Today, the development of green nanocatalysts is among the popular topics due to the need for energy production and the cleaning of organic pollutants. In this approach, Bacillus thuringiensis, a bacterium, was used as a biosupport of ruthenium/nickel co-doped zinc nanoparticles (btRNZn NPs) to release hydrogen from the methanolysis of sodium borohydride (NaBH 4 ). In addition, their photocatalytic activity was reported against Methyl Orange (MO) organic dye. This study focused on the preparation, characterization, and catalytic and photocatalytic activity of the btRNZn biocatalyst for the release of hydrogen from the methanolysis of NaBH 4 and removal of MO dye. According to TEM analysis, the average size of btRNZn NPs was found to be 11.78 nm; in addition, btRNZn NPs showed a photodegradation effect of 68.2% against MO dye at 90 min, and its photocatalytic mechanism was discussed. The effects of the catalyst, substrate, and temperature in the methanolysis reaction of NaBH 4 in the presence of the catalyst were investigated extensively. The reaction kinetics was calculated, and TOF, activation energy, and enthalpy energy were measured as 2497.14 h −1 , 14.89 kJ/mol, and 12.35 kJ/mol, respectively. It was observed that the methanolysis process is a first-order reaction based on the amount of the catalyst and substrate. This study aimed to synthesize a nanobiocatalyst (btRNZn NPs) by a biological method, and it will be used as a great photocatalyst to prevent wastewater pollution; also, it can be an excellent catalyst to produce hydrogen from NaBH 4 methanolysis. The application of btRNZn NPs in solar photocatalysis to prevent wastewater pollution and to research it for energy production through hydrogen creation are both made clear by these studies.