Purification and properties of three esterases from Brevibacterium sp. R312

Abstract

C. LAMBRECHTS, J. ESCUDERO AND P. GALZY. 1995. The esterases of Brevibacterium sp. R312 were found to have an intracellular location. Electrophoresis of lysed cell supernatant fluids revealed seven bands of esterase activity in the presence of α‐naphthyl acetate. Eight esterases were separated by anion exchange chromatography. The three main esterases (esterase 4b, 2 and 4a) of Brevibacterium sp. R312 were purified. The molar masses, the pH optima, the temperature optima and heat stabilities were determined. Esterase 2 differed from the two others in sensitivity to inhibitors. Esterase 4b differed from esterases 2 and 4a in its substrate specificity. This enzyme hydrolyses aliphatic and nitrophenyl esters. The spectrum of activity of the two other esterases is narrower. They hydrolysed only naphthyl esters and, in the case of esterase 2, tributyrate and ethyl butyrate.

A new recombinant phosphotriesterase homology protein from Geobacillus caldoxylosilyticus TK4: An extremely thermo- and pH-stable esterase

“…It was reported that Bacillus licheniformis esterase and Brevibacterium sp. R312 esterase 4a had maximum activity at pH 8.0 with p-nitrophenyl caproate and pNPA, respectively [34,35]. buffers.…”

Isolation and characterization of an intracellular esterase fromLactobacillus caseisubsp.caseiIFPL731

“…Lipolysis occurs to a small degree in most cheese varieties and is only especially marked in mould‐ripened varieties, due mainly to very active extracellular lipases produced by Penicillium roqueforti or P. camemberti , and in hard Italian cheeses manufactured with rennet paste containing pregastric lipase and esterases (Fox & Stepaniak 1993). Lactic acid bacteria are considered to be weakly lipolytic compared with the activities of psychrotrophs, micrococci or brevibacteria (Kalogridou‐Vassiliadou 1984; Bhowmik & Marth 1990; Lambrechts et al . 1995), and little is known about their contribution to cheese lipolysis and flavour development.…”

Identification of carboxylesterase activities of commercial triacylglycerol hydrolase (lipase) preparations

“…To determine whether these bands contained catalytic activity of potential relevance to food industry applications and applied enzymology, we subjected them to electrophoretic separation and zymography with alpha-naphthyl acetate, a substrate which has proven useful for the detection of hydrolase activities [15,22,31]. This method was very sensitive, allowing the detection of proteins that were either recalcitrant to silver staining, or present in amounts below the detection limits of silver stain.…”

“…The presence of multiple carboxylesterases in a cheese ripening microorganism, Brevibacterium sp. R312 [22], and Aspergillus niger CLP [13] would exert the same effect.…”

“…Characterization of the selectivity of a C. rugosa CLP has been accomplished using ANA with SDS-PAGE and isoelectric focusing (IEF) separation techniques [21]. ANA has been proven useful in visualizing esterase activity of bacteria beneficial in cheese flavor development [22]. In the present study, ANA zymography was used to positively identify active carboxylesterases (CE) in CLP and to differentiate them from lipases by comparing CE zymograms to earlier work characterizing lipase activities in CLP [2].…”