Purification and characterization of two cold-adapted extracellular tannin acyl hydrolases from an Antarctic strain Verticillium sp. P9

• Published in: Applied microbiology and biotechnology Vol. 77; no. 1; pp. 77 - 89
• Main Authors: Kasieczka-Burnecka, Monika, Kuc, Karina, Kalinowska, Halina, Knap, Monika, Turkiewicz, Marianna
• Format: Journal Article
• Language: English
• Published: Berlin Berlin/Heidelberg : Springer-Verlag 01-11-2007; Springer; Springer Nature B.V
• Subjects: Acids; Adaptation, Physiological; Anions; Antarctic cold-adapted extracellular tannases; Antarctic Regions; Ascomycota - enzymology; Ascomycota - growth & development; Biochemistry; Biological and medical sciences; Biology of microorganisms of confirmed or potential industrial interest; Biotechnology; Carbohydrates; Carboxylic Ester Hydrolases - chemistry; Carboxylic Ester Hydrolases - isolation & purification; Carboxylic Ester Hydrolases - metabolism; Catalysis - drug effects; Cellulose; Chelating Agents - pharmacology; Chromatography; Chromatography, DEAE-Cellulose; Cold; Cold Temperature; Copper; Dehydrogenases; Dose-Response Relationship, Drug; Electrophoresis, Polyacrylamide Gel; Enzyme Stability - drug effects; Enzymes; Fundamental and applied biological sciences. Psychology; Fungal Proteins - chemistry; Fungal Proteins - isolation & purification; Fungal Proteins - metabolism; Fungi; Glycoproteins; Isoenzymes - chemistry; Isoenzymes - isolation & purification; Isoenzymes - metabolism; Kinetics; Metals - pharmacology; Miscellaneous; Mission oriented research; Molecular Weight; Proteins; Protons; Sodium Chloride - pharmacology; Surface-Active Agents - pharmacology; Tannic acid; Tannins; Tea; Temperature; Thermodynamics; Urea; Urea - pharmacology; Verticillium sp; Antarctica; Antarctic Regions; Fungi; Characterization; Purification; Enzyme; Tannase; Verticillium; Hydrolases; Esterases; Fungi Imperfecti; Extracellular; Physicochemical properties; Carboxylic ester hydrolases
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s00253-007-1124-4

Two extracellular tannin acyl hydrolases (TAH I and TAH II) produced by an Antarctic filamentous fungus Verticillium sp. P9 were purified to homogeneity (7.9- and 10.5-fold with a yield of 1.6 and 0.9%, respectively) and characterized. TAH I and TAH II are multimeric (each consisting of approximately 40 and 46 kDa sub-units) glycoproteins containing 11 and 26% carbohydrates, respectively, and their molecular mass is approximately 155 kDa. TAH I and TAH II are optimally active at pH of 5.5 and 25 and 20°C, respectively. Both the enzymes were activated by Mg²⁺and Br- ions and 0.5-2.0 M urea and inhibited by other metal ions (Zn²⁺, Cu²⁺, K⁺, Cd²⁺, Ag⁺, Fe³⁺, Mn²⁺, Co²⁺, Hg²⁺, Pb²⁺ and Sn²⁺), [graphic removed] anions, Tween 20, Tween 60, Tween 80, Triton X-100, sodium dodecyl sulphate, β-mercaptoethanol, α-glutathione and 4-chloromercuribenzoate. Both tannases more efficiently hydrolyzed tannic acid than methyl gallate. E a of these reactions and temperature dependence (at 0-30°C) of k cat, k cat/K m, ΔG*, ΔH* and ΔS* for both the enzymes and substrates were determined. The k cat and k cat/K m values (for both the substrates) were considerably higher for the combined preparation of TAH I and TAH II.