Preparation of high-quality sunflower seed protein with a new chlorogenic acid hydrolase from Aspergillus niger

ABSTRACT 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 × 106 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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Weitere Charakterisierung einer Chlorogensäure-Hydrolase aus Aspergillus niger / Further Characterization of a Chlorogenic Acid Hydrolase from Aspergillus niger

Zeitschrift für Naturforschung C ◽

10.1515/znc-1980-9-1006 ◽

1980 ◽

Vol 35 (9-10) ◽

pp. 699-701 ◽

Cited By ~ 15

Author(s):

B. Schöbel ◽

W. Pollmann

Keyword(s):

Activation Energy ◽

Molecular Weight ◽

Aspergillus Niger ◽

Chlorogenic Acid ◽

Sodium Dodecyl ◽

Acid Hydrolase ◽

Ester Linkage ◽

Total Molecular Weight ◽

Ph Range

Abstract In addition to our previous paper [1] further characteristics of the chlorogenic acid hydrolase are described. Polyacrylamid gelelectrophoresis revealed only one band for the purified enzyme. Sodium dodecyl-sulfate polyacrylamid gelelectrophoresis showed a molecular weight of 60000, demonstrating four subunits of the enzyme (total molecular weight 240000). The enzyme is stable in a pH-range of 3 .0 -8 .5 and up to a temperature of 55 °C. The temperature coefficient Q10 is 1.5, the activation energy EA is 6.0 kcal/mol. The amino acid analysis and substrate specificity data are given in tables. Essential for the enzyme activity is the C=C double bound neighbouring the ester linkage. The enzyme crystallizes in prisms.

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Purification and characterization of a chlorogenic acid hydrolase from Aspergillus niger catalysing the hydrolysis of chlorogenic acid

Journal of Biotechnology ◽

10.1016/j.jbiotec.2004.07.009 ◽

2005 ◽

Vol 115 (1) ◽

pp. 47-56 ◽

Cited By ~ 31

Author(s):

Michèle Asther ◽

Maria Isabel Estrada Alvarado ◽

Mireille Haon ◽

David Navarro ◽

Marcel Asther ◽

...

Keyword(s):

Aspergillus Niger ◽

Chlorogenic Acid ◽

Acid Hydrolase ◽

Purification And Characterization ◽

Hydrolysis Of

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Effect of free amino acids and peptide hydrolysates from sunflower seed protein on the formation of pyrazines under different heating conditions

RSC Advances ◽

10.1039/d1ra05140g ◽

2021 ◽

Vol 11 (45) ◽

pp. 27772-27781

Author(s):

Furong Wang ◽

Hailiang Shen ◽

Xi Yang ◽

Ting Liu ◽

Yali Yang ◽

...

Keyword(s):

Amino Acids ◽

Maillard Reaction ◽

Free Amino Acids ◽

Free Amino ◽

Sunflower Seed ◽

Seed Protein ◽

Heating Temperature ◽

Reaction Model

Exploring the effect of heating temperature and time on the formation of pyrazines; revealing the potential roles of FAAs and hydrolyzed sunflower seed peptides in the Maillard reaction model.

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CHLOROGENIC ACID CONTENT OF SUNFLOWER SEED FLOUR AS AFFECTED BY SEEDING AND HARVEST DATE

Canadian Journal of Plant Science ◽

10.4141/cjps76-146 ◽

1976 ◽

Vol 56 (4) ◽

pp. 901-905 ◽

Cited By ~ 4

Author(s):

D. G. DORRELL

Keyword(s):

Chlorogenic Acid ◽

Acid Content ◽

Vegetative Growth ◽

Sunflower Seed ◽

Growing Season ◽

Growing Degree Days ◽

Degree Days ◽

Normal Field ◽

Seed Flour ◽

Seeding Date

The effect of seeding date on the chlorogenic acid content of sunflower seed flour was determined by seeding the cultivars Krasnodarets and Peredovik at seven dates, starting on 14 May, over 3 yr. Sequential plantings were made at increments of approximately 70 growing degree days (base = 5.6 C). Plants were harvested at normal field maturity. The time and rate of deposition of chlorogenic acid was determined by harvesting plants at 7-day intervals from 21 to 49 days after flowering. The seeds were dehulled and defatted before determining the chlorogenic acid content of the flour. Chlorogenic acid content declined steadily from an average of 4.22% for the first seeding to 3.30% for the last seeding. About one-half of the total chlorogenic acid was present 21 days after flowering. Deposition continued rapidly for the next 14 days then the level began to stabilize. Delay in seeding tended to shorten the period of vegetative growth and shift the deposition of chlorogenic acid to a cooler portion of the growing season. It is suggested that a combination of these factors caused the reduction in chlorogenic acid content of sunflower flour.

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