Enzymatic reactions and synthesis ofn‐butyl caproate: esterification, transesterification and aminolysis using a recombinant lipase fromGeobacillus thermoleovoransCCR11

Abstract

The recombinant lipase LipMatCCR11 from the thermophilic strain Geobacillus thermoleovorans CCR11 was applied in the synthesis of n-butyl caproate via transesterification in hexane and xylene. The short chain flavour ester was obtained by alcoholysis from ethyl caproate and n-butyl alcohol and acidolysis from n-butyl butyrate and caproic acid. This enzyme was also used in the condensation reaction from caproic acid and n-butanol. The conversion percentages at equilibrium (Xe) were similar to those obtained with Candida antarctica lipase fraction B (CAL-B) in the same reaction conditions, while lower conversion velocities (k) were attained. LipMatCCR11 reached high conversion percentages in either hexane or xylene as organic media (> 63%); the enzyme was also able to catalyze the aminolysis reaction of ethyl caproate with benzyl amine in hexane obtaining a conversion percentage > 62%.

Polypropylene as a selective support for the immobilization of lipolytic enzymes: hyper‐activation, purification and biotechnological applications

“…34 The use of immobilized enzymes plays a key role in the value chain of biotechnological processes involved in the production of many pharmaceutical products, 35 commodities, 36 in bioremediation, 37,38 biofuels production, 39 food industry, fragrance and flavor compounds, and many other applications. 32,40 Various methods for the immobilization of enzymes exist that comprise binding to a support, entrapment in a carrier, and cross-linking of enzymes. 34,41 The most prominent immobilization methods are adsorption, encapsulation, covalent bonding, and cross-linking.…”

Production and Characterization of Cross-Linked Aggregates of Geobacillus thermoleovorans CCR11 Thermoalkaliphilic Recombinant Lipase

“…3.1.1.3) are ubiquitous enzymes, as they have been found to occur in most living organisms; in vivo, they catalyze the hydrolysis of triacylglycerides to glycerol Molecules 2021, 26, 7569. https://doi.org/10.3390/molecules26247569 https://www.mdpi.com/journal/molecules and fatty acids [1]. In non-aqueous media, lipases can synthesize a wide range of esters or compounds with analogous bonding as amides; due to these features, lipases are widely applied in many industrial areas, such as the food industry, the production of biopharmaceuticals, biofuels, biopolymers, and detergents [2][3][4][5].…”

“…A 43 kDa enzyme, Lip-MatCCR11 has 11 lysine residues in its structure, none of which is directly involved with the active site or the domains of interaction with Ca +2 and Zn +2 , two of the Lys residues located in the lid region. In a previous study, LipMatCCR11 was used in lyophilized form for the synthesis of flavors and fragrance esters with hexane as a reaction medium [3]. Later, using response surface methodology, conditions for maximum expression were investigated and a 1300-fold increase in lipolytic activity was achieved; this enzyme was also immobilized in Accurel EP-100 and used to synthetize aromatic esters [34].…”

Improved expression and immobilization of Geobacillus thermoleovorans CCR11 thermostable recombinant lipase

“…The recombinant protein LipMatCCR11 was the product of the expression system that contained the lipase structural gene minus the signal peptide, and exhibited a lipolytic activity (2.0 × 10 5 U/mg) that was 40‐fold higher than the one produced by the native strain (5,500 U/mg); this recombinant lipase presented an optimal activity under a wide range of temperature (30–60 °C) and pH values (7–10) . In addition, when LipMatCCR11 was used in lyophilized form for the synthesis of flavors and fragrance esters in reactions of acidolysis, alcoholysis, and aminolysis with hexane as a reaction medium, it showed conversion percentages similar to those obtained with Candida antarctica lipase fraction B (CAL‐B) proving to be a new and promising biocatalyst .…”