1. Optimization of Solid Fermentation Medium
Composition by Simplex Centroid Mixture Design
Method for Lipase Production using a New
Penicillium sp. and Agroindustrial Subproducts as
Substrates
Camilo Teixeira, Débora Moreira, Paula Speranza, Gabriela Macedo
Food Science Department, School of Food Engineering (FEA), University
of Campinas (UNICAMP)
2. Introduction
Aims to develop clean and efficient processes
for biomass conversion into low-cost and
sustainable biofuels, chemicals and
pharmaceuticals.
3. Biocatalysis: Lipases and its potential applications
• Triglycerides hydrolysis
• Biosynthesis: Esterification, Interesterification, etc.
• Detergents, Biodiesel, Cosmetics, Food processment
Lipase
4. Solid state fermentation (SSF) enzyme production
• Agroindustrial byproducts as Fermentation Medium;
• Lower energy cost than submerged fermentation;
• Considered as a green technology
5. Objectives
• Feedstock selection for SSF lipase production: wheat bran, soybean
meal and cottonseed meal;
• Evaluate the best formulation by the Simplex Centroid Mixture Design
(SCMD)
• Simultaneous optimization of lipase activity and productivity by the
desirability function;
6. Materials
• Soybean meal and cottonseed meal were supplied by Bunge
Brazil (Rondonópolis, Mato Grosso, Brazil)
• Wheat bran was purchased in local Market (Campinas, São
Paulo, Brazil)
• The Penicillium sp. strain was isolated by Dr. Luciana Fleuri
(UNESP – Botucatu, São Paulo, Brazil)
11. Solid State Fermentation
250mL erlenmeyer with cotton-wool plug
1 mL of inoculum solution
10g of different mixtures of WB, SM and CSM
10g water (1:1 mass:water)
Climatic Chamber
96 Hours 30ºC 90% humidity
13. Lipase Hydrolytic Activity (Titrimetric Method)
1 mL filtrate
30 minutes 37ºC pH 7
Olive oil + Arabic Gum + Phosphate buffer pH 7
Stopped with etanol: acetone solution
NaOH 0.05 M
Phenolphthalein 1%
Sample – Control (without enzyme)= [fatty acid]
1 U = 1 µmol oleic acid/minute
14. Experimental Design by Simplex Centroid Mixture
Method
• Determine the best formulation for a mixture of components;
• Wheat Bran, Soybean Meal and Cottonseed Meal as independent
variables
• Lipase Activity (U/mL) and Productivity (U/R$) as response variables;
• Evaluate the best mixture and possible interactions with a positive
synergistic effect for the response variables;
26. Conclusions
• Lipase Activity is directly correlated with feedstock lipid
concentration;
• Cottonseed Meal have the highest lipid concentration, lipase activity
and productivity.
• The mixture of soybean meal, cottonseed meal and wheat bran did
have a small synergistic positive effect
• The desirability function can optimize simultaneously the lipase
activity and the productivity responses.
27. Food Biochemistry Laboratory, Food Science
Dept., School of Food Engineering, UNICAMP
Facebook: Bioquímica-FEA/UNICAMP
