A systematic investigation on gelation self-assembly has been performed on a hydrophobically modulated dipeptide based ambidextrous gelator, N-n-acyl-(L)Val-X(OBn), (X = 1,ω-amino acid). To elucidate the effect of hydrophobic tuning on gelator architecture towards its gelation self-assembly, three sets of gelators with a common formula: C m H 2m+1 C(=O)NH(L)Val(C=O)NH-(CH 2 ) n -(C=O)OBn, were synthesized , Set-I includes gelators with n = 2, m = 9,11,13,15,17, for Set-II it is n = 2,3,5, m=13 and Set-III comprises of two isomeric gelators (n=2, m=15; n=10, m=7). Gelation has been critically analyzed in various apolar (aromatic and aliphatic) and polar (protic and aprotic) solvents using FESEM, CD, IR, WAXRD and rheological studies. Obtained results reveal that π-π type interaction dictates the primary molecular alignment and positioning of amide functionality across the aliphatic chain which influences the peptidic orientation in parallel (when m > n) or antiparallel (when m < n) β-sheet type organization in their self-assembly. The thermal stability, gelation number (GN), T gel and yield stress of gel systems increases with m, but for a given m, the trend goes apparently inverse with increasing n. Circular dichroism (CD) studies suggest an intriguing evidence of non-planarity of amide plane during selfassembly, highlighting the involvement of conformational change taking place during molecular organization towards its gelation. Despite complex nature of solvent-gelator interaction, the effect of Hbonding component of solubility parameters was found to have a significant role on self-assembly.Overall, supramolecular forces acting at specific functionalities encrypted in gelator backbone must overcome the solvation energy with synergic assistance of solvophobic effect towards stabilization of gel-network with optimum gelator backbone conformation for achieving required enthalpic contribution for self-assembly.