The Tannase from red yeast Rhodotorula glutinis: purification and characterization

Abstract

The tannase enzyme was successfully purified to homogeneity from the culture broth of red yeast strain Rhodotorula glutinis DB2 in a three-tandem step involving ultrafiltration (5 kDa and 100 kDa systems), Sephadex G-200 gel filtration chromatography, and DEAE Sepharose CL-4B anion exchange chromatography. The purified tannase appeared to be homogeneous on sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS-PAGE). The purified tannase had a specific activity of 3.33 U mg⁻¹, with a 1.3% recovery and overall purification of 302-fold. The molecular mass of the tannase estimated by SDS-PAGE was about 73 kDa. The tannase had an optimum pH of 6.0 and an optimum temperature of 40 °C. The most stable pH was 7.0, and the enzyme was stable up to 40 °C. One mmol L⁻¹ of Fe³⁺, Sr²⁺, Na⁺, and Pb²⁺ were found to promote tannase activity, whilst 1.0 mmol L⁻¹ of Ba²⁺, Ca²⁺, Mg²⁺, Zn²⁺, Hg⁺, Ag⁺, Co²⁺, Fe²⁺, Mn²⁺, Cu²⁺, Cd²+, Al³⁺, K⁺, Ni²⁺, and Li⁺ inhibited tannase activity.

Graphical Abstract

Keywords:

• Tannase
• Rhodotorula glutinis
• red yeast
• purification
• characterization

Acknowledgments

The authors would like to thank the Universiti Sains Malaysia for the financial support to conduct this research.

Ethics approval

Not applicable.

Consent to participate

Not applicable.

Author contributions

Methodology: CBO, and DI; Conceptualization: CBO, DI, and MJNMK; Formal analysis: CBO, and DI; Funding acquisition: DI; Investigation: CBO; Writing original draft: CBO; Review and Editing: CBO, and DI.

Disclosure statement

No potential conflict of interest was reported by the author(s).