N,N,N-trimethyl chitosan (TMC), quaternized hydrophilic derivative of chitosan, has been projected to have wide applications in the pharmaceutical industry owing to its improved solubility at physiological conditions. However, the conventional synthesis of TMC involves toxic organic agents, which complicates its use for biological applications. Moreover, these reactions result into unwanted O-methylation and scission of the parent polymer. In the present study we have addressed these limitations by employing a green approach to synthesize TMC, by using lipase as the biocatalyst and dimethyl carbonate (DMC) as the green methylating agent, in a reaction medium comprising of ternary deep eutectic solvents (TDESs). Synthesis of TMC was carried out by using two different lipases from Burkholderia cepacia and Candida rugosa. The resulting TMC was characterized by using FTIR, H NMR, DSC, XRD. Methylation was confirmed by FTIR analysis (-CH at 1666 cm) and H NMR (?? = 3.3 ppm). DSC study revealed a lower thermal stability of TMC as compared to chitosan. These results indicated the possibility of using DMC as a green methylating agent, along with TDESs as green and sustainable solvents, for lipase catalyzed reactions. TMC was successfully synthesized and exhibited a degree of quaternization of about 12.5%, 15.69%, when synthesized used lipases from Burkholderia cepacia and Candida rugosa, respectively.