Extraction of Tannase by the New Strain of Penicillium

Title:Extraction of Tannase by the New Strain of Penicillium

Volume: 6 Issue: 4

Author(s): Roberta Cruz, Julyanna C. Fonseca, Marília de H. C. Maciel, Juliana S. de Lima, Minelli Albuquerque Sousa, Polyanna N. Herculano, Gladstone Alves da Silva, Erika V. Medeiros, Cristina Souza-Motta and Keila A. Moreira*

Affiliation:

• Keila A. Moreira. Academic Unit of Garanhuns, Federal Rural University of Pernambuco, Garanhuns - PE, 55292-270,Brazil

Keywords: Aqueous two-phase system, tannase, purification factor, enzyme characterization, Penicillium rolfssi, strain.

Abstract: Backgrond: The aqueous two-phase systems (ATPS) have been used as the first purification step in enzyme industrial isolation. Such systems allow the removal of large amounts and different types of contaminants by a simple and economic unit operation.

Objective: The tannase extraction was conducted through ATPS composed of polyethylene glycol (PEG) and sodium citrate salt (PEG/citrate).

Method: A factorial design model (24) was used to assess the influence of PEG molar mass (400-8000 g mol−1 - MPEG), PEG concentration (12.5-17.5%, w/w - CPEG), citrate concentration (15-25%, w/w, CCIT) and pH (6.0-8.0) on tannase extraction.

Results: The tannase is preferably partitioned at the top phase. The highest purification factor (PF = 18.18) was achieved in ATPS using 25% (w/w) PEG 8000 (g mol−1) and 25% (w/w) CCIT at pH 8.0, in which a yield (Y) of 23.37% was obtained. Furthermore, pH was the independent variable that most significantly influenced the response variables. Tannase showed pH and optimum temperature of 5.0 and 60°C, respectively. It was stable in a very short pH range (3.0-6.0) and unstable in the studied temperature range (25-70°C). The tannase activity was stimulated by Ca2+, Co2+, Fe2+ and Mn2+. Organic solvents also increased enzyme activity, particularly in the following rates: 40% chloroform, 20% and 40%, glycerol and 20% and 40% toluene.

Conclusion: The ATPS could be used as the first step in tannase extraction from a crude extract, and as a promising and low cost alternative for partial purification of tannase produced by P. rolfssi URM 6216.

Cite this article as:

Cruz Roberta, Fonseca C. Julyanna, de H. C. Maciel Marília, de Lima S. Juliana, Albuquerque Sousa Minelli, Herculano N. Polyanna, da Silva Alves Gladstone, Medeiros V. Erika, Souza-Motta Cristina and Moreira A. Keila*, Extraction of Tannase by the New Strain of Penicillium, Current Biotechnology 2017; 6 (4) . https://dx.doi.org/10.2174/2211550105666160316004323