An unexpected role for leucyl aminopeptidase in UV tolerance revealed by a genome-wide fitness assessment in a model cyanobacterium.

UV radiation (UVR) has significant physiological effects on organisms living at or near the Earth's surface, yet the full suite of genes required for fitness of a photosynthetic organism in a UVR-rich environment remains unknown. This study reports a genome-wide fitness assessment of the genes that affect UVR tolerance under environmentally relevant UVR dosages in the model cyanobacterium Synechococcus elongatus PCC 7942. Our results highlight the importance of specific genes that encode proteins involved in DNA repair, glutathione synthesis, and the assembly and maintenance of photosystem II, as well as genes that encode hypothetical proteins and others without an obvious connection to canonical methods of UVR tolerance. Disruption of a gene that encodes a leucyl aminopeptidase (LAP) conferred the greatest UVR-specific decrease in fitness. Enzymatic assays demonstrated a strong pH-dependent affinity of the LAP for the dipeptide cysteinyl-glycine, suggesting an involvement in glutathione catabolism as a function of night-time cytosolic pH level. A low differential expression of the LAP gene under acute UVR exposure suggests that its relative importance would be overlooked in transcript-dependent screens. Subsequent experiments revealed a similar UVR-sensitivity phenotype in LAP knockouts of other organisms, indicating conservation of the functional role of LAPs in UVR tolerance.

Controls on the distribution of fluorescent dissolved organic matter during an under-ice algal bloom in the western Arctic Ocean.

In this study we used fluorescence excitation and emission matrix spectroscopy, hydrographic data, and a self-organizing map (SOM) analysis to assess the spatial distribution of labile and refractory fluorescent dissolved organic matter (FDOM) for the Chukchi and Beaufort Seas at the time of a massive under-ice phytoplankton bloom during early summer 2011. Biogeochemical properties were assessed through decomposition of water property classes and sample classification that employed a SOM neural network-based analysis which classified 10 clusters from 269 samples and 17 variables. The terrestrial, humic-like component FDOM (ArC1, 4.98 ± 1.54 Quinine Sulfate Units (QSU)) and protein-like component FDOM (ArC3, 1.63 ± 0.88 QSU) were found to have elevated fluorescence in the Lower Polar Mixed Layer (LPML) (salinity ~29.56 ± 0.76). In the LPML water mass, the observed contribution of meteoric water fraction was 17%, relative to a 12% contribution from the sea ice melt fraction. The labile ArC3-protein-like component (2.01 ± 1.92 QSU) was also observed to be elevated in the Pacific Winter Waters mass, where the under-ice algal bloom was observed (~40-50 m). We interpreted these relationships to indicate that the accumulation and variable distribution of the protein-like component on the shelf could be influenced directly by sea ice melt, transport, and mixing processes and indirectly by the in situ algal bloom and microbial activity. ArC5, corresponding to what is commonly considered marine humic FDOM, indicated a bimodal distribution with high values in both the freshest and saltiest waters. The association of ArC5 with deep, dense salty water is consistent with this component as refractory humic-like FDOM, whereas our evidence of a terrestrial origin challenges this classic paradigm for this component.

Phenotypically complex living materials containing engineered cyanobacteria.

The field of engineered living materials lies at the intersection of materials science and synthetic biology with the aim of developing materials that can sense and respond to the environment. In this study, we use 3D printing to fabricate a cyanobacterial biocomposite material capable of producing multiple functional outputs in response to an external chemical stimulus and demonstrate the advantages of utilizing additive manufacturing techniques in controlling the shape of the fabricated photosynthetic material. As an initial proof-of-concept, a synthetic riboswitch is used to regulate the expression of a yellow fluorescent protein reporter in Synechococcus elongatus PCC 7942 within a hydrogel matrix. Subsequently, a strain of S. elongatus is engineered to produce an oxidative laccase enzyme; when printed within a hydrogel matrix the responsive biomaterial can decolorize a common textile dye pollutant, indigo carmine, potentially serving as a tool in environmental bioremediation. Finally, cells are engineered for inducible cell death to eliminate their presence once their activity is no longer required, which is an important function for biocontainment and minimizing environmental impact. By integrating genetically engineered stimuli-responsive cyanobacteria in volumetric 3D-printed designs, we demonstrate programmable photosynthetic biocomposite materials capable of producing functional outputs including, but not limited to, bioremediation.