Most of the Euphorbian plants secrete fluid which contain a proteolytic enzyme for defensive role against insects, pests and hence eco physiological inheritance to sustain vegetation eventually in adverse environmental conditions. Evaluation has been carried out on twenty five Euphorbian garden plants for their proteolytic activities using casein as a substrate. Out of these, Euphorbia nerifolia, Euphorbia milli, Euphorbia tirucalli, Euphorbia lactea, Synadenium granti, Jatropha curcas, Euphorbia nivulia, Euphorbia antiquoram, Pedilanthus tithymaloides, Euphorbia viguieri, E. heterophylla and E. leucocephala are the good enzyme source. Moderate activity found in Jatropha integerrima, Jatropha multifida, Jatropha podagrica, Euphorbia pulcherrima, and Dalechampia scandens. While different tissues of Acalypha hispida, Acalypha wilkesiana, Breynia nivosa, Cicca acida, Codiaeum variegatum, Drypetes roxburghii are devoid of proteolytic activity. This paper describes in detail about name of plants, habitat and presence of proteolytic enzyme in them. Results show that the out of twenty five plants 50% plant tissue synthesize protease in appreciable amount, while 10% are not able to produce it. However 40% plants demonstrate only detectable amount of protease. A comparative account of proteolytic activity reveals some promising plants good source of enzyme. Some plants produce combination of cysteine and serine proteases. A single plant i.e. Euphorbia nerifolia latex contains cysteine, serine, metallo-protease and aspartic proteases. In turn, these proteases may be used in various industrial uses in general and cheese production in a particular. Key-wordsGarden Euphorbian plants, Cysteine and serine protease, E. leucocephala, Euphorbia viguieri -------------------------------------------------IJLSSR----------------------------------------------INTRODUCTION Botanical plants are considered as god’s gift to human being in the form of natural medicine. Euphorbiaceae comprises more than 2000 species. Some of Euphorbian plants are cultivated as ornamental/garden plants in national and international gardens. Due to rich cultural heritage and relatively rich flora, a wealth of knowledge on traditional and folk medicine has been accumulated in India [1]. An exhaustive and a comprehensive review on proteolytic enzyme of biological sources appeared in literature which includes study on properties of various proteases with mechanism of action of proteolysis of protein [2]. Access this article online Quick Response Code: Website: www.ijlssr.com DOI: 10.21276/ijlssr.2016.2.4.7 The used parts of Euphorbian plant species include latex, roots, seeds, stem bark, wood, leaves and whole plant [3-5]. The plants in the family Euphorbiaceae are known for chemical diversity of secondary metabolites and have various curative properties against different ailments [6]. Most of member of this family synthesis proteases in different tissues for defensive purpose [7-9]. A good source is latex and juices. Lynn described occurrence, properties of different proteases of Euphorbiaceae family [7]. An excellent article is appeared in literature stating importance of a chemotaxonomic marker of Euphorbia species pertaining presence of proteolytic activity in the latex of Euphorbian genera [9]. This aspect is confirmed recently in next year in the form of review of Euphorbiaceae family and its medicinal features [10]. Further such study is extended for the production of plant proteases in vivo and in vitro [11]. A scientific article on research into Euphorbia latex and various ingredients is published [12]. Very recently, article entitled a study on plant latex, a rich source of proteases and cutting edge for disease invasion is appeared in literature mentioning that, Research Article (Open access) Int. J. Life. Sci. Scienti. Res., VOL 2, ISSUE 4 http://ijlssr.com IJLSSR © 2015 All rights are reserved Page 356 out of the 35 latex bearing plants, 16 plants possess proteolytic activity belongs to family of Euphorbiaceae [13]. Very very recently, a very good article entails medicinal importance and biochemistry of latex of certain Euphorbian taxa [8]. In our laboratory, we mentioned 13 industrial bio-applications of proteases of some Euphorbian wild and weed plants [14]. In this communication, we report here studies on proteases in garden Euphorbian plants aiming to search a vegetable rennin source for production of cheese and some allied industrial applications. MATERIALS AND METHODS Plants are procured from campus of Moolji Jaitha College, Jalgaon and plant nurseries of Jalgaon city, Maharashtra, India. Dr. Tanveer taxonomist, identified plants for study. Different parts of plant such as leaf, stem, root, flower, and latex of the garden Euphorbian plants was collected from during June 2014 to December 2015. The cuttings of leaf stalks with capillary tubes into glass container and was kept in ice. The latex was a white thick fluid with pungent odour brought to laboratory and kept in refrigeration till use. Experiments were conducted at department of Biotechnology, PGCSTR, Jalgaon, India. A photo plate of some promising Euphorbian garden plants is given in Fig. 1. Fig. 1 Photo plates of 10 Euphorbian garden plants Enzyme Isolation The freshly collected latex was diluted with 5 volumes of ice cold phosphate buffer pH 7.4 and centrifuged at 10,000 rpm for 20 minutes in high speed refrigerated centrifuge and supernatant was collected and stored at 40C. The pellet containing the white insoluble gum was discarded. All the experiments on the crude preparation were carried out using freshly collected latex and preserved in refrigerator at 40C. From other parts 10% homogenate was prepared in phosphate buffer at pH 7.4 and centrifuged and supernatant was used as a source of enzyme. Int. J. Life. Sci. Scienti. Res., VOL 2, ISSUE 4 http://ijlssr.com IJLSSR © 2015 All rights are reserved Page 357 Screening and Selection of Garden Euphorbian Plant Proteases Protease Activity Proteolytic activity of different plant tissues was determined by the colorimetric assay using 1% casein as a substrate as described by [15]. The protease activity was expressed as amount of enzyme required to produce peptide equivalent to μg of tyrosine/min/mg protein at 370C and protein content was determined according to Lowry’s method [16] using Bovine serum albumin as the standard protein. Enzyme Unit One unit of protease activity is defined as the amount of enzyme to release 1 μg of tyrosine per minute at 370 C. A tyrosine standard curve was calibrated (10 to 100 μg/ml) using Folin Phenol reagent. Specific activity of the proteolytic enzyme is expressed as the number of units per milligram of protein. RESULTS AND DISCUSSION Proteases are distributed widely in different biological sources namely plants, animals and microbial sources. In Euphorbian plants protease are present in virtually every part i.e. stem, fruit, flower, leaf, root, gum and latex. We have communicated presence of proteolytic activity in various parts of plant indicated, plant latex is the richest source of protease [14,17]. Table 1 summarizes habitat of some Euphorbian plants. They are grouped into three category i. Wild, ii. Weed, and iii. Garden. The distribution of wild, weed and garden is 47%, 22%, and 31% respectively. The order of occurrence of protease in garden Euphorbian plant is serine< cysteine< serine and cysteine < metallo protease<aspartic protease (Fig. 2). This finding is good in agreement and supports our findings as results reported by number of authors [9-10, 18-19] pertaining to the occurrence and distribution of proteolytic enzymes. Table 1: List of Some Euphorbian Plants The life form of above plants ranging from small herbs, herbs, shrubs, small tree and tree. Some of them are seasonal and perennial. Wild (A) Weed (B) Garden (C) Acalypha ciliate Acalypha malabarica Acalypha hispida Acalypha indica Chrozophora prostrate Acalypha wilkesiana Baliospermum raziana Chrozophora rottleri Breynia nivosa Bridelia airy-shawii Euphorbia hirta Cicca acida Cleidion spiciflorum Euphorbia indica Codiaeum variegatum Croton bonplandianum Euphorbia notoptera Drypetes roxburghii Emblica officinalis Euphorbia prostrate Euphorbia milii Euphorbia clarkeana Euphorbia prunifolia Euphorbia pulcherrima Euphorbia cristata Euphorbia thymifolia Euphorbia tirucalli Euphorbia fusiformis Phyllanthus airy-shawii Jatropha integerrima Euphorbia nerifolia Phyllanthus amarus Jatropha multifida Euphorbia nivulia P. maderaspatensis Jatropha podagrica Euphorbia pycnostegia Pedilanthus tithymaloides Homonoia riparia Synadenium granti Jatropha curcas Dalechampia scandens Jatropha gossypifolia Euphorbia viguieri Kirganelia reticulate Jatropha podogrica Mallotus philippensis Euphorbia nerifolia Manihot esculenta Euphorbia nivulia Micrococca mercurialis Euphorbia species 1 Phyllanthus urinaria Euphorbia species 2