Characterisation and purification of a cinnamate esterase fromAspergillus niger industrial pectinase preparation

Barbe, Christine; Dubourdieu, Denis

doi:10.1002/(sici)1097-0010(199812)78:4<471::aid-jsfa141>3.0.co;2-0

Cited by 27 publications

(10 citation statements)

References 11 publications

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“…The pectinase strength is 3600 MOE units/ml (MOE: Most Einheit = Juice Must Unit; a measure of the reduction in the viscosity of apple juice); the pH and temperature optima of the main activities are 3.5-4 and 45-55°C, respectively. The preparation also harbours cinnamate esterase activity (Barbe and Dubourdieu 1998).…”

Section: Chemicals and Reagentssupporting

confidence: 69%

“…These observations corroborated that p-coumaric acid may be catalytically released from acylated anthocyanins in grape skins, presumably as a result of (9) cinnamate esterase side activity present in the Aspergillus niger pectolytic enzyme preparation (Barbe and Dubourdieu 1998). The data also provided a first indication that p-coumaric acid was apparently converted to caffeic acid in the reaction system by o-hydroxylation catalytic activity.…”

Section: Resultsmentioning

confidence: 99%

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Grape skins (Vitis vinifera L.) catalyze the in vitro enzymatic hydroxylation of p-coumaric acid to caffeic acid

Arnous

Meyer

2009

Biotechnol Lett

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The ability of grape skins to catalyze in vitro conversion of p-coumaric acid to the more potent antioxidant caffeic acid was studied. Addition of different concentrations of p-coumaric to red grape skins (Cabernet Sauvignon) resulted in formation of caffeic acid. This caffeic acid formation (Y) correlated positively and linearly to p-coumaric acid consumption (X): Y = 0.5 X + 9.5; R (2) = 0.96, P < 0.0001. The kinetics of caffeic acid formation with time in response to initial p-coumaric acid levels and at different grape skin concentrations, indicated that the grape skins harboured an o-hydroxylation activity, proposedly a monophenol- or a flavonoid 3'-monooxygenase activity (EC 1.14.18.1 or EC 1.14.13.21). The K (m) of this crude o-hydroxylation activity in the red grape skin was 0.5 mM with p-coumaric acid.

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Section: Chemicals and Reagentssupporting

confidence: 69%

Section: Resultsmentioning

confidence: 99%

Grape skins (Vitis vinifera L.) catalyze the in vitro enzymatic hydroxylation of p-coumaric acid to caffeic acid

Arnous

Meyer

2009

Biotechnol Lett

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“…A. niger CIRM BRFM 131 was also used for genomic DNA extraction, and the resulting DNA was used as a template for PCR amplification. A. niger transformants were grown on selective solid minimum medium (without uridine) containing 70 mM NaNO 3 (29). These ORFs were translated into a set of putative proteins.…”

Section: Methodsmentioning

confidence: 98%

Gene Overexpression and Biochemical Characterization of the Biotechnologically Relevant Chlorogenic Acid Hydrolase fromAspergillus niger

Benoit¹,

Asther²,

Bourne

et al. 2007

Appl Environ Microbiol

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The full-length gene that encodes the chlorogenic acid hydrolase from Aspergillus niger CIRM BRFM 131 was cloned by PCR based on the genome of the strain A. niger CBS 513.88. The complete gene consists of 1,715 bp and codes for a deduced protein of 512 amino acids with a molecular mass of 55,264 Da and an acidic pI of 4.6. The gene was successfully cloned and overexpressed in A. niger to yield 1.25 g liter ؊1 , i.e., 330-fold higher than the production of wild-type strain A. niger CIRM BRFM131. The histidine-tagged recombinant ChlE protein was purified to homogeneity via a single chromatography step, and its main biochemical properties were characterized. The molecular size of the protein checked by mass spectroscopy was 74,553 Da, suggesting the presence of glycosylation. ChlE is assembled in a tetrameric form with several acidic isoforms with pIs of around 4.55 and 5.2. Other characteristics, such as optimal pH and temperature, were found to be similar to those determined for the previously characterized chlorogenic acid hydrolase of A. niger CIRM BRFM 131. However, there was a significant temperature stability difference in favor of the recombinant protein. ChlE exhibits a catalytic efficiency of 12.5 ؋ 10 6 M ؊1 s ؊1 toward chlorogenic acid (CGA), and its ability to release caffeic acid from CGA present in agricultural by-products such as apple marc and coffee pulp was clearly demonstrated, confirming the high potential of this enzyme.

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“…Various methods for esterase activity determination have been reported, such as inter alia, titration (Gilham and Lehner 2005;Schmidt and Bornscheuer 2005), high performance liquid chromatography (HPLC) (Faulds and Williamson 1995;O'Neill et al 1996;Barbe and Dubourdieu 1998;Yue et al 2009), and gas chromatography (GC) (Borneman et al 1990). All ascertain enzyme activity through the determination of acid or other hydrolytes after enzyme action.…”

Section: Introductionmentioning

confidence: 99%

Accurately Determining Esterase Activity via the Isosbestic Point of p-Nitrophenol

Peng¹,

Fu²,

Líu³

et al. 2016

BioResources

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Esterase is an important enzyme for ester hydrolysis or synthesis. Its activity, however, has not been accurately ascertained due to a lack of accurate protocols. In this study, the isosbestic point of p-nitrophenol was found and used as the marker for its activity. The methodology avoided decomposition of the substrate, chromophore agents, and pH changes. The esterase activity was determined accurately and rapidly in a complex solution. In this protocol system, organic solvents were used for dissolving substrates, which influenced activity determination to some extent. Among the solvents tested, methanol exerted the least inhibitory influence. The results indicated that this modified method has potential to be applied for esterase activity determination on a large scale and in real time.

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Characterisation and purification of a cinnamate esterase fromAspergillus niger industrial pectinase preparation

Cited by 27 publications

References 11 publications

Grape skins (Vitis vinifera L.) catalyze the in vitro enzymatic hydroxylation of p-coumaric acid to caffeic acid

Grape skins (Vitis vinifera L.) catalyze the in vitro enzymatic hydroxylation of p-coumaric acid to caffeic acid

Gene Overexpression and Biochemical Characterization of the Biotechnologically Relevant Chlorogenic Acid Hydrolase fromAspergillus niger

Accurately Determining Esterase Activity via the Isosbestic Point of p-Nitrophenol

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