The disclosure relates to a process of obtaining a fully folded two chain insulin glargine that require no further processing to make it functionally active. The present disclosure discloses a surprising effect of over expression of Kex2p intracellularly under the control of inducible FLD1 promoter in the host Pichia pastoris to produce two chain functional glargine secreted directly in the medium. The schematic diagram of how the two chains are made inside the host Pichia pastoris and secretes into the medium.