Höfundur: Kristín Edda Gylfadóttir

The identification of reliable biomarkers, such as amino acids, is key for the search of extraterrestrial life. A large number of microorganisms metabolize, synthesize, take up and excrete amino acids as part of the amino acid metabolism during aerobic and/or anaerobic respiration or in fermentation. In this work, we investigated whether the anaerobic microbial metabolism of amino acids could leave a secondary biosignature indicating biological activity in the environment around the cells. The observed fingerprints would reflect the physiological capabilities of the specific microbial community under investigation. The metabolic processing of an amino acid mixture by two distinct anaerobic microbial communities collected from Islinger Mühlbach (ISM) and Sippenauer Moor (SM), Germany was examined. The amino acid mixture contained L-alanine, β-alanine, L-aspartic acid, DL-proline, L-leucine, L-valine, glycine, L-phenylalanine and L-isoleucine. In parallel, an amino acid spiked medium without microorganisms was used as a control to determine abiotic changes over time. Liquid chromatography mass spectrometry (LC-MS) was used to track amino acid changes over time. When comparing to the control samples that did not show significant changes of amino acids concentrations over time, we found that glycine was almost completely depleted from both microbial samples to less than 3% after the first two weeks- This results indicates a preferential use of this simple amino acid by these microbial communities. Although glycine degradation can be caused by abiotic processes, these results show that its preferential depletion in an environment would be consistent with the presence of life. We found changes in most other amino acids that varied between amino acids and communities, suggesting complex dynamics with no clear universal pattern that might be used as a signature of life. However, marked increases in amino acids, caused by cellular synthesis and release into the extracellular environment (e.g., alanine), were observed and could be considered a signature of metabolic activity. We conclude, that substantial anomalous enhancements of some amino acids against the expected abiotic background concentration may be an agnostic signature of the presence of biological processes.

Seaweed (or macroalgae) produced sustainably at large scale opens opportunities as source of fuels, chemicals and food. The production does not directly compete with terrestrial food production and may make use of anthropogenic sources of carbon dioxide and nitrogen. Seaweed biomass can be transformed into a suitable substrate for fermentation using a biorefinery approach. In this study the entire process of biofuel production from seaweed is described: starting with cultivation and harvest, the seaweed is dried and cut, enzymatically hydrolysed, demineralized, detoxified, and finally fermented into acetone, butanol, and ethanol (ABE). Juvenile Saccharina latissimawas directly seeded on AlgaeTex® nets and cultivated in the North East Atlantic off the west coast of Scotland for 6 months. Sun dried seaweed was hydrolysed with different enzymes, looking for optimal glucose release, solid/liquid ratio, and enzyme load. Using Cellic® CTec2 in combination with alginate lyases, approximately 80% of available glucose was released. The hydrolysis was scaled up to 100 L, using only Cellic® CTec2. Part of the hydrolysate was demineralized using ion-exclusion chromatography, removing over 90% of minerals while recovering 92% of glucose and mannitol. A fraction of the demineralized hydrolysate was additionally detoxified using a hydrophobic resin to remove hydrophobic components to a concentration below detection limit. The three hydrolysates (untreated, demineralized, and demineralized followed by detoxification) were used as substrate for ABE production by a newly developed strain of Clostridium acetobutylicum adapted to grow on S. latissima hydrolysate. Demineralization reduced the lag phase of fermentation from 72 h (untreated) to 24–48 h. Further detoxification of the hydrolysate led to immediate fermentation, resulting in a yield of 0.23 ± 0.02 g_ABE/g_sugarsimilar to control fermentation in control medium (0.19 g_ABE/g_sugar).

In addition to their use in human medicine, antimicrobials are also used in food animals and aquaculture, and their use can be categorized as therapeutic against bacterial infections. The use of antimicrobials in aquaculture may involve a broad environmental application that affects a wide variety of bacteria, promoting the spread of bacterial resistance genes. Probiotics and bacteriocins, antimicrobial peptides produced by some types of lactic acid bacteria (LAB), have been successfully tested in aquatic animals as alternatives to control bacterial infections. Supplementation might have beneficial impacts on the intestinal microbiota, immune response, development, and/or weight gain, without the issues associated with antibiotic use. Thus, probiotics and bacteriocins represent feasible alternatives to antibiotics. Here, we provide an update with respect to the relevance of aquaculture in the animal protein production sector, as well as the present and future challenges generated by outbreaks and antimicrobial resistance, while highlighting the potential role of probiotics and bacteriocins to address these challenges. In addition, we conducted data analysis using a simple linear regression model to determine whether a linear relationship exists between probiotic dose added to feed and three variables of interest selected, including specific growth rate, feed conversion ratio, and lysozyme activity.

Enzymatic hydrolysis is a novel method to recover highly potent bioactive fish protein hydrolysates (FPHs) from fish processing side-streams. The common way of producing FPHs directly from fish side-streams may be inappropriate due to the excess of lipids and pro-oxidants, especially in lipid-rich streams, as obtained from Tra catfish. This study aimed to optimise the hydrolysis conditions for a commercial enzyme (Alcalase® 2.4 L) (enzyme concentrate, temperature, and time) in FPH production from the fish protein isolate obtained from Tra catfish dark muscle (DM-FPI) using the pH-shift method. The degree of hydrolysis (DH), protein recovery (PR), and antioxidant properties, including DPPH radical scavenging activity (DPPH-RSA) and total reducing power capacity (TRPC), were measured to evaluate the effects of the hydrolysis conditions on the FPHs. Optimal hydrolysis was obtained at an enzyme/substrate protein ratio of 3% (v/w) and a hydrolysis temperature of 50 °C for 3 h. The FPHs obtained from different substrates, including DM-FPI, abdominal cut-off (ACO) FPI, and head and backbone blend (HBB) FPI, had similar DHs under these optimum conditions, ranging from 22.5% to 24.0%. However, the FPH obtained from abdominal cut-off isolate (ACO-FPH) showed the highest PR of 81.5 ± 4.3% and the highest antioxidant properties, with a DPPH-RSA of 86.1 ± 1.6% and a TRPC of 6.4 ± 0.4 equivalent mg vitamin C/g protein. The resulting FPHs present a natural source of antioxidants with great potential for food applications, especially the ACO-FPH. In addition, all FPHs had excellent amino acid profiles, indicating strong potential for their use as supplements. Tra catfish protein-rich side-streams can thus be processed into high-value bioactive FPHs using Alcalase for human consumption.

Quality changes of protein and non-protein nitrogen compounds during industrial fishmeal processing of fatty pelagic species (mackerel/herring rest material blend, MHB) and lean fish (whole blue whiting, BW) were studied to identify processing steps that require optimization to allow production of products for human consumption. Samples from protein-rich processing streams throughout the fishmeal production were analyzed for proximate composition, salt soluble protein content (SSP), biogenic amines (BA), total volatile basic nitrogen (TVB-N), trimethylamine (TMA), and dimethylamine (DMA). Mass flows throughout processing were balanced based on the total mass and proximate composition data. The quality of the final fishmeal products was highly dependent on the fish species being processed, indicating that the processes require optimization towards each raw material. The chemical composition changed in each processing step, resulting in different properties in each stream. Most of the non-protein nitrogen compounds (including BA, TVB-N, TMA, and DMA) followed the liquid streams. However, the concentrate contributed less than 20% to the produced fishmeal quantity. Mixing of this stream into the fishmeal processing again, as currently carried out, should thus be avoided. Furthermore, the cooking, separating, and drying steps should be optimized to improve the water and lipid separation and avoid the formation of undesired nitrogen compounds to produce higher-value products intended for human consumption.

This chapter provides a summary of food production in three regions of the Arctic. These include: the entirety of Iceland; Norway’s three northernmost counties—Nordland, Troms, and Finnmark; and northern Canada, including Yukon, Northwest Territories, Nunavut, Nunavik, and Labrador. In 2016, the Sustainable Development Working Group endorsed The Arctic as a Food Producing Region research project. The export of fish and fish products are by far the most important export items from Iceland and contribute significantly to the Icelandic economy. From 2012 to 2016, Norway exported fish products worth 339,207,335,000 Norwegian krone. Marine products accounted for 89% of northern Canada’s total food export. There are over 3,000 sheep farms in Iceland. These farms tend to be small and family owned. The sheep farming is as old as the human settlement of Iceland. The largest agricultural production systems in Northern Norway are based on meat production. The agricultural production in northern Norway is the northernmost active agricultural system in the world.

Background

The marine thermophilic bacterium Rhodothermus marinus can degrade many polysaccharides which makes it interesting as a future cell factory. Progress using this bacterium has, however, been hampered by limited knowledge on media and conditions for biomass production, often resulting in low cell yields and low productivity, highlighting the need to develop conditions that allow studies of the microbe on molecular level. This study presents development of defined conditions that support growth, combined with evaluation of production of carotenoids and exopolysaccharides (EPSs) by R. marinus strain DSM 16675.

Results

Two defined media were initially prepared: one including a low addition of yeast extract (modified Wolfe’s medium) and one based on specific components (defined medium base, DMB) to which two amino acids (N and Q), were added. Cultivation trials of R. marinus DSM 16675 in shake flasks, resulted in maximum cell densities (OD_620 nm) of 2.36 ± 0.057, cell dry weight (CDW) 1.2 ± 0.14 mg/L, total carotenoids 0.59 × 10^–3 mg/L, and EPSs 1.72 ± 0.03 mg/L using 2 g/L glucose in DMB. In Wolfe’s medium (supplemented by 0.05 g/L yeast extract and 2.5 g/L glucose), maximum OD_620 nm was 2.07 ± 0.05, CDW 1.05 ± 0.07 mg/L, total carotenoids 0.39 × 10^–3 mg/L, and EPSs 1.74 ± 0.2 mg/L. Growth trials at 5 g/L glucose in these media either failed or resulted in incomplete substrate utilization. To improve reproducibility and increase substrate utilization, a screening of macroelements (e.g. phosphate) in DMB, was combined with use of trace elements and vitamins of the modified Wolfe’s medium. The resulting defined minimal R. marinus medium, (DRM), allowed reproducible cultivations to a final OD_620nm of 6.6 ± 0.05, CDW 2.85 ± 0.07 mg/L, a maximum specific growth rate (µ_max) of 0.26 h⁻¹, total carotenoids 0.77 × 10^–3 mg/L and EPSs 3.4 ± 0.17 mg/L in cultivations supplemented with up to 5 g/L glucose.

Conclusion

A minimal defined medium (DRM) was designed that resulted in reproducible growth and an almost doubled formation of both total carotenoids and EPSs. Such defined conditions, are necessary for systematic studies of metabolic pathways, to determine the specific requirements for growth and fully characterize metabolite production.