Immobilization of Moniliella spathulata R25L270 Lipase on Ionic, Hydrophobic and Covalent Supports: Functional Properties and Hydrolysis of Sardine Oil

Abstract

The oleaginous yeast Moniliella spathulata R25L270 was the first yeast able to grow and produce extracellular lipase using Macaúba (Acrocomia aculeate) cake as substrate. The novel lipase was recently identified, and presented promising features for biotechnological applications. The M. spathulata R25L270 lipase efficiently hydrolyzed vegetable and animal oils, and showed selectivity for generating cis-5,8,11,15,17-eicosapentaenoic acid from sardine oil. The enzyme can act in a wide range of temperatures (25–48 °C) and pH (6.5–8.4). The present study deals with the immobilization of M. spathulata R25L270 lipase on hydrophobic, covalent and ionic supports to select the most active biocatalyst capable to obtain omega-3 fatty acids (PUFA) from sardine oil. Nine immobilized agarose derivatives were prepared and biochemically characterized for thermostability, pH stability and catalytic properties (KM and Vmax). Ionic supports improved the enzyme–substrate affinity; however, it was not an effective strategy to increase the M. spathulata R25L270 lipase stability against pH and temperature. Covalent support resulted in a biocatalyst with decreased activity, but high thermostability. The enzyme was most stabilized when immobilized on hydrophobic supports, especially Octyl-Sepharose. Compared with the free enzyme, the half-life of the Octyl-Sepharose derivative at 60 °C increased 10-fold, and lipase stability under acidic conditions was achieved. The Octyl-Sepharose derivative was selected to obtain omega-3 fatty acids from sardine oil, and the maximal enzyme selectivity was achieved at pH 5.0.

Co-Immobilization of Tri-Enzymes for the Conversion of Hydroxymethylfurfural to 2,5-Diformylfuran

“…The temperature is an important parameter for the properties of enzymes [49,50,51,52]. The DFF yield was studied as a function of temperature ranging from 15–45 °C.…”

Immobilization of Enzymes on a Phospholipid Bionically Modified Polysulfone Gradient-Pore Membrane for the Enhanced Performance of Enzymatic Membrane Bioreactors

“…The construction of an in vitro biofriendly microenvironment plays an important role in maintaining the high catalytic activity of the enzyme, and surfactants are often used to form enzyme surfactant micelles that show significantly higher efficiency [ 29 , 30 ]. It has been confirmed that the higher activity is a result of the structure of the surfactant and the interaction between the micelle-bound enzyme and the substrate.…”

“…The main type of interaction between the enzyme and lecithin is electrostatic, because of the large number of positive and negative charges on the surface of the enzyme and lecithin, which can alter the conformation and activity of the enzyme to some extent. In particular, at low concentrations, lecithin, contains a monovalent quaternary ammonium cation, binds to the lipase through weak electrostatic interactions, which alter the conformation of the enzyme and allow it to combine with the substrate more easily [ 30 ]. Nevertheless, as the concentration of lecithin increased, a decay phenomenon emerged in the relative activity of the lipase.…”

Waste Management in the Agri-Food Industry: The Conversion of Eggshells, Spent Coffee Grounds, and Brown Onion Skins into Carriers for Lipase Immobilization

“…Lipases are used in free form as an active ingredient in foods, such as bakery and pasta products, and various types of cheese and dairy products to improve the quality and taste of the final product [24,[26][27][28] or in immobilized form to provide a substitute for human milk [27,28], cocoa butter [29][30][31][32], specific flavors as food additives [33], or for the production of ω-3-and ω-6-fatty acids from fish and other oil species [34,35]. Although lipase accounts for about 10% of all industrial enzymes used in current industrial production, the use of these enzymes is expected to increase due to their versatility [24,[36][37][38][39].…”