The continues and tireless researches on the development of biodiesel is still going on because of the demand and the expectation of the biodiesel is not meet out to replace the environmental hazardous fossil fuel. India the country with rich flora and fauna and great geographical topology for growing all types of plants and with 80% of the population practicing agriculture for their livelihood has not satisfactorily meet out the requirements of biodiesel production. The researches were repeatedly carried out in limited biodiesel feedstock and new feed-stocks are neither tried nor tried in a reluctant manner. If studies of biodiesel with all available plant oils were carried out extensively and exclusively, the rural employability and the production would have been boomed up and if found suitable for replacing diesel the demand to supply gap will be filled. In this experimental study the biodiesel was prepared from Cleome viscosa plant oil a new feedstock for biodiesel production which is a seasonal plant growing almost in all parts of India. The biodiesel produced from Cleome viscosa methyl ester (CVME) was subjected to various testing in conventional direct injection CI engine using different blends B100, B80, B60, B40, B20 and diesel for performance, combustion and emission analysis to prove its suitability for substituting diesel. It has been observed that a reduction in green house gases like CO and HC to an extent of 30% and 20% respectively, but the BSFC was increased by 5.4% and a reduction in BTE of 4.7% was noted. The biodegradability with the environmental sustainable nature of Cleome viscosa will prefers it as a suitable biodiesel to substitute diesel.