ORIGINAL ARTICLE   

Minerva Biotecnologica 2020 June;32(2):43-51

DOI: 10.23736/S1120-4826.20.02605-1

Copyright © 2020 EDIZIONI MINERVA MEDICA

language: English

Polymer impregnation in porous glass beads to induce bioseparation of β-mannanase from fermentation broth of Proteus vulgaris

Joo S. TAN ¹ ✉, Sahar ABBASILIASI ², Arbakariya B. ARIFF ³, Paramasivam MURUGAN ¹, Mohamad H. ABU BAKAR ¹, Hui S. NG ⁴

¹ Department of Bioprocess Technology, School of Industrial Technology, Sains Malaysia University, Pulau Pinang, Malaysia; ² Halal Products Research Institute, Putra Malaysia University, Selangor, Malaysia; ³ Bioprocessing and Biomanufacturing Research Center, Faculty of Biotechnology and Biomolecular Sciences, Putra Malaysia University, Selangor, Malaysia; ⁴ Department of Food Science and Nutrition, Faculty of Applied Sciences, UCSI University, Kuala Lumpur, Malaysia

BACKGROUND: Mannanase has been used for the hydrolysis of mannan, which is produced by many microorganisms including bacteria. The purified enzyme can be utilized in the production of high-value product such as mannooligosaccharides as prebiotics. This study aimed at purification of extracellular β-mannanase from Proteus vulgaris using an aqueous polymer impregnated resin system.
METHODS: The effect of extraction solution (potassium phosphate, sodium citrate and sodium acetate) on the stability of β-mannanase activity was studied. The parameters which influence the purification performance such as molecular weight and concentration of PEG, concentration of salts, pH of salt, crude loading and extraction time were investigated using one-factor-at-a-time (OFAT). The effect of type and concentration of salts and pH on purification fold and yield in back extraction were also evaluated.
RESULTS: β-mannanase from Proteus vulgaris was purified using aqueous impregnated resin system with a purification fold of 10.7 and yield of 33.7% at 30% (w/w) PEG 4000 (g/mol) impregnated resins, 20% (w/w) of potassium phosphate at pH 7 with 4% (w/w) of NaCl. The back-extraction solution was 20% (w/w) potassium phosphate at pH 7.
CONCLUSIONS: In conclusion, aqueous impregnated resin system offers the possibility to tailor a selective and customized purification system for β-mannanase.

KEY WORDS: Beta-mannosidase; Proteus vulgaris; Polymers