS solutions were produced in conditions representative of an industrial production. They were filtered at lab-scale on a flat, PES 50 kDa MWCO membrane in a complete recirculation mode of permeate and retentate. Permeate flux-transmembrane pressure (TMP) curves were established up to the limiting flux for the filtration of solutions with various values of concentration in EPS (0.10–1.06 kg GlcEq m–3), fluid tangential velocity (0.3–1.2 m s–1) and temperature (20°C and 40°C). The reversible and irreversible parts of fouling were evaluated for each experiment and the critical flux was determined for an intermediate EPS concentration (0.16 kg GlcEq m–3). The results showed that EPS solutions had a strong fouling capacity. When filtering the lowest concentrated solution (0.10 kg GlcEq m–3) with moderate fouling conditions, the overall fouling resistance was approximately half of the membrane and the share of irreversible/reversible fouling was 88% and 12%. However, the part of reversible fouling becomes predominant when approaching the limiting flux. Permeate fluxes which were obtained allow to estimate that a VRR of approximately 10 could be obtained when concentrating EPS solutions using PES membranes in flat or tubular modules but not in spiral-wound. GRAPHICAL ABSTRACT

Valorization of the Macroalgae Sargassum Muticum by Enzymatic Hydrolysis, Interest of Surfactants to Improve the Extraction of Phlorotannins and Polysaccharides

The use of surfactants to improve enzymatic hydrolysis of the macroalgae Sargassum muticum has been investigated. Visible absorption spectroscopy has been used to quantify the solubilization of both polysaccharides and phlorotannins in the hydrolysates.   After total extraction, results showed that Sargassum muticum contained 2.74% (expressed in percent of the dry weight of the algae) of phlorotannins whose 32 % were in the cell wall. This result shows that it is important to access to the parietal phlorotannins. To reach this objective, we chose the enzymatic approach for destructurating the cell wall of the algae. The use of 5% dry weight (DW - 5% by weight of hydrolyzed algae) of an enzymatic mix containing a commercial beta-glucanase, a commercial protease and an alginate lyase extracted from Pseudomonas alginovora led after 3 hours of hydrolysis to the solubilization of 2.43% DW polysaccharides and 0.52% DW phlorotannins. The use of 0.5% volume of the surfactant Triton® X-100 with 10% DW of the enzymatic mix has allowed to reaching the value of 2.63% DW of solubilized phlorotannins, that is 96% of the total phenolic content.   The use of non-ionic surfactant, combined to enzymatic hydrolysis, showed an increased efficiency in disrupting cell wall and solubilizing phlorotannins in Sargassum muticum.