AbstractTemperature is one of the most important parameters in biohydrogen production by way of photo-fermentation. Enzymatic hydrolysate of corncob powder was utilized as a substrate. Computational fluid dynamics (CFD) modeling was conducted to simulate the temperature distribution in an up-flow baffle photo-bioreactor (UBPB). Commercial software, GAMBIT, was utilized to mesh the photobioreactor geometry, while the software FLUENT was adopted to simulate the heat transfer in the photo-fermentation process. The inlet velocity had a marked impact on heat transfer; the most optimum velocity value was 0.0036 m•s-1 because it had the smallest temperature fluctuation and the most uniform temperature distribution. When the velocity decreased from 0.0036 m•s-1 to 0.0009 m•s-1, more heat was accumulated. The results obtained from the established model were consistent to the actual situation by comparing the simulation values and experimental values. The hydrogen production simulation verified that the novel UBPB was suitable for biohydrogen production by photosynthetic bacteria because of its uniform temperature and lighting distribution, with the serpentine flow pattern also providing mixing without additional energy input, thus enhancing the mass transfer and biohydrogen yield.