Fluorescent Labeling of Outer Membrane Proteins Using the SpyCatcher-SpyTag System.

The SpyCatcher-SpyTag system has become a popular and versatile tool for protein ligation. It is based on a small globular protein (SpyCatcher) that binds to a 13-residue peptide (SpyTag), which subsequently leads to the formation of a covalent isopeptide bond. Thus, the reaction is essentially irreversible. Here, we describe how the SpyCatcher-SpyTag system can be used to label surface-exposed bacterial outer membrane proteins, e.g., for topology mapping or fluorescent time-course experiments. We cover using fluorescence measurements and microscopy to measure labeling efficiency using SpyCatcher fused with superfolder GFP in this chapter.

A sediment diagenesis model on sediment oxygen demand in managing eutrophication on Taihu, China.

Blue-green algae (CyanoHABs), photosynthetic bacteria that create a harmful aquatic environment, have been a trending issue on Taihu for over a decade. CyanoHABs adapt to varying climatic changes, which explains why the problem on Taihu still thrives. One major drive that keeps the algae is Sediment Oxygen Demand (SOD). In this paper, seasonal and spatial variations of SOD that contribute immensely to nutrient growth in Lake Taihu were done using the Environmental Fluid Dynamics Code (EFDC). The results were analyzed based on Nitrogenous SOD (NSOD) and Total SOD (TSOD). Summer results ranged from - 0.05754 to - 0.0826 (- 0.75658 to - 0.83902) (g/m2/day) and Winter values ranged from - 0.3022 to - 0.40171 (- 1.34486 to - 1.48856) (g/m2/day) indicate a gradual decrease in NSOD (TSOD) values respectively. Relatively higher values in summer are attributed to warmer surface water which sets up thermal stratification to increase the internal loading of nitrogen. Lower winter values are related to inverse stratification, where lower oxygen concentration decreases the SOD to trigger ammonium accumulation in the water column. NSOD (TSOD) values for Autumn results ranged from - 0.1039 to - 0.24786 (- 0.96251 to - 1.39454) (g/m2/day) and Spring values of - 0.43019 to - 0.35959 (- 1.48297 to - 0.54089) (g/m2/day). Transition seasons (i.e., Autumn and Spring) results are impacted by wind mixing that allows dissolved oxygen and nutrients in the whole water column. However, spring values depict a gradual increase in SOD value attributed to spring turnover and gradual stratification, which decrease nutrient concentration. In contrast, decreasing SOD values in autumn are related to mixing, but temperature decreases tend to increase nutrient concentrations. Carbonaceous sediment oxygen demand (CSOD), due to sulfide oxidation, presents high values from the difference between TSOD and NSOD. Based on the high values of CSOD, it is highly recommended that more research on eutrophic Taihu lakes would consider delving into CSOD.