Expanding the use of tropical biomass wastes for nitrogen-limited fermentation was investigated, specifically, the production of medium chain length polyhydroxyalkanoates. Comprehensive central composite design was conducted to assess pH, temperature, biomass solid loading, cellulase loading and amylase loading and their impact on the hydrolysis of palm, coconut and cassava wastes. Glucose yields of 33.3, 31.7 and 79.0 % wt. with respect to total glucose for palm, coconut and cassava, respectively. Importantly, the impact on the total nitrogen derived during enzymatic hydrolysis of these tropical biomass was described for the first time. The level of nitrogen needs to be properly controlled as high nitrogen would result in low carbon to nitrogen ratio leading to low polyhydroxyalkanoates accumulation, but low nitrogen would hinder growth of the biopolymer producer. Maximum hydrolysate nitrogen, were 1.80, 1.55 and 0.871 g/l for palm, coconut and cassava, respectively. Using the surface responses, biomass media designed for high carbon-to-nitrogen were produced and validated using Pseudomonas putida. Low glucose-carbon to nitrogen were found for palm and coconut after scale-up, leading to the majority of their polyhydroxyalkanoates not being biomass-derived. However, cassava-derived biopolymers were successfully accumulated at 9.01 and 7.13 % wt. for total medium chain length polyhydroxyalkanoates and 10-carbon polyhydroxyalkanoates, respectively. This study provides an important foundation for the expansion of tropical biomass wastes for biopolymer production and other nitrogen-limited applications in general.