Combined Effect of Protease, Hemicellulase and Pectinase on the Quality of Hemp Seed Oil (<i>Cannabis sativa</i> L.) Obtained by Aqueous Enzymatic Extraction as an Eco-friendly Method

Abstract

nowadays is Δ9-tetrahydrocannabinol (THC) , present in high proportions in drug hemp but only in trace amounts in fiber hemp.Hemp seeds are well-regarded for their nutritional quality, known for their richness in nutritive elements such as lipids, fiber, protein, vitamins, and minerals. Generally, hemp seeds contain 25-35％ lipids with an excellent balance of fatty acids, 20-25％ proteins with a composition rich in essential amino acids, and 20-30％ carbohydrates, including a substantial amount of dietary fiber 3−5) . Hemp is currently the focus of numerous scientific studies, primari-Abstract: The objective of this research was to evaluate the efficiency of aqueous enzymatic extraction (AEE) to obtain oil from hemp seeds (Cannabis sativa L.) grown in northern Morocco. Optimisation of AEE extraction parameters, including pH, enzyme concentration (hemicellulase, protease and pectinase), temperature and incubation time, to maximize oil yield was achieved using response surface methodology with a central composite design. For comparison, the solvent extraction (Soxhlet) (SE) method was also used. Optimized hydrolysis conditions involved incubation for 4 hours at 60°C with a pH of 6.5, using a multi-enzyme preparation comprising protease, hemicellulase and pectinase at concentrations of 55, 202.5 and 234 U/mg, respectively. Referring to the conventional Soxhlet extraction (SE), Aqueous Enzymatic Extraction (AEE) achieved a 30.65% oil recovery rate under the optimized parameters mentioned above. The use of enzymes produced an oil that was more stable against oxidation than the solvent-extracted oil, with a peroxide value (PV) of 19.54 and 47.87 meq O 2 /kg, respectively. Furthermore, HPLC-DAD analysis of tocopherol content indicated a higher total tocopherol content (547.2 mg/kg) in Aqueous Enzymatic Extraction (AEE) compared to Soxhlet Extraction (SE) (513.51 mg/kg), with γ-tocopherol being the predominant form. No significant differences in fatty acid composition were observed between the two extraction methods with linoleic acid and alpha-linolenic acid being the predominant constituents.