Optimized butyl butyrate synthesis catalyzed by Thermomyces lanuginosus lipase

Abstract

Butyl butyrate is an ester present in pineapple flavor, which is very important for the food and beverages industries. In this work, the optimization of the reaction of butyl butyrate synthesis catalyzed by the immobilized lipase Lipozyme TL-IM was performed. n-Hexane was selected as the most appropriate solvent. Other reaction parameters such as temperature, substrate molar ratio, biocatalyst content and added water, and their responses measured as yield, were evaluated using a fractional factorial design, followed by a central composite design (CCD) and response surface methodology. In the fractional design 2(4-1) , the four variables were tested and temperature and biocatalyst content were statistically significant and then used for optimization on CCD. The optimal conditions for butyl butyrate synthesis were found to be 48°C; substrate molar ratio 3:1 (butanol:butyric acid); biocatalyst content of 40% of acid mass. Under these conditions, over 90% of yield was obtained in 2 h. Enzyme reuse was tested by washing the biocatalyst with n-hexane or by direct reuse. The direct reuse produced a rapid decrease on enzyme activity, while washing with n-hexane allowed reusing the enzyme for five reactions cycles keeping approximately 85% of its activity.

In Situ Biocatalytic Synthesis of Butyl Butyrate in Diesel and Engine Evaluations

“…In comparison, with the same enzyme, a productivity of 115 mmol L −1 h −1 and 800 mmol L −1 h −1 was reported for the synthesis of butyl acetate in n ‐hexane at an acid concentration of 0.3 m and 2.0 m, respectively, where ultrasound was used to obtain the higher value . For the synthesis of BB by using Thermomyces lanuginosus lipase, a conversion yield of about 90 % was reported after 2 h reaction time at an acid concentration of 0.3 m, which would be equivalent to a productivity of about 135 mmol L −1 h −1 . The productivity was reduced by almost 50 % at 22 °C compared with 37 °C and 60 °C.…”

“…The complex nature of diesel makes it hard to speculate about why the initial rate is higher in hexane but it has been found that longer hydrocarbons negatively influence the activity of lipases. For example, the initial reaction rates of the synthesis of BB using immobilized Thermomyces lanuginosus lipase was 41.7, 48.2, 33.2 mmol h −1 L −1 in n ‐hexane, n ‐heptane, and isooctane, respectively . In addition, there are other compounds and additives in the diesel fuel, which may have negative impacts on the enzyme activity.…”

“…However, the strong incitement for the production of fuels and chemicals through biotechnological means has stimulated the study of lipase‐catalyzed esterification of short chain alcohols and short chain carboxylic acids . BB for example can be enzymatically synthesized from 1‐butanol and butyric acid, both of which can be produced by fermentation technologies using lignocellulosic biomass . In contrast to chemical esterification, enzymatic esterification is highly specific and occurs under mild conditions.…”

High Reusability of Lipase Immobilized on Eggshells for Butyl Butyrate Synthesis under Optimum Condition

“…(Santos and de Castro, 2006). The optimisation of butyl butyrate synthesis catalysed using Thermomyces lanuginosus lipase (TLL) in n-hexane by Martins et al (2013b) produced 90% of the esterification yield and was able to retain 85% of its activity after being used for five times. Martins et al (2013a) reported that the synthesis of butyl butyrate had improved by 2 folds with 3 folds of enzyme activity using the immobilised TLL on the styrene-divinylbenzene beads compared to the commercial Lipozyme TL-IM.…”

Characterization of a new sn‐1,3‐regioselective triacylglycerol lipase from Malbranchea cinnamomea

“…Lipases (EC 3.1.1.3) are enzymes that hydrolyze fatty acid ester bonds in aqueous medium and synthesize them in environment of low water content or in organic solvent medium. Although lipases are widely distributed in animals, plants, and microorganisms, the majority of lipases used for biotechnological purposes are from the proteobacteria ( Burkholderia cepacia , Pseudomonas fluorescens , and Chromobacterium viscosum ), the ascomycete fungi ( Candida antarctica , C. rugosa , Thermomyces lanuginosus , and Aspergillus niger ), and the zygomycete fungi ( Rhizomucor miehei , Rhizopus oryzae , and R. niveus ). The lipase‐catalyzed reactions include hydrolysis, esterification, alcoholysis, acidolysis, interesterification, and aminolysis .…”