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In the current study, three types of common plants, namely Tomato (Solanum lycopersicum), Cucumber (Cucumis sativus) and Orange (Citrus sinensis) were obtained and screened for their polyphenol oxidase (PPO) activity, Among the three plants, Solanum lycopersicum was chosen with maximum enzymatic activity, it had the highest productivity of the enzyme (23733 U/mg protein).The PPO from Solanum lycopersicum was purified using two steps: concentration by sucrose and gel filtration by using Sephacryl S-200. The results showed an increase in the final purification folds by 2.4 times with an enzyme yield of 32.6%. The immobilization studies showed that PPO was more stable when immobilized on chitosan by covalent linkage with immobilization ratio of 62%, in comparison with agar-agar by entrapment method (36%). The removal efficiency of crude and partial purified PPO was studied with textile dyes, including yellow, red, black and blue dyes at optimum conditions: pH 5, temperature 40 o C after 3 hrs. Maximum removal efficiency of dyes observed with crude PPO were 53.9, 81.4, 86.5 and 79.6% respectively. However, purified PPO displayed removal efficiency reached 60.3, 84.3, 84.6 and 77.5% respectively. The potential of immobilized PPO on chitosan was evaluated by decolorization of black textile dye in packed bed bioreactor in batch operation. The results indicated that immobilized PPO in batch operation has the ability to remove 99% of the dye after 2 hrs, and the results showed a positive relationship between the degradation rate and incubation time in batch operation. Keyword: Enzymes, Tomato Plants, Bioreactor, Industrial Dyes ةيقارعلا ةيعارزلا مولعلا ةلجم 2019 : 50 ) 3 ) : 943 950 نيسح ةءافك يويحلا لعافملا يف هبجولا ماظن مادختساب ةجسنلاا تاغبص ضعب كيكفت يف ديقملا زيديسكوا لونيف يلوبلا ميزنا نيسح ميحرا رحس سردم يئايحلاا تاينقتلا مسق ة قا رعلا/دادغب ةعماج/مولعلا هيلك saharraheem2015@gmail.com صلختسملا هيلاحلا هساردلا تنمضت هلبرغ تاتابن ةثلاث نلا اهجاتنا هيلباق يف ميز polyphenol oxidase ةطامطلا تلمش (Solanum lycopersicum) و رايخلا ( (Cucumis sativus و لاقتربلا (Citrus sinensis) هطامطلا تابن بختنا هيميزنا هيلاعف ىلعا ىلع هءاوتحاب تغلب ذا 23733 /ةدحو ميزنا يقن .نيتورب مغلم polyphenol oxidase نم جتنملا بختنملا تابنلا حيشرتلاب تناكف هيناثلا اما زوركسلاب زيكرتلا ىلولاا تنمضت نيتوطخب ليركافيسلا مادختساب يملاهلا s 200 هيقنتلا تا رم ددع يف ةدايز جئاتنلا تنيب ، هريخلاا هوطخلا يف تغلب ذا 2.4 ةيميزنلاا هليصحلاو 32.6 .% ديق ميزنلاا يرطب ناك ثيح نيتق ىلا دييقتلا هبسن تلصو اذا يمهاستلا طبرلا هطاسوب ناسوتيكلا هدامب هدييقت دنع تابث رثكا 62 هنراقم % ب هطاسوب هدييقت ا هدامب زجحلا تلصو اذا راكا راكلا ىلا 36 لا ميزنلاا هءافك تسرد .% ماخ لا ، ءا رفصلا هغبصلاب هلثمتملا هجسنلاا لماعم تاغبص هلا زا يف ىقنملاو ،ءا رمح هضومح هلادب هلثمتملا ىلثملا فورظلا دنع ءاقرزلاو ءادوسلا 5 ةرا رح هجردو 40 م o تاعاس ثلاث دعب هعبرلاا تاغبصلا هلزلا هءافك ىلعا تلصو . ىلا ماخلا ميزنلاا مادختساب 53.9 و 81.4 و 86.5 و %79.6 ىلا ىقنملا ميزنلاا مدختساب تاغبصلا هلا زلا هءافك ىلعا تلصو كلذك ، يلاوتلا ىلع 60.3 و 84.3 و 84.6 و 77.5 ءادوسلا هغبصلا هلازا يف ناسوتيكلا هدامب ديقملا ميزنلاا هءافك تسرد .يلاوتلا ىلع % يف هبجولا ماظنب لعافملا تغلب اذا ، يويحلا 99 حلا هسا ردلا تنيب .نيتعاس دعب % هيدرط هقلاع كانه هيلا هناضحلا ةرتفو هغبصلا هلا زا لدعم نيب هغبصلا هلازا هءافك يف ماظنب ةيجولا . :ةيحاتفم تاملك ميزنا ، تابن ةيعانص تاغبص ،يويح لعافم ،ةطامطلا *Received:12/7/2018, Accepted:1/11/2018 Iraqi Journal of Agricultural Sciences –2019:50(3):943950 Hussein 944 INTRODUCTION In line with the evolution of the petroleum industry, the area of oil-polluted soil has increased continuously. Thus, it becomes a natural question of treating the oil-polluted soil to retrieval level in a short time (30). The bioremediation technology could be beneficial because of its pollution-free, low cost and highly efficient characteristics. In recent years, this technology has developed quickly and different microorganisms with special biochemical and physiological functions have been screened, which promote the bioremediation technology for further application and development (12). The crude oil filtrated into soil through varying ways and interacted with soil enzymes (2). On the one hand, the enzymes from oil-degrading plants could be an indicator for assaying the amount and physiochemical property of crude oil. On the other hand, the enzymes had the great ability to degrade the petroleum. The activities of the enzymes indicated the intensity and tendency of various biochemical reactions, though the amounts of the enzymes were quite small. In a sense, the enzyme, a living molecular unified with the environmental factors, could degrade the oil to simple chemical compounds. It is necessary to give greater attention to investigating the relationship between environmental pollution and the enzymes from oil-degrading plants and the suitable parameters of enzyme activity. Polyphenol oxidase (PPO) is a monophenol, dihydroxy-L-phenylalanine oxygen oxidoreductase, (E.C.1.14.18.1), which occur in many plants together with phenolics. It’s an enzyme that catalyzes the oxidation of mono-, diand poly-hydric phenols to o-quinones. Different names have been associated with this enzyme including catecholase, cresolase, tyrosinase, phenolase and diphenolase (21). The enzyme is widely distributed on the phylogenic scale and is responsible for the harmful effect of enzymatic browning reactions in foods and other materials. It is known that browning reactions, initiated by the enzyme, occur generally post-harvest when the tissues are exposed to stress conditions, deterioration, or during handling, storage and processing (29). PPO has been widely studied and characterized in a variety of tissues, such as tomato (26), sweet potato (20), potato and taro (10), eggplant (7), palmito (20), apple (22), pears (13), grapes (24), peppermint (20), oregano (10) and artichoke (1). The aim of this study is the extraction, purification, immobilization and decolorization of some textile dyes by polyphenol oxidase from plant sources. MATERIALS AND METHODS Plants The plants used throughout this study were locally available in the market. Solanum lycopersicum (Tomato), Cucumis sativus (Cucumber) and Citrus sinensis (Orange) were used as a source of material to screen for polyphenol oxidase activity. Extraction and recovery of PPO One hundred gm of each plant were homogenized, with the mixing performed by using a blender for 10 mins at room temperature. The slurry was filtrated through gauze to remove any cell debris from the preparation, then centrifuged at 8000 rpm for 10 mins. The clear supernatant obtained represented the crude extract, and was assayed for PPO activity. PPO activity Polyphenol oxidase enzyme activity was estimated according to the method described by Zian and Pekyardimici (33) using orthocatechol as a substrate. The oxidation of orthocatechol was detected by measuring the absorbance increase at 420 nm using a VISspectrophotometer. The protein concentration was measured according to the method described by Bradford (4). Purification of PPO Purification of the enzyme was determined by two methods: concentration by dialysis and gel filtration. Enzyme concentration by dialysis The crude enzyme solution was concentrated by using sucrose, then the enzyme activity and the protein concentration was measured. Gel filtration chromatography The column was prepared and packed according to the instructions of the manufacturing company (Pharmacia-Sweden). The concentrated enzyme by dialysis was passed through sephacryle S-200 column with the dimensions 21×1.6 cm. The elution was done by using 0.2M of sodium phosphate Iraqi Journal of Agricultural Sciences –2019:50(3):943950 Hussein 945 buffer solution, pH 7.0 with flow rate of 20 ml / h, 3ml for each fraction. The protein was estimated in each fraction at wave length 280 nm, then enzyme activity was measured in these fractions. Effective fractions were collected and then its volume was measured as well as its activity and protein concentration were estimated. Immobilization of PPO enzyme Immobilization of PPO by entrapment

EFFECIENCY OF IMMOBILIZED POLYPHENOL OXIDASE ON SOME TEXTILE DYES DEGRADATION USING BATCH OPERATION SYSTEM BY PACKED BED BIOREACTOR