In the present study, we present the aerosol optical properties and radiative forcing (RF) of the tropospheric and stratospheric smoke layers, observed by the Cloud-Aerosol Lidar and Infrared Pathfinder Satellite Observation (CALIPSO) satellite, during the extraordinary Australian biomass burning (BB) event in 2019–2020. These BB layers were studied and analyzed within the longitude range 140° E–20° W and the latitude band 20°–60° S, as they were gradually transported from the Australian banks to the South American continent. These layers were found to be trapped within the Andes circulation, staying for longer time periods in the same longitude region. The BB aerosols reached altitudes even up to 22 km amsl., and regarding their optical properties, they were found to be nearly spherical (particle linear depolarization ratio (PLDR) < 0.10) in the troposphere; while, in the stratosphere, they were more depolarizing with PLDR values reaching up to 0.20. Fine and ultrafine smoke particles were dominant in the stratosphere, according to the observed Ångström exponent, related to the backscatter coefficients obtained by the pair of wavelengths 532 and 1064 nm (Åb up to 3), in contrast to the Åb values in the troposphere (Åb < 1) indicative of the presence of coarser particles. As the aerosols fend off the source, towards North America, a slightly descending trend was observed in the tropospheric Åb values, while the stratospheric ones were lightly increased. A maximum aerosol optical depth (AOD) value of 0.54 was recorded in the lower troposphere over the fire spots, while, in the stratosphere, AOD values up to 0.29 were observed. Sharp changes of carbon monoxide (CO) and ozone (O3) concentrations were also recorded by the Copernicus Atmosphere Monitoring Service (CAMS) in various atmospheric heights over the study region, associated with fire smoke emissions. The tropospheric smoke layers were found to have a negative mean radiative effect, ranging from −12.83 W/m2 at the top of the atmosphere (TOA), to −32.22 W/m2 on the surface (SRF), while the radiative effect of the stratospheric smoke was estimated between −7.36 at the TOA to −18.51 W/m2 at the SRF.