Spectra manager program

The circular dichroism spectrum of rBASIDIN was collected from 190 to 240 ηm using a J-815 spectropolarimeter (Jasco), with a thermostatically controlled stand with a PTC-423S/15 Peltier device. For this, we used 300μLof the protein at a molar concentration of 0.05 μmol L⁻¹; in Tris-HCl buffer 5 mmol L⁻¹, pH 7.4. The scans were made with the Spectra Manager program (Jasco), at a speed of 50 ηm min⁻¹ and a collection interval of 1.0 ηm. Readings were taken at temperatures of 25 and 95 °C, using a polarized quartz cuvette with a 1 mm path length. The spectra obtained were the results of six consecutive scans, and the data from the spectra were analyzed with the K2D3 program to estimate the percentage of the secondary structure of the recombinant protein rBASIDIN.

CD analysis was performed on a Jasco 810 spectropolarimeter (Jasco Analytical Instrument, Easton, MD) under nitrogen. CD spectra were measured at room temperature (RT, 25°C) in a 1 mm path length quartz cell. Spectra of VlsE (10 μM) or VlsE_ECM (10 μM) were recorded in phosphate based saline buffer (PBS) at RT, and three far-UV CD spectra were recorded from 190 to 250 nm for far-UV CD in 1 nm increments. The background spectrum of PBS without proteins was subtracted from the protein spectra. CD spectra were initially analyzed by the software Spectra Manager Program (Jasco). Analysis of spectra to extrapolate secondary structures were performed using the K2D3 analysis programs [58 (link)].

His-AuNCs were prepared in a microwave reactor (Monowave 300, Anton Paar, Graz, Austria). The absorption spectra were obtained with a JASCO V-670 UV-Vis-NIR spectrophotometer (Tokyo, Japan) in a quartz cuvette of 2 mm from Helma and analyzed with the Spectra Manager program (JASCO). HR-TEM images were acquired with a Tecnai TEM microscope (Tecnai G2 F20 X-TWIN, FEI Company, Hillsboro, OR, USA) at 200 kV accelerating voltage. The zeta potential of His-AuNCs was measured using the Nano ZS-900 Zetasizer from Malvern. The fluorescence spectra were measured with a JASCO FP6500 spectrofluorometer (Tokyo, Japan) having 1 nm spectral resolution and using a 150 W Xenon lamp as excitation source. The fluorescence spectra of His-AuNCs were obtained with excitation and emission bandwidths fixed at 3 nm. Helma quartz cuvettes (5 × 5 mm) were used for all fluorescence measurements, while the obtained spectra were analyzed with the Spectra Manager software. All photos regarding the paper-based detection assays were captured in a dark room with a smartphone camera with the following settings: ISO at 100, speed at A 1/10, exposure value at −0.8 and white balance at A 4400 K.