Abstract
The biological activity of apple pectin extracted conventionally or enzymatically using endo-xylanase and endo-cellulase, was tested in vitro. The analyses were performerd in tetraplicates and the statistical significance of the differences were assessed using ANOVA, Tukey post hoc and LSD (the least significant difference) tests. Multivariate regression analysis was applied to determine the structural components that have a crucial importance for antioxidant and antitumor properties of pectins. The pectins extracted by enzymes contained up to four times more ferulic acid and showed twice as great ability to neutralize free radicals and Fe(III) reduction. The antiradical potential positively correlated with phenols, fucose and rhamnose content. In the assays performed on HT-29 human adenocarcinoma and B16F10 melanoma cell cultures, the “green” pectins, contrary to acid isolated ones, exhibited remarkable anti-neoplastic potential while being nontoxic to nontransformed L929 cell line. The pectins in the dose of 1 mg/mL were capable of inhibiting adhesion (max 23.1%), proliferation (max 40.4%), invasion (max 76.9%) and anchorage-independent growth (max 90%) of HT-29 cells (significance level p < 0.001). These pectin preparations were slightly less active towards B16F10 cells. The enzyme-isolated apple pectins may be useful as a functional food additive and an ingredient of the ointment formulas for post-surgical melanoma treatment.
Highlights
Pectins are complex plant polysaccharides and the diversity of their structures depends largely on their origin
The total phenolic content of compounds in apple pomace depends on multiple factors, and varies considerably in literature
Poland; agarose, crystal violet and Coomassie Brilliant Blue G250 were purchased from Merc KGaH, Germany; RPMI 1640 medium and fetal bovine serum (FBS) were from Gibco; glutamine, streptomycin, penicillin, amphotericin B, bovine serum albumin (BSA) and Ca/Mg-containing phosphate buffered saline (PBS) were from Pan Biotech, Germany; sodium pyruvate was from Corning, USA; BD BioCoat Matrigel Invasion Chambers were purchased from BD Bioscience, USA
Summary
Pectins are complex plant polysaccharides and the diversity of their structures depends largely on their origin. Commercial pectins are usually extracted from citrus peel and apple pomace by hot acid (HCl, H2 SO4 and HNO3 ) method at pH 1–3 for 1–12 h at 70–100 ◦ C [2]. The yield of such a process is usually satisfactory, but acid extraction is energy-consuming and increases the risk of accelerated corrosion of production line elements, as well as generates large amounts of acidic industrial waste [3]. Some reports revealed that pectins, highly methylated, decrease the cholesterol, triglyceride, phospholipid and free fatty acid levels in blood plasma and tissues [6]. The in vitro and in vivo studies demonstrated immunomodulatory activity of pectins, which was significant for highly branched polymers [10]
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