Nitrogen sources on TPOMW valorization through solid state fermentation performed by Yarrowia lipolytica

This manuscript reports the valorization of two-phase olive mill waste (TPOMW) as raw material and carbon source for solid state fermentation using Yarrowia lipolytica as biocatalyst. Due to its chemical characteristics, a combination of different raw materials (TPOMW and wheat bran, WB) was evaluated and two distinct nitrogen sources were applied as supplementation for lipase production. A TPOMW/WB ratio of 1:1 and supplementation with ammonium sulfate was chosen as the best condition. The productivity in 24 h reached 7.8 U/gh and, after four days of process, only decreased about 35%. Process pH ranged from 5.5-5.9, remaining in an acid range. Thus, the successful use of TPOMW, a watery solid by-product with high content of lipids, as raw material for Yarrowia lipolytica growth and lipase production provided an environmental friendly alternative to valorize such waste.The authors kindly acknowledge the financial aid and research scholarships given by CAPES. Maria Alice Zarur Coelho thanks CNPq (Proc. 308890/ 2013-2)

Optimization of lipase entrapment in alginate gel bead for palm olein hydrolysis

Lipase from Pseudomonas sp. was entrapped by drop-wise addition of an aqueous mixture of alginate and the biocatalyst to hardening solution of CaCl2 for the purpose of palm olein hydrolysis. Effects ofimmobilization conditions including alginate concentration, CaCl2 concentration, enzyme concentration and bead size on immobilized yield, immobilized lipase activity and recovery of activity (specific activity ratio ofentrapped lipase to free lipase) were investigated. An increase in alginate concentration raised immobilized yield, but decreased immobilized lipase activity and recovery of activity. CaCl2 concentration in the testedrange of 50-200 mM had slight effects on immobilized yield, immobilized lipase activity and recovery of activity. In contrast to immobilized lipase activity, immobilized yield and recovery of activity decreased withincreasing enzyme concentration. With increasing bead size, immobilized lipase activity and recovery of activity decreased due to mass transfer resistance whereas immobilized yield was unchanged. The optimumcondition for lipase entrapment in alginate gel bead was alginate concentration at 2% (w/v), CaCl2 concentration at 100 mM, enzyme concentration at 30 U/ml and bead size at 2 mm. Under this entrapmentcondition, 8.11 U/ml of immobilized lipase was obtained with 95.2% of immobilized yield and 22.2% of recovery of activity

Metagenomic insights into bioaugmentation and biovalorization of oily industrial wastes by lipolytic oleaginous yeast Yarrowia lipolytica during successive batch fermentation

The lipolytic oleaginous yeast Yarrowia lipolytica was used in the bioaugmentation and biovalorization of oily industrial wastes during successive-batch fermentation. Five cycles of nonsterile successive batch fermentation with 70% medium replacement achieved the highest oil removal of 68.1 \ub1 5.60% and produced biomass and lipid yields of 0.213 \ub1 0.07 g/g-COD and 146.2 \ub1 46.5 mg/g-COD, respectively. The cell-bound lipase activity observed in the system was 170.74 \ub1 32 U/L. The auto-flocculation efficiency of the biomass was >90% within 60 Min. The microbial community changes between Y. lipolytica and indigenous microorganisms were monitored by metagenomic next-generation sequencing of internal transcribed spacer rDNA regions for yeasts and 16S rRNA gene for bacteria. Ylipolytica lipolytica was retained in the consortium together with other indigenous strains until the fifth cycle. Other minor oleaginous yeasts such as Kodamaea ohmeri and Candida tropicalis as well as polyhydroxyalkanoate-accumulating bacteria were found and are likely to have participated in lipid production. This study has shown the robustness of Y. lipolytica in nonsterile successive batch fermentation and its use could contribute greatly to the practical valorization of industrial wastes for lipids and lipases