A CONTRIBUTION TO THE CHARACTERIZATION OF PROTOPECTINASE SE, AN ENDOPOLYGALACTURONASE WITH PECTIN-RELEASING ACTIVITY FROM Geotrichum klebahnii

  • S.F. Cavalitto Research and Development Center for Industrial Fermentation (CINDEFI; UNLP, CONICET La Plata), School of Science, La Plata National University, 47 y 115 (B1900ASH)
  • R.A. Hours Research and Development Center for Industrial Fermentation (CINDEFI; UNLP, CONICET La Plata), School of Science, La Plata National University, 47 y 115 (B1900ASH)
  • C.F. Mignone Research and Development Center for Industrial Fermentation (CINDEFI; UNLP, CONICET La Plata), School of Science, La Plata National University, 47 y 115 (B1900ASH)
Keywords: endo-polygalacturonase, kinetic characterization, metal inhibition

Abstract

Hydrolysis of polygalacturonic acid (1.8 g.L−1 ) with 1.25 mg.L−1 of a commercial preparation (Pectinase SE from Shikibo Ltd., Japan) containing protopectinase SE (PPase-SE) activity for 80 min released a reducing power equivalent to ≈ 400 mg.L−1 of galacturonic acid monohydrate, yielding oligogalacturonates with an average polymerization degree of around 4. Thermostability of PPase-SE was positively affected by enzyme concentration. Enzyme activity of a solution containing 12.5 mg.L−1 of Pectinase SE in 20 mM sodium acetate buffer, pH 5.0, quickly dropped to less than 20 % after 80 s of vortexing. Enzyme pre-incubation (37°C, 30 min) with Ca2+, Mg2+, Co2+, Cu2+, Fe2+, Zn2+ and Mn2+ (0.1, 1.0 and 10.0 mM) did not affect the activity except in the case of Cu2+ (10 mM). Ca2+ (2.5 mM) and Mg2+ (2.5 mM) in the reaction mixture caused inhibition and activation, respectively. Ca2+ inhibition was partially reverted by Mg2+. The enzyme is not inhibited by products. Km and Vmax value for PGA was determined to be 0.198 ± 0.013 g.L−1 and 0.0509 ± 0.0032 µmol.mL−1 .min−1 respectively. This Km value is one of the lowest reported for microbial PGases from different origins.

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Published
2020-02-07
How to Cite
Cavalitto, S., Hours, R., & Mignone, C. (2020). A CONTRIBUTION TO THE CHARACTERIZATION OF PROTOPECTINASE SE, AN ENDOPOLYGALACTURONASE WITH PECTIN-RELEASING ACTIVITY FROM Geotrichum klebahnii. Revista Mexicana De Ingeniería Química, 13(1), 75-81. Retrieved from http://rmiq.org/ojs311/index.php/rmiq/article/view/1298
Section
Biotechnology