Esterase screening using whole cells of Brazilian soil microorganisms

Abstract

Um ensaio enzimático miniaturizado com sondas fluorescentes para triagem rápida de microrganismos produtores de esterases foi implementado e aplicado a 64 linhagens de bactérias de solo. Os melhores resultados foram validados por métodos convencionais e substratos não fluorogênicos (feniletanol acetilado e propanoilado) e confirmaram os resultados obtidos nas triagens rápidas. Os microrganismos que apresentaram atividades enzimáticas mais relevantes (razão enantiomérica E > 30) na hidrólise de ésteres e na obtenção de boas razões enantioméricas foram identificados como pertencentes ao gênero Bacillus. Parte dos microrganismos se encontra preservada em glicerol e/ou liofilizada no IQ/Unicamp e três linhagens serão depositadas na CBMAI do CPQBA/Unicamp como depósitos abertos.A miniaturized enzymatic assay using fluorescent probes to reveal esterase producing microorganisms was optimized and applied to screen 64 soil bacterial strains. The best results were validated using traditional non-fluorogenic assays with acetyl and propanoyl phenylethanol to confirm the miniaturized results. The most active microorganisms belong to the genus Bacillus showing esterase activity and good enantiomeric ratios for the resolution of phenylethanol derivatives (E > 30). Part of the microorganisms are kept in our laboratory in glycerol or freezedried and the best microorganisms will be deposited in the CBMAI/CPQBA/Unicamp culture collection.

Design of Immobilized Enzyme Biocatalysts: Drawbacks and Opportunities

“…[1][2][3][4][5][6][7][8] The enzymes are used industrially, among others, in food products, cellulose and paper, animal feed, textile cosmetics, detergents and the pharmaceutical industry. [9][10][11][12][13][14][15][16] To increase the use of enzymes as industrial biocatalysts, it is necessary to obtain stable preparations of the enzymes with an improved operational stability, 17 their application renders into lower demand for raw materials, chemicals and energy, heir specificity and high selectivity, and they drive into cleaner industrial processes. 18 However, immobilization is one of the important ways for enzymes to become stable.…”

Fungal mediated kinetic resolution of racemic acetates to ( R )-alcohols using Fusarium proliferatum

“…Enzyme mediated kinetic resolution of such alcohols using lipases, esterases, and alcohol dehydrogenases have been reported in the literature. [9][10][11][12][13][14][15][16][17][18][19][20][21][22][23][24][25][26][27][28] For example, resolution of (±)-lavandulol has been achieved previously with various lipases such as Candida antarctica lipase B (CAL B), 9 Hog pancreas lipase, 29 Porcine pancreas lipase, 26 and Yarrowia lipolytica lipases. 30 However, enzyme catalyzed resolution process is associated with several disadvantages such as higher cost, low substrate concentration, instability and cofactor dependency of the enzymes in several occasions.…”

Hydrocarbon-associated substrates reveal promising fungi for poly (ethylene terephthalate) (PET) depolymerization

“…Fungal suspensions (1 mg.mL −1 ) were prepared in borate buffer (pH 7.4). The assays were performed on 96-well flat bottom microtiter plates, as proposed by Mantovani, Oliveira, and Marasaioli [46]; for esterase activity, Probe 1-short-chain ester, namely, 4-[(2-oxo-2H-1-benzopyran-7-yl)oxy]butane-1,2diylacetate, was used, and probe 2-long-chain ester, 4-[(2o x o -2 H -1 -b e n z o p y r a n -7 -y l ) o x y ] b u t a n e -1 , 2diyldioctanoate, for lipase activity (Fig. 1).…”