Extraction and characterization of pectins from agricultural byproducts;

conventional chemical versus eco-friendly physical/enzymatic treatments

J ongbin Lim
a
, Bockki Min
a
, Yu J eong Kim
a
, Sanghoon Ko
a
, Choon Gil Kang
b
, and Suyong Lee
a

a
Department of Food Science and Technology, Carbohydrate Bioproduct Research Center, Sejong
University, Seoul 143-747, Republic of Korea (suyonglee@sejong.ac.kr)
b
Research Center, Ottogi corporation, Kyeonggi-Do 431-070, Republic of Korea


ABSTRACT
Pectin from Yuza pomace which is the main waste of fruit juice industry was extracted in an environmentally
friendly way and their structural and physicochemical properties were then compared with those of
chemically-extracted pectin. The combined physical/enzymatic treatments produced pectin with 55%
galaturonic acid content and 7.3% yield which were lower than those of the chemically-extracted pectin. The
pectin sample gained by the physical/enzymatic treatments showed higher degree of esterification (46.30%)
than the chemically-extracted pectin (40.93%) which was also confirmed by FT-IR analysis. When subjected
to steady-shear rheological condition, both pectin solutions were shown to have shear-thinning properties.
Thus, the use of green-labelled pectin can allow the food industry to move toward environmentally friendly
technology and sustainable production.

Keywords: Yuza; Pectin; Eco-friendly; Agricultural by products; Rheology


INTRODUCTION
Yuza (Citrus junos) pomace that accounts for about 10 ~15% of Yuza fruit is generated from Yuza juice
extraction[1] and approximately 1,800 tons of the pomace per year are wasted[2]. However, Yuza pomace
with unique flavor contains a variety of useful components. Especially, pectin has become highly valued as a
soluble dietary fiber. Therefore, various efforts to utilize pectin from Yuza pomace have been made. As a
natural and complex polysaccharide, pectin is found primarily in the cell walls where they play an essential
role in cell to cell adhesion[3]. Also, from a food industrial point of view, pectin has been extensively used in
a number of food products such as ice creams, beverages, jellies, and sources due to its typical gelling,
thickening, and stabilizing properties. Pectin has been industrially obtained in a chemical way with strong
acids such as oxalic[4], hydrochloric[5], and sulfuric acid[6]. Thereby, the use of the chemicals may probably
impair the functionality of pectin and cause environmental problems. Therefore, the eco-friendly treatments
have been taken into account for minimizing the use of detrimental chemicals. For examples, natural
catalysis with hemicellulose, protease[7], and microbial enzymes[8] has been applied. Also, several studies
on thermo-mechanical technology such as ultrasound[9], autoclaving[10], and extrusion[11], have been
reported. However, the previous research focused primarily on the yield and chemical composition of the
extracted pectin. Moreover, combined physical/enzymatic treatments have not been carried out yet for
extracting pectin from agricultural byproducts. The aims of this study were to establish the eco-friendly
extraction process by combined physical/enzymatic treatments and to investigate the chemical, structural, and
rheological properties of the extracted pectin.

MATERIALS & METHODS
Materials
Yuza pomace provided by Hansung Food Co. (J eollanam-do, Korea) was dried in an oven at 80 for 24 h
and ground to pass through a 100 mesh sieve.

Pectin extraction
Extraction of pectin from Yuza pomace was performed in two different ways, which were chemical and
combined physical/enzymatic methods. In case of the conventional chemical method[4], the dried Yuza
pomace powder was treated with 85% ethanol at 70 for 20 min and filtered with miracloth (Merck KGaA,
Darmstadt, Germany). The residue (10 g) was stirred with oxalic acid/ammonium oxalate (0.25%, pH 4.6,
400 mL) at 85 for 1 h. The compound was filtered with miracloth, and the filtrate was mixed with three
volumes of 96% ethanol. After centrifugation at 14,500 x g for 10 min, the precipitation was washed with a
series of 70%/96% ethanol and then oven-dried at 50 for 24 h. The combined physical/enzymatic
treatments were also applied to obtain the eco-friendly extracted pectin. The suspension of Yuza powder in
distilled water (5%, w/v) was filtered with miracloth and the residue was mixed with distilled water (5%,
w/v). After autoclaving at 121 for 5 min, the Viscozyme

L (Novozymes, Bagsvaerd, Denmark) with 1.2

10
-4
fungal -glucanase unit was added into the suspension, which was kept at 40 for 1 h. Finally, the
suspension was dialyzed (MW 8,000 cutoff, Spectrum Laboratories Inc., CA, USA) and freeze-dried.

Chemical and structural analysis
The degree of esterification of the extracted pectins was measured according to the method of Klavons et
al[12]. The content of galacturonic acid was also determined based on the method of Cozzy and Carpita[13].
The structure of the pectins extracted by two different methods was performed by using a FT-IR spectrometer
(Nicolet Instrument Co., Madison, WI, USA).

Rheological measurements
Rheological measurements were made by using a controlled-stress rheometer (AR1500ex, TA Instruments,
USA) with a 40 mm parallel plate. Shear rates (0.1 - 200/s) were applied for steady-shear viscosity at 25
which were then fitted into the Cross equation.

RESULTS & DISCUSSION
Fig. 1 shows the extraction procedures of pectin from Yuza pomace by two different ways.

























Figure 1. Schematic of pectin extraction from Yuza pomace in (a) chemical and (b) eco-friendly ways.

As presented in Table 1, the degree of esterification of the pectins extracted by chemical and combined
physical/enzymatic treatments was determined to be 40.93% and 46.30%, respectively. The previous study
reported that the degree of esterification of Yuza pectin extracted by citrate, tartrate, and HCl was less than
50%, which was in a good agreement with our study. Thus, the pectins extracted by combined
physical/enzymatic treatments could be classified as low methoxyl pectin. Also, the yield of the eco-friendly
extracted pectin was 7.33%, which was quite similar to that of chemical-extracted pectin. It was reported that
the yield of the pectin from Yuza was typically 4 19%, depending on the extraction methods[14].
The galacturonic acid content of the chemically-extracted pectin was 72.33% while that of the pectin
obtained by combined physical/enzymatic method was 55%. It would be probably due to the increased
solubility of non-pectinous polysaccharides by the combined physical/enzymatic treatments[15].

Table 1. Degree of esterification, yield, and galacturonic acid content of Yuza pectin extracted by chemical and
physical/enzymatic treatments.
Characteristics Chemical extraction Physical/enzymatic extraction
Degree of esterification (%) 40.93 46.30
Yield (%) 8.00 7.33
Galcturonic acid content (%) 72.33 55.00

Fig. 2 exhibits the FT-IR spectra of the pectins which were extracted by two different methods. A band at
3350 cm
-1
was observed, which corresponded to hydroxyl groups (OH). In addition, O-CH
3
bond stretching
from methyl esters of galacturonic acids was detected at 2930 cm
-1
. It was interesting to note that two bands
associated with ester carbonyl group and free carbonyl group exhibited at 1740 cm
-1
and 1610 cm
-1
,
respectively.






















Figure 2. FT-IR spectra of Yuza pectin which were extracted by chemical and physical/enzymatic treatments.

The flow behaviors of the pectins extracted by two different methods were investigated under steady-shear
conditions. The apparent viscosity of pectins was increased with an increase in pectin concentration.
Furthermore, the apparent viscosity decreased with increasing shear rates for all samples, showing their
shear-thinning features. Especially, these shear-thinning features became more apparent with increasing
concentrations. The pectin which was extracted by chemical treatment showed Newtonian plateau at low
shear rates and then exhibited shear-thinning features with increasing shear rates. While the pectin obtained
by combined physical/enzymatic treatments appeared shear-thinning features in the range of shear rates
tested in this study which were characterized by the Cross equation as follows;






















Figure 3. Flow behaviour of Yuza pomace pectin extracted by chemical and physical/enzymatic treatments.

CONCLUSION
The combined physical/enzymatic treatments were applied to extract pectin from Yuza pomace in an eco-
friendly way. Hydrothermal treatments with enzymatic assistance produced low methoxyl (46.3%) Yuza
pectins with 55% galacturonic acid. The pectins extracted by the combined physical/enzymatic treatments
showed shearing-thinning properties which were fitted well into the Cross equation. The eco-friendly pectin
would be utilized to control the rheology and texture of food products as much as chemically-extracted pectin
and furthermore provide positive benefits with possible natural green-labeling.

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