Evaluation of Hydrolysis Conditions for Free Fatty Acid Production Using Immobilized Lipases.
(2025) KBTM01 20251Biotechnology (MSc)
Biotechnology (M.Sc.Eng.)
- Abstract
- Enzymatic hydrolysis using immobilized lipases for the production of free fatty acids and glycerol is a promising alternative to the current industrial standard. This study investigates a set of hydrolysis reactions catalysed by four different immobilization techniques to increase the knowledge on two-phase systems. The evaluation of how water purity, buffer salts, and water amount affect the reaction rate, the pH profile, and extent of enzyme leaching was conducted. The four immobilization techniques displayed variations in results from the studied conditions. Two enzyme immobilizations, X1 and X3, exhibited higher FFA yields with immobilization X1 producing the highest FFA yields with the three conditions: 5 g enzyme with 0.05 M... (More)
- Enzymatic hydrolysis using immobilized lipases for the production of free fatty acids and glycerol is a promising alternative to the current industrial standard. This study investigates a set of hydrolysis reactions catalysed by four different immobilization techniques to increase the knowledge on two-phase systems. The evaluation of how water purity, buffer salts, and water amount affect the reaction rate, the pH profile, and extent of enzyme leaching was conducted. The four immobilization techniques displayed variations in results from the studied conditions. Two enzyme immobilizations, X1 and X3, exhibited higher FFA yields with immobilization X1 producing the highest FFA yields with the three conditions: 5 g enzyme with 0.05 M phosphate, tap water and 0.05 M citrate buffer with 87.4%, 85,7%, and 86,2% FFA yields respectively. Immobilization X3 produced the highest FFA yields when using 5 g of enzyme and 0.05 M phosphate buffer or 10 g of enzyme and DI water to produce 83,2% and 83,2% FFA yields respectively. While immobilization X4, exhibited a pattern similar to the free enzyme as both achieve high initial rates, but sharply curb off around 2 hours achieving much lower final reaction yields compared to the other immobilizations. This effect is likely due to the high concentrations of protein leaching seen by enzyme X4. Through the unique results for each immobilization, more in-depth research can be done to determine significance and determine conditions for further optimization. (Less)
- Popular Abstract
- Fats are found in almost everything we consume as they are natural constituents of both plant-based and animal-based foods. They are often used for cooking and baking to add flavour, texture or balance acidity. Fats are primarily made up of triglycerides, or molecules consisting of three fatty acid chains attached to a glycerol backbone. While fatty acids are often seen in our diets, their relevance does not end there as they also have important applications in industrial products. Fatty acids are key ingredients in a range of products, including detergents, lubricants, plastics, emulsifiers and even tires and candles. Free fatty acids are produced from animal-based or plant-based oils through a process called hydrolysis where the... (More)
- Fats are found in almost everything we consume as they are natural constituents of both plant-based and animal-based foods. They are often used for cooking and baking to add flavour, texture or balance acidity. Fats are primarily made up of triglycerides, or molecules consisting of three fatty acid chains attached to a glycerol backbone. While fatty acids are often seen in our diets, their relevance does not end there as they also have important applications in industrial products. Fatty acids are key ingredients in a range of products, including detergents, lubricants, plastics, emulsifiers and even tires and candles. Free fatty acids are produced from animal-based or plant-based oils through a process called hydrolysis where the triglycerides in the oils are split into free fatty acids and glycerol. Traditionally, this is done using high temperatures and pressures, but this method is energy-intensive and has a significant environmental impact.
A more sustainable alternative for the production of free fatty acids is through a type of enzyme called a lipase. Enzymes are found in all living things, and they act as tiny machines to help get work done like digest food, build new cells, and break down waste. A lipase is a type of enzyme that act like scissors to specifically cut the fatty acids from the glycerol backbone and release them as free fatty acids. This process is more sustainable because it can take place at ambient temperatures and pressures, reducing energy use. These enzymes can be immobilized or trapped to a solid surface or in a material. This allows enzymes to be kept active for longer, by easily separating them from the reaction when it is finished and reusing them for multiple reactions.
This project investigated four different enzyme immobilization techniques, to study how they would react to different reaction conditions. The amount of free fatty acids produced, the pH of the reaction, and the amount of enzyme which leaked out of the immobilization were measured for each condition. During this study, there were different reactions to the conditions between the different lipase immobilizations. One lipase (X4) produced free fatty acids at high rate during the first two hours but subsequently slowed down, while two other lipases (X1 and X3) did not produce at such a high rate in the beginning, but their rate did not slow as much after the first two hours. It is interesting to see how the same enzyme can react so differently depending on the type of immobilization used. Additionally, after studying reactions with waters that differed slightly in ion and salt concentrations, patterns emerged between them. The lipases which were in deionized water, or water containing very little ions or salts, seemed to produce fewer free fatty acids over the reaction time compared to the waters with more ions or salts. From this study, questions were answered as to what factors help the lipases make more free fatty acids and that recycling once is possible, further studies must be conducted to optimize the process before it can become an industrial process. (Less)
Please use this url to cite or link to this publication:
http://lup.lub.lu.se/student-papers/record/9198989
- author
- Bucher, Lydia Elisabeth LU
- supervisor
- organization
- course
- KBTM01 20251
- year
- 2025
- type
- H2 - Master's Degree (Two Years)
- subject
- keywords
- Hydrolysis, Lipase, Immobilization, Biotechnology
- language
- English
- id
- 9198989
- date added to LUP
- 2025-07-01 11:38:27
- date last changed
- 2025-07-01 11:38:27
@misc{9198989,
abstract = {{Enzymatic hydrolysis using immobilized lipases for the production of free fatty acids and glycerol is a promising alternative to the current industrial standard. This study investigates a set of hydrolysis reactions catalysed by four different immobilization techniques to increase the knowledge on two-phase systems. The evaluation of how water purity, buffer salts, and water amount affect the reaction rate, the pH profile, and extent of enzyme leaching was conducted. The four immobilization techniques displayed variations in results from the studied conditions. Two enzyme immobilizations, X1 and X3, exhibited higher FFA yields with immobilization X1 producing the highest FFA yields with the three conditions: 5 g enzyme with 0.05 M phosphate, tap water and 0.05 M citrate buffer with 87.4%, 85,7%, and 86,2% FFA yields respectively. Immobilization X3 produced the highest FFA yields when using 5 g of enzyme and 0.05 M phosphate buffer or 10 g of enzyme and DI water to produce 83,2% and 83,2% FFA yields respectively. While immobilization X4, exhibited a pattern similar to the free enzyme as both achieve high initial rates, but sharply curb off around 2 hours achieving much lower final reaction yields compared to the other immobilizations. This effect is likely due to the high concentrations of protein leaching seen by enzyme X4. Through the unique results for each immobilization, more in-depth research can be done to determine significance and determine conditions for further optimization.}},
author = {{Bucher, Lydia Elisabeth}},
language = {{eng}},
note = {{Student Paper}},
title = {{Evaluation of Hydrolysis Conditions for Free Fatty Acid Production Using Immobilized Lipases.}},
year = {{2025}},
}