Characterization and

Degradation of Pectic
Polysaccharides in Cocoa Pulp
Esther Meersman, Nore Struyf, Clare Kyomugasho, Zahra Jamsazzadeh
Kermani, Jihan Santanina Santiago, Eline Baert, Sami Hemdane, Gino
Vrancken, Kevin J. Verstrepen, Christophe M. Courtin, Marc Hendrickx, and Jan
Steensels

By Derek Hafstad
Production of Chocolate
Why does it matter?

▶ Chocolate production is a 20 billion dollar a year industry in

the US alone

▶ Could lead to selecting or developing new microbial starter

cultures

▶ Fermenting contributes to the flavor of the chocolate

▶ Can make the production process more efficient

Characterization and Degradation of
Pectic Polysaccharides in Cocoa Pulp
Methods
▶ Cocoa pulp was obtained from the Ivory Coast
▶ Only fresh undamaged pods
▶ Frozen at -20 degrees Celsius
▶ Time between opening pods and freezing was less than 3 hrs
▶ Samples were also pasteurized at 105 degrees Celsius for 5 minutes

▶ Isolation of Cell wall polysaccharides

▶ Extracted as an alcohol insoluble resin (AIR)

▶ Separated by:
▶ Water-extractable pectin (WEP)
▶ Chelator-extractable pectin (CEP)
▶ Sodium carbonate extractable pectin (NEP)
▶ Hemicellulose fractions (HF)
Analyzing The Samples
▶ Determination of Galacturonic Acid (GalA)- spectrophotometrically with m-
hydroxydiphenyl method
▶ Determination of the Degree of Methyl Esterification (DM)- Fourier transform
infrared spectroscopy
▶ Determination of Molecular Mass Distribution- high performance size exclusion
chromatography and multiangle laser light scattering
▶ Analysis of Neutral Sugar Content- high performance anion exchange
chromatography with a pulsed amperometric detector
▶ Determination of Protein Content- Dumas protein analysis system
▶ Determination of Reducing End Sugar Residuals- gas chromatography
Results

Composition of Pectin Fragments

WEP and CEP most abundant

36.2% and 35.5% respectively

Small amounts of NEP and HF

Sugar Content
Effects of pH and Temperature on
Viscosity

The effect of temperature and pH examined because it

changes due to the fermentation
Effects of Enzymatic Activity

AFA: α-L-Arabinofuranosidase

EPG: endopolygalacturonase

BX: β-D-xylosidase

XYL: endo-1,4-β-xylanase
Endopolygalacturonase Activity of Yeast

.
 Significance

▶ The study provides a structural analysis of pectin components

▶ Shows the structural complexity of cocoa pulp
▶ High amounts of weakly and ionically bound pectic polysaccharides and small
amounts of strongly ester bound pectic polymers and hemicellulosic polymers
▶ Shows increase temperature and endopolygalacturonases may contribute to
pulp liquefaction
▶ Shows endopolygalacturonases contributes to pulp breakdown by cleaving high
molecular weight pectin molecules
My thoughts

▶ Pro:
▶ Very thorough in the methods

▶ Pro/Con:
▶ More focused towards biochemists

▶ Con:
▶ There are times where they are not very clear
Questions?
References
▶ Meersman, Esther, et al. “Characterization and Degradation of Pectic
Polysaccharides in Cocoa Pulp.” Journal of Agricultural and Food
Chemistry, vol. 65, no. 44, 2017, pp. 9726–9734.,
doi:10.1021/acs.jafc.7b03854.

▶ Chocolate Production,
nikitahuggins.com/Projects/Food%20Systems/Chocolate/production.html.
▶ Davidson, Michael W. “Plant Cell Wall.” Molecular Expressions Cell Biology:
Plant Cell Structure - Cell Wall, Florida State University,
micro.magnet.fsu.edu/cells/plants/cellwall.html.
▶ Index of /Wp-Content/Uploads/2010/06, blog.rococochocolates.com/wp-
content/uploads/2010/06/.