Identification of Bioactive Peptides from Nannochloropsis oculata Using a Combination of Enzymatic Treatment, in Silico Analysis and Chemical Synthesis.

In vitro ACE-1 inhibitory peptides were characterised previously from a number of microalgal species including Spirulina platensis (peptide IAPG), Chlorella vulgaris (peptides FDL, AFL, VVPPA), Isochrysis galbana (peptide YMGLDLK), Chlorella sorokiniana (peptides IW and LW) and indeed Nannochloropsis oculata (peptides GMNNLTP and LEQ). The isolation of protein from Nannochloropsis oculata using a combination of ammonium salt precipitation and xylanase treatment of resulting biomass combined with molecular weight cut off filtration to produce a permeate and characterisation of bioactive peptides is described. The Angiotensin-1-converting enzyme (ACE-1) IC50 value for the generated permeate fraction was 370 µg/mL. Ninety-five peptide sequences within the permeate fraction were determined using mass spectrometry and eight peptides were selected for chemical synthesis based on in silico analysis. Synthesized peptides were novel based on a search of the literature and relevant databases. In silico, simulated gastrointestinal digestion identified further peptides with bioactivities including ACE-1 inhibitory peptides and peptides with antithrombotic and calcium/calmodulin-dependent kinase II (CAMKII) inhibition. This work highlights the potential of Nannochloropsis oculata biomass as both a protein and bioactive peptide resource, which could be harnessed for use in the development of functional foods and feeds.

Continuous electrocoagulation of Chlorella vulgaris in a novel channel-flow reactor: A pilot-scale harvesting study.

The most frequently used method to harvest microalgae on an industrial scale is centrifugation, although this has very high energy costs. To reduce these costs, a continuous electrocoagulation process for harvesting Chlorella vulgaris was developed and tested using a pilot-scale 111 L working volume device consisting of an electrolyser with iron electrodes, aggregation channel and lamellar settler. The flow rate of the microalgal suspension through the device was 240 L/h. When using controlled cultivation and subsequent electrocoagulation, a high harvesting efficiency (above 85%), a low Fe contamination in the harvested biomass (<4 mg Fe/g dry biomass, a harvested biomass complied with legislative requirements for food) and significant energy savings were achieved. When comparing electrocoagulation and subsequent centrifugation with the use of centrifugation alone, energy savings were 80 % for a biomass harvesting concentration of 0.23 g/L. Electrocoagulation was thus proven to be a feasible pre-concentration method for harvesting microalgae.

Electrocoagulation reduces harvesting costs for microalgae.

Centrifugation is the most commonly used method for harvesting autotrophically produced microalgae, but it is expensive due to high energy demands. With the aim of reducing these costs, we tested electrocoagulation with iron electrodes for harvesting Chlorella vulgaris. During extensive lab-scale experiments, the following factors were studied to achieve a high harvesting efficiency and a low iron content in the harvested biomass: electric charge, initial biomass concentration, pH, temperature, agitation intensity, residual salt content and electrolysis time. A harvesting efficiency greater than 95% was achieved over a broad range of conditions and the residual iron content in the biomass complied with legislative requirements for food. Using electrocoagulation as the pre-concentration step prior to centrifugation, total energy costs were reduced to 0.136 kWh/kg of dry biomass, which is less than 14% of that for centrifugation alone. Our data show that electrocoagulation is a suitable and cost-effective method for harvesting microalgae.

Estimation of Hg(II) in Soil Samples by Bioluminescent Bacterial Bioreporter E. coli ARL1, and the Effect of Humic Acids and Metal Ions on the Biosensor Performance.

Mercury is a ubiquitous environmental pollutant of dominantly anthropogenic origin. A critical concern for human health is the introduction of mercury to the food chain; therefore, monitoring of mercury levels in agricultural soil is essential. Unfortunately, the total mercury content is not sufficiently informative as mercury can be present in different forms with variable bioavailability. Since 1990, the use of bioreporters has been investigated for assessment of the bioavailability of pollutants; however, real contaminated soils have rarely been used in these studies. In this work, a bioassay with whole-cell bacterial bioreporter Escherichia coli ARL1 was used for estimation of bioavailable concentration of mercury in 11 soil samples. The bioreporter emits bioluminescence in the presence of Hg(II). Four different pretreatments of soil samples prior to the bioassay were tested. Among them, laccase mediated extraction was found to be the most suitable over water extraction, alkaline extraction, and direct use of water-soil suspensions. Nevertheless, effect of the matrix on bioreporter signal was found to be severe and not possible to be completely eliminated by the method of standard addition. In order to elucidate the matrix role, influences of humic acid and selected metal ions present in soil on the bioreporter signal were tested separately in laboratory solutions. Humic acids were found to have a positive effect on the bioreporter growth, but a negative effect on the measured bioluminescence, likely due to shading and Hg binding resulting in decreased bioavailability. Each of the tested metal ions solutions affected the bioluminescence signal differently; cobalt (II) positively, iron (III) negatively, and the effects of iron (II) and nickel (II) were dependent on their concentrations. In conclusion, the information on bioavailable mercury estimated by bioreporter E. coli ARL1 is valuable, but the results must be interpreted with caution. The route to functional bioavailability bioassay remains long.