EnZymClass: Substrate specificity prediction tool of plant acyl-ACP thioesterases based on ensemble learning

•A machine learning classifier, EnZymClass, was developed to predict substrate specificity of plant acyl-ACP thioesterases.•Two thioesterases with previously uncharacterized medium-chain substrate activity in E. coli were identified using EnZymClass.•The thioesterase ClFatB3 was further engineered to exhibit a 4.2 fold improvement in ten-carbon free fatty acid production.