Translated Titles

Application of chitin particles on the purification of chitinolytic enzymes and proteases





Key Words

蛋白酶 ; 幾丁質酶 ; 磁性幾丁聚醣/四氧化三鐵納米複合顆粒 ; 粗製蝦殼幾丁質 ; Protease ; chitinase ; Magnetic chitosan / Fe3O4 nanocomposite particles ; Crude SSC



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Chinese Abstract

使用幾丁類物質粗製蝦殼粉 (Crude SSC) 與磁性幾丁聚醣/四氧化三鐵納米複合顆粒 (NF) 以吸附方式純化酵素,以達成快速純化之目的。 由吸附劑SP、AC、pure chitin、Crude SSC、Crude CSC、Crude SPC中選擇較適吸附劑Crude SSC。再探討溫度、重量、時間對吸附鳳梨酵素之影響。經由實驗得知於4℃、30分鐘、0.1 g的條件下,得到比活性0.0918 U/mg。 再探討溫度、重量、時間對NF吸附鳳梨酵素之影響。從實驗得知於25℃、30分鐘、0.1 g 的條件,得到比活性0.1357 U/mg。 使用上述條件,使用NF與Crude SSC吸附細菌酵素,藉此討論吸附方式純化酵素的結果。經由實驗可知純化酵素影響條件並非只有等電點的因素,也需考慮酵素和吸附劑的相互作用還有物理吸附等因素。 結果證明吸附方式純化酵素是一個可行的純化方式,可以提高純化效率。

English Abstract

Application of NF and Crude SSC on the purification of enzymes to achieve rapid purification purposes. Adsorbent : SP, AC, pure chitin, Crude SSC, Crude CSC, Crude SPC , Choose a more suitable adsorbent Crude SSC. Studying temperature, weight, time on the adsorption effect of the pineapple enzyme. By experiment that at 4 ℃, 30 min, 0.1 g of the conditions to get specific activity 0.0918 U / mg. Studying temperature, weight, time on the effect of NF adsorption bromelain enzyme. Learned from the experiment at 25 ℃, 30 min, 0.1 g of the conditions to get specific activity 0.1357 U / mg. Use the above conditions, NF, and Crude SSC absorption bacterial enzyme purified enzyme to discuss the results. Purified enzyme affected by the experimental conditions known isoelectric point is not the only factor, but also need to consider the interaction between enzyme and adsorbents as well as physical absorption and other factors. Results show that absorption is a viable way to purify the enzymes, it can improve the purification efficiency.

Topic Category 基礎與應用科學 > 化學
理學院 > 化學學系碩士班
  1. Alexandre-Franco M, Fernández-González C , Alfaro-Domínguez M, Gómez-Serrano V (2011) Adsorption of cadmium on carbonaceous adsorbents developed from used tire rubber. Journal of Environmental Management ,92 :2193-2200.
  2. Dotto G L, Pinto L A A (2011) Adsorption of food dyes onto chitosan: Optimization process and kinetic. Carbohydrate Polymers, 84 :231–238.
  3. Fajardo P, Martins J T , Fucinos C , Pastrana L, Teixeira J A, Vicente A A (2010) Evaluation of a chitosan-based edible film as carrier of natamycin to improve the storability of Saloio cheese . Journal of Food Engineering, 101 : 349–356
  4. Gupta B, Arorab A, Saxenaa S , Alam M S (2008) Preparation of chitosan–polyethylene glycol coated cotton membranes for wound dressings: preparation and characterization, Polymers for Advanced Technologies, 20: 58–65.
  5. Harish Prashanth K V ,Tharanathan R N (2007) Chitin/chitosan: modifications and their unlimited Application potentialdan overview. Trends in Food Science & Technology 18:117e131
  6. Hena S (2010) Removal of chromium hexavalent ion from aqueous solutions using biopolymer chitosan coated with poly 3-methyl thiophene polymer. Journal of Hazardous Materials, 181 :474–479
  7. Huang H Y, Shieh Y T, Shih C M, Twu Y K (2010) Magnetic chitosan/iron (II, III) oxide nanoparticles prepared by spray-drying . Carbohydrate Polymers ,81:906–910
  8. Huang X Y, Bin J P, BuH T, Jiang G B , Zeng M H (2011) Removal of anionic dye eosin Y from aqueous solution using ethylenediamine modified chitosan. Carbohydrate Polymers ,84:1350–1356
  9. Jellouli K, Ghorbel-Bellaaj O, Ayed H B, Manni L, Agrebi R, Nasri M (2011) Alkaline-protease from Bacillus licheniformis MP1: Purification, characterization and potential application as a detergent additive and for shrimp waste deproteinization. Process Biochemistry , 46 : 1248–1256.
  10. Kao P M , Chen C I , Huang S C, Lin K M, Chang Y C, Liu Y C (2009) Preparation of fermentation-processed chitin and its application in chitinase affinity adsorption. Process Biochemistry ,44 : 343–348
  11. Li J, Bai R (2002) Mechanisms of lead adsorption on chitosan/PVA hydrogel beads. Langmuir, 18: 9765–9770.
  12. Lu L C, Wang C I, Sye W F (2011) Applications of chitosan beads and porous crab shell powder for the removal of 17 organochlorine pesticides (OCPs) in water solution. Carbohydrate Polymers , 83:1984–1989.
  13. Reese T A, Liang H E, Tager A M, Luster A D, Van Rooijen N, Voehringer D, Locksley R M (2007) Chitin induces accumulation in tissue of innate immune cells associated with allergy. Nature, 447: 92–96.
  14. Kuroiwa T, Noguchi Y, Nakajima M , Sato S , Mukataka S, Ichikawa S (2008) Production of chitosan oligosaccharides using chitosanase immobilized on amylose-coated magnetic nanoparticles . Process Biochemistry 43: 62–69
  15. Pinto P X, Al-Abedb S R , Reismanb D J (2011) Biosorption of heavy metals from mining influenced water onto chitin products. Chemical Engineering Journal ,166 : 1002–1009
  16. Vijayaraghavan K ,Joshi U M , Balasubramanian R (2010) Removal of Metal Ions from Storm-Water Runoff by Low-Cost Sorbents: Batch and Column Studies. Journal of Environmental Engineering , 136:1113-1119
  17. Wang S L, Chen S J, Wang C L (2008b) Purification and characterization of chitinases and chitosanases from a new species strain Pseudomonas sp. TKU015 using shrimp shells as a substrate. Carbohydrate Research, 343:1171-1179.
  18. Wang S L, Pen J H, Liang T W, Liu K C (2008a) Purification and characterization of a chitosanase from Serratia marcescens TKU011. Carbohydrate Research, 343: 1316–1323
  19. Wang S L, Shieh J L, Liang T W (2011) Biodegradation of shellfish wastes and production of chitosanases by a squid pen-assimilating bacterium, Acinetobacter calcoaceticus TKU024. Biodegradation, pp.
  20. Zhang Y , Venugopal J R , El-Turki A , Ramakrishna S, Su B , Lim C T (2008) Electrospun biomimetic nanocomposite nanofibers of hydroxyapatite/chitosan for bone tissue engineering . Biomaterials , 29 :4314–4322
  21. 謝佳霖(2010) Bacills cereus TKU022 所生產幾丁聚醣酶及蛋白酶之特性分析與應用。淡江大學生命科學研究所碩士班碩士論文。
  22. Linden J C, Stoner R J, Knutson K W, Gardner-Hughes C A (2000) Organic disease control elicitors. Agro Food Industry Hi-Tech, 5: 32-34.