The challenge of water reclamation using membranes in this study was the quite unique wastewater composition resulting from a high share of biotech wastewater. The high content of organic matter and high concentrations of calcium, bicarbonate, and sulphate were considered as challenging for membrane processes. Consequently, an innovative ultra-tight ultrafiltration (u-t UF) membrane was developed and tested on-site at pilot scale. In comparison, a conventional UF and an open nanofiltration (NF) were piloted. The aim was to find the best pre-treatment option for reverse osmosis (RO) to reduce fouling and scaling and produce fit-for-purpose water; for example, cooling. Overall, the quality of the currently used water source was surpassed by the pilot plant. Only a standard post-treatment of the RO permeate was necessary for stabilisation. Results indicated that denser membranes only minimally reduced fouling of RO. An assessment comparing the treatment trains in a life cycle assessment using the data collected from the pilot operation (UF/NF operating settings, RO plant performance, and the design of multi-stage industrial scale RO) revealed lower greenhouse gas emissions compared to seawater desalination. However, if the RO brine treatment becomes mandatory, the greenhouse gas emissions from water reclamation and supply will be higher than those from freshwater supply.