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Volume: 7, Issue: 1, Jan-Feb, 2019
DOI: 10.7324/JABB.2019.70106

Research Article

Purification and characterization of tannase from the local isolate of Aspergillus niger

Saja Taha Yaseen Al-Mraai, Dhia Falih Al-Fekaiki, Alaa Jabbar Abd Al-Manhel

  Author Affiliations


Fungi of genus Aspergillus are active producers of extracellular tannase, and characteristics of enzyme production by local Aspergillus species have been well studied. The aim of this experimental study is isolation and partial purification of tannase enzyme from the Aspergillus niger fungus species. The purification was achieved by applying respectively ammonium sulfate (50-70%), dialysis with a specific activity 1227.08 (unit/mg protein), then passed the enzymatic extract through superdex 200 column in the gel filtration chromatography by using ÄKTA Pure 25 apparatus, the activity and the specific activity reached to 525.12 (unit/ml) and 2917.33 (unit/mg protein) respectively, with purification fold of 11.629 and yield 18.40%. One peak was obtained from an enzyme purity, which diagnosed by the absence of denaturation substances for protein SDS. Characteristics of pure enzyme showed that the molecular weight of the pure enzyme was determined, and it was found that the weight of enzyme was 95.49 KDa by using sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis, the enzyme was active in pH range 3 to 6 and temperature of 20 to 50°C. The optimum pH and the temperature for tannase activity were recorded to be pH 5 and 35°C, respectively. Moreover, the tannase activity was inhibited by Cu, Fe and Hg ions while increased by Mg and K ions when added at 1 and 5 mM.


Fungi, Aspergillus niger, Tannase, Purification, Characterization.

Citation: Al-Mraai STY, Al-Fekaiki DF, Abd Al-Manhel AJ. Purification and characterization of tannase from the local isolate of Aspergillus niger. J App Biol Biotech. 2019;7(01):029-034. DOI: 10.7324/JABB.2019.70106

Copyright: Author(s). This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License, which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited.


1. Girdhari S, Peshwe S. Isolation and Screening of Tannase producing fungi. International Journal of Current Microbiology and Applied Sciences 2015; 4:765-774.

2. Kumari M, Jani S. Tannins: Antinutrient with positive effect to Manage diabetes. Journal of Recent Sciences 2012; 1:70-73.

3. Aguilar-Zárate P, Cruz-Hernández M, Montanez J, Belmares-Cerda R. Aguilar C. Bacterial Tannases: Poduction, Properties and Applications Tanasas bacterianas: Produccion, Propiedadesy Aplicacioness. Revista Mexicana de Ingeniería Química 2014; 13:63-74.

4. Gemede HF, Ratta N. Antinutritional factors in plant foods: potential health benefits and adverse effect. International Journal of Nutrition and Food Sciences 2014; 3:284-289. https://doi.org/10.11648/j.ijnfs.20140304.18

5. Abd El Tawab AM, Khattab MSA. Utilization of polyethylene glycol and tannase enzyme to reduce the negative effect of tannins on digestibility, milk production and animal performance. Asian Journal of Animal and Veterinary Advances 2018; 13:201-209. https://doi.org/10.3923/ajava.2018.201.209

6. Mcgiinnis MR. 1980. Laboratory handbook of medical mycology. New York: Academic Press, Inc.

7. Pitt JI, Hocking AD. Fungi and food spoilage: 3rd Edition, Springer US; 2009; XV,519. https://doi.org/10.1007/978-0-387-92207-2

8. Al-Mraai STY, Al-Fekaiki DF, Al-Manhel AJA. Study the optimal conditions for the production of tannase from local Aspergillus niger by solid state fermentation. Misan Journal of Academic Studies 2018, in press.

9. Sharma NK, Beniwal V, Kumar N, Kumar S, Pathera AK, Ray A. Production of tannase under solid state fermentation and its application in detannification of guava juice. Preparative Biochemistry and Biotechnology 2014; 44:281-290. https://doi.org/10.1080/10826068.2013.812566

10. Mondal K, Banerjee D, Jana M, Pati B. Colorimetric assay method for determination of the tannin acyl hydrolase (EC 3.1. 1.20) actvity. Analytical Biochemistry 2001; 295:168-171. https://doi.org/10.1006/abio.2001.5185

11. Lowry OH, Rosobrough N, Far AL, Randall RJ. Protein measurement with Folin phenol reagent. Journal of Biological Chemistry 1951; 193:265-275.

12. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature Publishing Group 1970; 227:680- 685. https://doi.org/10.1038/227680a0

13. Garfin DE. 1990. Purification procedures electrophoretic methods. In: Murray ED, Dentscher PJ, editors. Methods in Enzymology. New York: Academic Press 425-441.

14. Liu TP, Porto TS, Moreira KA, Takaki GM, Brand\ão-Costa R, Herculano PN, Porto AL. Tannase production by Aspergillus spp. UCP1284 using cashew bagasse under solid state fermentation. African Journal of Microbiology Research 2016; 10:565-571. https://doi.org/10.5897/AJMR2016.7924

15. Cavalcanti RMF, de Oliveira Ornela PH, Jorge JA, Souza LH. Screening, Selection and Optimization of the Culture Conditions for Tannase Production by Endophytic Fungi Isolated from Caatinga. Journal of Applied Biology & Biotechnology 2017; 5:001-009.

16. Philip DC, Lavanya B, Latha S. Purification of tannase from Aspergillus niger under Solid State fermentation. World Journal of Pharmacy and Pharmaceutical Sciences 2015; 4:993-1001.

17. Nandi S, Chatterjee A. Extraction, partial purification and application of tannase from Aspergillus niger MTCC 2425. International Journal of Food Science and Nutrition 2016; 1:20-23.

18. Anitha A, Arunkumar D. Extraction, Partial purification and characterization of tannase enzyme from Mucor sp. Nehru E-Journal 2013; 1:25-29.

19. Mahendran B, Raman N, Kim DJ. Purification and characterization of tannase from Paecilomyces variotii: hydrolysis of tannic acid using immobilized tannase. Applied Microbiology and Biotechnology 2006; 70:444-450. https://doi.org/10.1007/s00253-005-0082-y

20. Viswanath V, Leo VV, Prabha, SS, Prabhakumari C, Potty V, Jisha M. Biosynthesis of tannase from cashew testa using Aspergillus niger MTCC5889 by solid state fermentation. Journal of Food Science and Technology 2015; 52:7433-7440. https://doi.org/10.1007/s13197-015-1858-4

21. Enemuor SC, Odibo FJC. Partial purification and characterization of Aspergillus tamarii IMI388810 (B) tannin acyl hydrolase. Archives of Applied Science Research 2010; 2:290-299.

22. Wagh SA. Bioconversion of tannic acid to gallic acid by using fungal tannase. Ph.D. thesis, CSIR-National Chemical Laboratory, Pune, India; 2010.

23. El-Tanash A, Sherief A, Nour A. Catalytic properties of immobilized tannase produced from Aspergillus aculeatus compared with the free enzyme. Brazilian Journal of Chemical Engineering 2011; 28:381-391. https://doi.org/10.1590/S0104-66322011000300004

24. Ordo-ez R, Colombo I, Alberto M, Isla M. Production of tannase from wood-degrading fungus using as substrate plant residues: purification and characterization. World Journal of Microbiology and Biotechnology 2011; 27:2325-2333. https://doi.org/10.1007/s11274-011-0699-1

25. Whitaker JR. 1972. Principles of enzymology for the food science. New York, USA: Marcel Dekker. Inc.

26. Bagga J, Pramanik SK, Pandey V. Production and Purification of Tannase from Aspergillus aculeatus Using Plant Derived Raw Tannin. International Journal of Scientific Engineering and Technology 2015; 2:50-55. https://doi.org/10.17950/ijset/v4s2/205

27. da Costa AM, Kadowaki MK Minozzo MC, de Souza CGM, Boer CG, Bracht A, Peralta RM. Production, purification and characterization of tannase from Aspergillus tamarii. African Journal of Biotechnology 2012; 11:391-398.

28. Yao J, Guo GS, Ren GH, Liu YH. Production, characterization and application of tannase. Journal of Molecular Catalysis B: Enzymatic 2014; 101:137-147. https://doi.org/10.1016/j.molcatb.2013.11.018

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