<p>Transition from conventional packaging to biodegradable edible film mitigate the adverse environmental impacts and promote a more sustainable and circular economy. This study aimed to investigate the effect of different concentrations of microbial transglutaminase (MTGase) enzyme (0.14 and 0.28&#xa0;g/100&#xa0;mL of film-forming solution) on the physiochemical, mechanical properties, surface morphology, and anti-bacterial properties of nano-emulsified kesum essential oil-based gelatine-chitosan edible film. The gelatine-chitosan film with MTGase concentration at 0.28&#xa0;g/100&#xa0;mL of film-forming solution demonstrate desirable thickness (0.25&#xa0;mm), low moisture content (3.58%), low water solubility (44.8%), low water vapour permeability (7.61 × 10^–7&#xa0;g&#xa0;mm/m² Pa h), high elongation at break (631.13%), low Young’s modulus (0.0009&#xa0;MPa), and homogenous structure in cross-sectional micrographs. The addition of MTGase in film had no impact on the anti-bacterial properties. The addition of MTGase at 0.28&#xa0;g/100&#xa0;mL of film-forming solution in gelatine-chitosan film was able to improve the physicochemical properties of them film, producing a flexible and functional edible film. This demonstrates its potential to be a promising active edible packaging for food applications.</p> Graphical abstract <p></p>