Systems-wide analysis of acclimation responses to long-term heat stress and recovery in the photosynthetic model organism Chlamydomonas reinhardtii.

We applied a top-down systems biology approach to understand how Chlamydomonas reinhardtii acclimates to long-term heat stress (HS) and recovers from it. For this, we shifted cells from 25 to 42°C for 24 h and back to 25°C for ≥8 h and monitored abundances of 1856 proteins/protein groups, 99 polar and 185 lipophilic metabolites, and cytological and photosynthesis parameters. Our data indicate that acclimation of Chlamydomonas to long-term HS consists of a temporally ordered, orchestrated implementation of response elements at various system levels. These comprise (1) cell cycle arrest; (2) catabolism of larger molecules to generate compounds with roles in stress protection; (3) accumulation of molecular chaperones to restore protein homeostasis together with compatible solutes; (4) redirection of photosynthetic energy and reducing power from the Calvin cycle to the de novo synthesis of saturated fatty acids to replace polyunsaturated ones in membrane lipids, which are deposited in lipid bodies; and (5) when sinks for photosynthetic energy and reducing power are depleted, resumption of Calvin cycle activity associated with increased photorespiration, accumulation of reactive oxygen species scavengers, and throttling of linear electron flow by antenna uncoupling. During recovery from HS, cells appear to focus on processes allowing rapid resumption of growth rather than restoring pre-HS conditions.

Systems Analysis of the Response of Photosynthesis, Metabolism, and Growth to an Increase in Irradiance in the Photosynthetic Model Organism Chlamydomonas reinhardtii.

We investigated the systems response of metabolism and growth after an increase in irradiance in the nonsaturating range in the algal model Chlamydomonas reinhardtii. In a three-step process, photosynthesis and the levels of metabolites increased immediately, growth increased after 10 to 15 min, and transcript and protein abundance responded by 40 and 120 to 240 min, respectively. In the first phase, starch and metabolites provided a transient buffer for carbon until growth increased. This uncouples photosynthesis from growth in a fluctuating light environment. In the first and second phases, rising metabolite levels and increased polysome loading drove an increase in fluxes. Most Calvin-Benson cycle (CBC) enzymes were substrate-limited in vivo, and strikingly, many were present at higher concentrations than their substrates, explaining how rising metabolite levels stimulate CBC flux. Rubisco, fructose-1,6-biosphosphatase, and seduheptulose-1,7-bisphosphatase were close to substrate saturation in vivo, and flux was increased by posttranslational activation. In the third phase, changes in abundance of particular proteins, including increases in plastidial ATP synthase and some CBC enzymes, relieved potential bottlenecks and readjusted protein allocation between different processes. Despite reasonable overall agreement between changes in transcript and protein abundance (R2 = 0.24), many proteins, including those in photosynthesis, changed independently of transcript abundance.

Evidence for a role of VIPP1 in the structural organization of the photosynthetic apparatus in Chlamydomonas.

The vesicle-inducing protein in plastids (VIPP1) was suggested to play a role in thylakoid membrane formation via membrane vesicles. As this functional assignment is under debate, we investigated the function of VIPP1 in Chlamydomonas reinhardtii. Using immunofluorescence, we localized VIPP1 to distinct spots within the chloroplast. In VIPP1-RNA interference/artificial microRNA cells, we consistently observed aberrant, prolamellar body-like structures at the origin of multiple thylakoid membrane layers, which appear to coincide with the immunofluorescent VIPP1 spots and suggest a defect in thylakoid membrane biogenesis. Accordingly, using quantitative shotgun proteomics, we found that unstressed vipp1 mutant cells accumulate 14 to 20% less photosystems, cytochrome b(6)f complex, and ATP synthase but 30% more light-harvesting complex II than control cells, while complex assembly, thylakoid membrane ultrastructure, and bulk lipid composition appeared unaltered. Photosystems in vipp1 mutants are sensitive to high light, which coincides with a lowered midpoint potential of the Q(A)/Q(A)(-) redox couple and increased thermosensitivity of photosystem II (PSII), suggesting structural defects in PSII. Moreover, swollen thylakoids, despite reduced membrane energization, in vipp1 mutants grown on ammonium suggest defects in the supermolecular organization of thylakoid membrane complexes. Overall, our data suggest a role of VIPP1 in the biogenesis/assembly of thylakoid membrane core complexes, most likely by supplying structural lipids.

Heterotrophic growth of Galdieria sulphuraria on residues from aquaculture and fish processing industries.

The study aimed at zero-waste utilization of fish processing streams for cultivation of microalgae Galdieria sulphuraria. Wastewater from a fish processing facility, slam (mix of used fish feed and faeces), and dried pellet (sediments after enzymatic hydrolysis of rainbow trout) were investigated as potential sources of carbon, nitrogen, and phosphate for cultivation of G. sulphuraria. The pellet extract was found to support the growth of G. sulphuraria when appropriate diluted, at concentrations below 40 % (v/v). It was revealed that wastewater does not impact the growth negatively, however free amino nitrogen and carbon sources need to be supplied from another source. Therefore, only proteolyzed pellet extract (20 %, v/v) was selected for upscaling and a biomass concentration of 80 g L-1 (growth rate was 0.72 day-1) was achieved in a non-sterile fed-batch culture. Even though biomass was produced under non-sterile conditions no pathogens such as Salmonella sp. could be detected.

Transformation of Seafood Side-Streams and Residuals into Valuable Products.

Seafood processing creates enormous amounts of side-streams. This review deals with the use of seafood side-streams for transformation into valuable products and identifies suitable approaches for making use of it for different purposes. Starting at the stage of catching fish to its selling point, many of the fish parts, such as head, skin, tail, fillet cut-offs, and the viscera, are wasted. These parts are rich in proteins, enzymes, healthy fatty acids such as monounsaturated and polyunsaturated ones, gelatin, and collagen. The valuable biochemical composition makes it worth discussing paths through which seafood side-streams can be turned into valuable products. Drawbacks, as well as challenges of different aquacultures, demonstrate the importance of using the various side-streams to produce valuable compounds to improve economic performance efficiency and sustainability of aquaculture. In this review, conventional and novel utilization approaches, as well as a combination of both, have been identified, which will lead to the development of sustainable production chains and the emergence of new bio-based products in the future.