SCALING UP β-FRUCTOSIDASE PRODUCTION BY PICHIA PASTORIS BfrA4X ON A COST-EFFECTIVE CANE MOLASSES MEDIUM

Abstract

Introduction:
Molasses is an agro-industrial by-product that could be used as inexpensive carbon source to reduce the culture media cost for enzyme-producing microorganisms.
Objective:
To evaluate four combinations of carbon and nitrogen sources for industrial production of BfrA by Pichia pastoris BfrA4X.
Materials and Methods:
Medium composition was evaluated in an orbital shaker and increased discontinuous cultures. The culture was scaled up to a 42-L fermenter based on a constant volumetric mass transfer coefficient (kLa). Enzyme activity and biomass yield was evaluated.
Results and Discussion:
When increased discontinuous cultures was carried out in a 7.5-L fermenter with MYH medium, an overall BfrA yield of 118 ± 1 U/mL of fermentation culture was obtained, similar to the yield of 120 ± 7 U/mL yield obtained for SYE medium. The culture was expanded to a 42-L fermenter, with MYH medium a higher overall BfrA activity of 167 ± 8 U/mL was obtained.
Conclusions:
Cane molasses is a low-cost feedstock that could be used for the economic scale-up of efficient BfrA production by P. pastoris BfrA4X, based on constant kLa.