Lipase Production

Lipases (triacylglycerol acylhydrolases, EC. are important enzymes due to their various industrial applications. The ability of lipases to catalyze both hydrolytic and synthetic reactions has directed research towards seeking better understanding of their properties as well as their potential applications. Despite the great industrial potential of lipases, the high production cost of lipases limits their industrial applications. Recently, lipases have been produced using microbial sources along with agro-industrial wastes to reduce the production cost especially employing solid-state bio-conversion for the process. Based on that, I was working on Palm kernel cake (PKC) as one of the solid wastes obtained after oil extraction from palm kernel seeds. The nutritive value of PKC directed studies towards utilization of this substrate in order to produce enzymes as it is a rich source of carbohydrates as well as protein and minerals which makes it suitable for microbial fermentation. All pictures provided are the author's copyright and taken during the experimental sessions.

PKC collection at Sime Darby plantation, Kery Island, 2012
Solid-state Fermentation of the palm kernel cake (PKC) for 72 hours.
Crude Lipase obtained after the filtration of the extract.
Lipase assayusing pNPP as a substrate to release pNP
Obtaining solid lipase after freeze drying
Lipase after drying
Hydrolysis of different oils using the produced lipase
The results after titration. See the light pink color for the end point

Ionic liquid-Compatible Cellulase

The pesearch involves bioethanol production with details on the production of cellulase enzyme from Trichoderma reesei (RUTC30) from agricultural residues and lignocellulosic biomass and conducting the synthesis of the ionic liquids from their raw materials. The aim is to find a suitable and compatible ionic liquid that can stabilize the cellulase enzyme to conduct a single-step hydrolysis followed by fermentation of the hydrolysate using yeast.

Cellulase production

Trichoderma reesei ;

5-day culture


Trichoderma reesei (microscopic image); 5-day culture

Solid-state fermentation of rice husk by Trichoderma reesei

Solid-state fermentation of PKC by Trichoderma reesei
Solid-state fermentation of empty fruit bunch (EFB) by Trichoderma reesei
Crude and purified PKC-Cellulase (PKC-Cel)
Micro-filtration and Ultra-filtration of PKC-Cel
Cellulase assay; DNS Method

Ionic Liquid Synthesis

Ionic liquid synthesis by drop-wise addition
Ionic liquid synthesis; reaction completion
Ionic liquid synthesis; evaporation of the solvent
Choline acetate [Cho]OAc Ionic liquid
Confirmation of synthesis using NMR at UTP facilities.

Biomass Pretreatment, Enzymatic Hydrolysis & Fermentation

Determination of structural carbohydrates
Determination of cellulose
Determination of cellulose
Heating EFB with ILs
Addition of PKC-Cel and buffer
Determination of reduction sugar
Saccharomyces cerevisiae5-day culture
Analysis with GC-MS

Deep Eutectic Solvents

Enzyme stability in deep eutectic solvents

Ionic Liquids Applications

Project title: Development of Future Antibiotics: Active Pharmaceutical Ingredient Ionic Liquids (API-ILs) with COSMO-RS Based Prediction for Resistant Microbial strains.

Nanoemulsions (NEs)

Project title: Nanoemulsions Production Using Deep Eutectic Solvents on Fish By-Products