Use of grape pomaces to produce biomass of a Komagataella pastoris strain expressing a bovine chymosin activity

D. KINGSTON, G. F. NOVELLI, P. CERRUTTI, M. N. RECUPERO, M. BLASCO Y M.A. GALVAGNO

CRITICAL REVIEWS IN FOOD SCIENCE AND NUTRITION

TAYLOR & FRANCIS INC

Año: 2014 vol. 2 p. 734 - 743

1040-8398

The use of agro-industrial wastesnot only decreases bioprocesses and disposal costs but also contributes to theupgrading of the residues. An active recombinant methanol-inducible bovinechymosin has been expressed in our laboratory in the yeast Komagataella pastoris, andgrape pomaces extracts (GRE) were proposed as a convenient C-energy source forthe biomass production of the genetically engineered strain.Carbon and nitrogen sources, growthfactors and initial pH conditions were selected by classical methodology;thereafter, growth conditions optimization was performed using StatisticalDesigned Experiments (DoEs). In the presence of (g L-1): 67.0monosaccharides (glucose and fructose) from GRE; 5.0 (NH4)2SO4and 10.0 sugar cane molasses (CMz), a yield of 20.0 g L-1 celldry weight (CDW) was obtained aerobically after 60 h incubation at 28ºC and pH 4.0. Applying afed-batch strategy with methanol:sorbitol as the enzyme inducers, a chymosinproduction of 8.53 International-Milk-Clotting-Units (IMCU) per mg protein wasobtained in the supernatant.The results presented show thatthrough a statistical design, a simple, cheap and easy to prepare culturemedium could be developed using two agroindustrial derivatives (GRE and CMz) toobtain a higher value added product.