Coumarins and their derivatives comprise an important class of secondary metabolites produced by a large range of plant sources. Literature reports different types of biological activities for these compounds and among them are the antimicrobial, anticoagulant, estrogenic, anti-inflammatory, anti-HIV, antioxidant, and other. As promising therapeutic agents, new and sustainable methodologies for those synthesis may gain even more prominence with a green and lower cost approach [1,2]. So, the derivatization of a coumarin skeleton can improve the activity as already observed in literature for other phenolic compound, such as flavonoids. The present report describes the enzymatic acylation of umbelliferone with different vinyl esters as acyl donors biocatalyzed by the commercial lipase Novozym® 435 and by an endophytic fungi immobilized in zeolite. It also aims the investigation for their antibacterial activity against ATCC and clinical strains isolated from hospital infection sites. The umbelliferone esters (1―5) were synthesized through the acylation reaction of 7-hydroxy-2H-chromen-2-one with different long chain vinyl esters catalyzed by the lipase Novozym 435. The reaction conditions were: 10 % Novozym 435; tetrahydrofuran:acetone (3 (solvent):1(cosolvent)) for the reactions with acetate, propionate and butyrate vinyl esters, and (9 (solvent):1(co-solvent)) for decanoate and laurate vinyl esters; acyl donor /umbelliferone molar ratio of 10:1 and 60 °C. Aliquots (20 μL) were withdrawn from 24 to 168 h and analyzed by GC-MS. All tumbelliferone esters were characterized by Nuclear Magnetic Resonance (NMR) and high resolution mass spectrometry (HRMS). The recycle of the reactions will be evaluated for both biocatalysts. The esters with short chain achieved 50―90 % conversion while with the long chain vinyl esters achieved 10 – 15 % conversion. The antibacterial activity of the products were tested in order to determine the minimum inhibitory concentration (MIC) against ATCC strains (S. aureus ATCC 33591 and ATCC 29213; S. epidermidis ATCC 12228; S. hominis ATCC 27844; S. simulans ATCC 27851; S. saprophyticus ATCC 15305; Pseudomonas aeruginosa ATCC 27853; Klebsiella pneumoniae ATCC 700603; E. coli ATCC 25922) and clinical strains (CS) (S. aureus CS 601, CS 664, CS527, CS 551, CS 517 CS 1025; Acinetobacter spp. CS OXA-51; A. baumannii CS 55a). The results displayed by 7-laurate and 7-decanoate-umbelliferone esters showed the highest antibacterial potential, with 1 mM inhibitory activity for ATCC 33591, a methicillin and oxacillin resistant Staphylococcus aureus strain. They were also able to inhibit gram-negative bacterial strains, such as P. aeruginosa (MIC: 0.5 mM) and K. pneumoniae (MIC:1 mM). In addition, 7-laurate- and 7-decanoate-umbelliferone esters were able to inhibit all clinical strains (MIC: 1mM; except 7-laurate-umbelliferone in which MIC: 0.5 mM against 55a). The reactions with the endophytic fungi immobilized in zeolite (rich in lipases) will be also conducted and the conversion rates established. This is the first study performing the biocatalysis of umbelliferone followed by the purification of the products and the antibacterial evaluation.