A single-stranded ribosome inactivating protein SoRIP1 from Spinacia oleracea (Spinach) is a 30 kDa protein. It is a multipurpose protein with versatile biological activities. This paper defines cloning and expression of SoRIP1 in bacteria to study its versatile biological properties and cytotoxic potential. The recombinant plasmid pET29a-SoRIP1 was synthesized and then transferred into top10 and BL21 (DE3) cells of Escherichia coli. The protein SoRIP1 was expressed after giving induction of 0.5 mM IPTG for 16 h at 37 ℃. The purification was achieved by Ni–NTA affinity chromatography. The protein SoRIP1 reserved its inherent biological activities like topological inactivation and N glycosidase activity when tested and exhibited hemaglutination activity as well. SoRIP1 demonstrated cytotoxicity against a liver carcinoma cell line (HepG-2) and human breast adenocarcinoma cell line (MCF-7). SoRIP1 protein when tested against pathogenic bacterial (Ralstonia solanaceum and Acinetobacter lwoffii) and fungal (Alternaria solani and Alternaria Alternata) strains demonstrated growth inhibiting effect. These results suggested that SoRIP1 may be a potential anticancer and antimicrobial agent to be used in medicinal and agricultural field.