Integrated strategy for purification of esterase from Aureobasidium pullulans

Abstract

Esterases catalyze the cleavage and formation of ester bonds of a broad range of substrates presenting a widespread spectrum of industrial applications. This work aimed to partially purify and characterize an esterase from Aureobasidium pullulans LABIOTEC 01 produced in a culture medium containing olive mill wastewater. Esterase purification was evaluated using different strategies, namely the enzyme recovery by PEG-salt aqueous twophase systems (ATPSs); and the enzyme precipitation with ammonium sulfate, acetone, and ethanol. The best purification factor (18 ± 2) was obtained when the ATPS composed of 20% (w/w) polyethylene glycol (PEG) 6000 and 5.8% (w/w) potassium phosphate buffer (PPB) pH 8.0 was combined with acetone precipitation. The partially purified enzyme presented an optimum pH of 5.0, although it remained active in the pH range of 4.5 to 7.5 (≥ 50% relative activity). The optimum temperature was found to be 60°C. Furthermore, the addition of salts such as FeCl 3 , CuSO 4 and MnCl 2 promoted an increase in the enzymatic activity (above 100%). The enzyme was found to be stable and showed high activity when exposed to polar solvents such as dimethyl sulfoxide, dimethylformamide, and methanol. The use of an integrated strategy of purification combining simple purification methods such as ATPS and precipitation was herein reported for the first time for esterase. This strategy proved to be an interesting approach to partially purify the esterase produced under submerged fermentation by A. pullulans. Furthermore, the enzyme showed potential to be applied in industrial biocatalytic processes using high temperature and different salts or solvents. Also, the production of esterase using olive mill wastewater as substrate demonstrated to be a suitable alternative to reduce and valorize agro-industrial residues.

Production and characterization of a thermostable extracellular esterase from Aspergillus niger

“…The result of this study was comparable with the results obtained by Wu et al (2016) in which it was revealed that esterase was stable when enzyme was pre-incubated with 30% iso-propanol, metanol, and n-butanol. Lemes et al (2019) purified esterase from Aureobasidium pullulans and studied its stability towards methanol at high concentration (100%). Adigüzel.…”

Biological Approaches for Extraction of Bioactive Compounds From Agro-industrial By-products: A Review

“…Among the advantages related to enzyme's use at the industrial scale is the cost reduction since enzymes acting as catalysts provide process savings compared to conventional strategies (Singh et al, 2016). To further reduce the cost of applying processes using enzymes, agro-industrial waste available in large quantities can be used in the production of enzymes (Lemes et al, 2016a) using simpler purification protocols or coupling techniques to purify the target product (Lemes et al, 2014;Lemes et al, 2019). Another factor that supports the application of enzymes and cost reduction in the process is the possibility of their immobilization, resulting in their recyclable use, allowing their application in defined cycles, and maintaining their selectivity, catalytic activity, and the generation of products in large quantities (Braga et al, 2014;Basso and Serban, 2019).…”

“…The expenses related to the recovery of target compounds are also positively affected, as enzymatic extracts have a lower cost than pure commercial enzymes. Nevertheless, it is necessary to verify if the application requires purified enzymatic extracts since the inclusion of purification steps would result in a more costly process (Lemes et al, 2014;Lemes et al, 2019).…”

A Temporal Evolution Perspective of Lipase Production by Yarrowia lipolytica in Solid-State Fermentation

“…The global market of enzymes is majorly represented by carbohydrolases, proteases, and lipases, and it was estimated at USD 9.9 billion in 2019 and is expected to grow at a compound annual growth rate of 7.1% from 2020 to 2027 [1,2]. Enzymes have gained interest in the industry mainly due to their widespread uses under mild conditions [3], highlighting the use of lipases among the enzymes.…”