Exploring combi-lipase catalysis of Eversa® Transform and Candida rugosa lipases for enzymatic hydrolysis of crude palm oil: Effects on fatty acid yield and retention of carotene and tocols

Several lipases have been used to catalyze the hydrolysis of crude palm oil (CPO) to concentrate palm phytonutrients. Previously, the retention of phytonutrients during hydrolysis has been improved with a small addition of ethylenediaminetetraacetic acid. However, the production of free fatty acids (FFA) via enzymatic hydrolysis has yet to be optimised. Thus, for the first time, this study explored the combined use of AY 30SD (from Candida rugosa) and Eversa® Transform 2.0, ET 2.0 (from Aspergillus oryzae) to investigate the effect of reaction time and temperature on the lipases' performance towards the production of FFA during the alcohol-free process of producing phytonutrients concentrates. The results revealed that the application of combi-lipases at 45 °C yielded the highest final FFA content due to ET 2.0's affinity towards longer fatty acid chain lengths and AY 30SD's effectiveness in hydrolysing TAG molecules. With total enzyme concentration of 0.2 wt%, incorporating combi-lipase AY 30SD and ET 2.0 produced >95 wt% of FFA. Furthermore, a basic economic analysis validated that the combination of 0.15 wt% AY 30SD and 0.05 wt% ET 2.0 reduced the reaction time by seven times (completed within 24 h), potentially leading to a substantial increase in profit without sacrificing the phytonutrients retention.