Kinetic characterization of Aspergillus niger chitinase CfcI using a HPAEC-PAD method for native chitin oligosaccharides

Munster, Jolanda M. van; Sanders, Peter; Kate, Geralt A. ten; Dijkhuizen, Lubbert; Maarel, Marc J. E. C. van der

doi:10.1016/j.carres.2015.01.014

Cited by 16 publications

(13 citation statements)

References 34 publications

(36 reference statements)

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“…Using these parameters, symmetrical peaks were obtained for all standards. The COS with the highest DP eluted first, followed by COS with a lower DP, corresponding to results previously described for (GlcNAc) 1–6 with the exception of (GlcN) 1 [37,38] (Figure 1). These elution patterns differ from those obtained for normal homogenous oligosaccharides in ion chromatography, where retention times tend to increase with increasing DP.…”

Section: Resultssupporting

confidence: 86%

“…Recently, Agger reported using HPAEC-PAD to separate COS on theCarboPac-PA1 column, and the monomer peak either partially or completely overlapped with the (GlcN) 5 [38]. On the CarboPac-PA1 column, (GlcNAc) 1 eluted between (GlcNAc) 2 and (GlcNAc) 3 , closer to the latter, when pure water was the mobile phase [37]. Using 5, 15 and 25 mM KOH solutions as the mobile phase, (GlcNAc) 1 always eluted first and before (GlcNAc) 2–6 [38].…”

Section: Resultsmentioning

confidence: 99%

“…High performance anion exchange chromatography (HPAEC) combined with pulsed amperometric detection (PAD) is a powerful, highly sensitive, and efficient system for separating underivatized carbohydrates [34]. Use of HPAEC-PAD for separating COS has previously been reported, but for the most part, they were used only for separating and analyzing (GlcNAc) n [35,36,37,38]. Recently, van Munster reported a quantitative HPAEC-PAD method that allows fast separation of (GlcNAc) 1–6 at the detection limits of 1–3 pmol and a linear range of 5–250 pmol [37].…”

Section: Introductionmentioning

confidence: 99%

“…Use of HPAEC-PAD for separating COS has previously been reported, but for the most part, they were used only for separating and analyzing (GlcNAc) n [35,36,37,38]. Recently, van Munster reported a quantitative HPAEC-PAD method that allows fast separation of (GlcNAc) 1–6 at the detection limits of 1–3 pmol and a linear range of 5–250 pmol [37]. Agger developed a fast and reliable quantitative HPAEC-PAD method for (GlcN) 1 , and the HPAEC elution patterns of (GlcNAc) 1–6 , and of (GlcN) 1,4–6 were investigated [38].…”

Section: Introductionmentioning

confidence: 99%

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Validated HPAEC-PAD Method for the Determination of Fully Deacetylated Chitooligosaccharides

Cao

Wu²,

et al. 2016

IJMS

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An efficient and sensitive analytical method based on high-performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD) was established for the simultaneous separation and determination of glucosamine (GlcN)1 and chitooligosaccharides (COS) ranging from (GlcN)2 to (GlcN)6 without prior derivatization. Detection limits were 0.003 to 0.016 mg/L (corresponding to 0.4–0.6 pmol), and the linear range was 0.2 to 10 mg/L. The optimized analysis was carried out on a CarboPac-PA100 analytical column (4 × 250 mm) using isocratic elution with 0.2 M aqueous sodium hydroxide-water mixture (10:90, v/v) as the mobile phase at a 0.4 mL/min flow rate. Regression equations revealed a good linear relationship (R2 = 0.9979–0.9995, n = 7) within the test ranges. Quality parameters, including precision and accuracy, were fully validated and found to be satisfactory. The fully validated HPAEC-PAD method was readily applied for the quantification of (GlcN)1–6 in a commercial COS technical concentrate. The established method was also used to monitor the acid hydrolysis of a COS technical concentrate to ensure optimization of reaction conditions and minimization of (GlcN)1 degradation.

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Section: Resultssupporting

confidence: 86%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Validated HPAEC-PAD Method for the Determination of Fully Deacetylated Chitooligosaccharides

Cao

Wu²,

et al. 2016

IJMS

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“…A wide range of microrganisms including bacteria, fungi, actinomycetes and yeasts can produce chitinase enzyme [1][2][3][4]. Chitinases are hydrolytic enzymes that cause degradation of β-1-4-glucoside bond of chitin to N-acetyl Dglycosamine [5].…”

Section: Introductionmentioning

confidence: 99%

Mode of Parasitism of the Antagonist Trichoderma Viride towards the Phytopathogen Alternaria Solani

Sharaf

El-Deeb

2018

Journal of Asian Scientific Research

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In a previous work, the optimized crude chitinase of Trichoderma viride exhibited strong inhibitory activity against food spoilage moulds (phytopathogens) Alternaria solani, Botrytis cinerea, Penicillium oxalicum and P. italicum, with the first species being the most affected. Currently, on testing the antagonistic activity of T. viride against the previous phytopathogens, the antagonist exhibited a strong activity particularly towards A. solani. The antagonist displayed more than one mechanism of parasitism on A. solani including penetration of the pathogen hyphae, abnormal pathogen hyphae swelling, lack of conidia formation, conidia malformation as well as degradation of the pathogen cell wall chitin through secretion of chitinase enzyme which is a major cause of antagonism. Fortunately, the present chitinase secreted by Trichoderma viride is found to be a constitutive enzyme. The previously optimized enzyme showed also a powerful insecticidal activity against home insects cockroach, spider and ant where the exoskeleton was softened and hydrolyzed, followed by rapid killing of the insects within 12 minutes. Contribution/ Originality: This study is one of very few studies which have investigated the in vitro mode of parasitism of Trichoderma viride on the food spoilage mold A. solani which exhibited more than one mechanism including hyphae penetration, abnormal hyphae swelling, conidia malformation and lysis of the cell wall chitin.

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Quantitative Kinetic Characterization of Glycoside Hydrolases Using High-Performance Anion-Exchange Chromatography (HPAEC)

McGregor

Arnal

Brumer

2017

Methods in Molecular Biology

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High-performance anion-exchange chromatography coupled to pulsed amperometric detection (HPAEC-PAD) is a powerful analytical technique enabling the high-resolution separation and sensitive quantification of oligosaccharides. Here, we describe a general method for the determination of glycoside hydrolase kinetics that harnesses the intrinsic power of HPAEC-PAD to simultaneously monitor the release of multiple products under conditions of low substrate conversion. Thus, the ability to track product release under initial-rate conditions with substrate concentrations as low as 5 μM enables the determination of Michaelis-Menten kinetics for glycosidase activities, including hydrolysis and transglycosylation. This technique may also be readily extended to other carbohydrate-active enzymes (CAZymes), including polysaccharide lyases, and glycosyl transferases.

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Kinetic characterization of Aspergillus niger chitinase CfcI using a HPAEC-PAD method for native chitin oligosaccharides

Abstract: Citation for published version (APA): van Munster, J. (2014). Carbohydrate-active enzymes that modify the cell wall of Aspergillus niger: Biochemical properties and physiological functions during autolysis and differentiation [S.n.]

Cited by 16 publications

References 34 publications

Validated HPAEC-PAD Method for the Determination of Fully Deacetylated Chitooligosaccharides

Validated HPAEC-PAD Method for the Determination of Fully Deacetylated Chitooligosaccharides

Mode of Parasitism of the Antagonist Trichoderma Viride towards the Phytopathogen Alternaria Solani

Quantitative Kinetic Characterization of Glycoside Hydrolases Using High-Performance Anion-Exchange Chromatography (HPAEC)

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