Impact of Cabbage Pectin–Protein Complex on Microbial β‑Glucuronidase Activity

• Published in: Journal of agricultural and food chemistry 2013-03, Vol.61 (12), p.3054-3058
• Main Authors: Borisenkov, Mikhail F, Bakutova, Larisa A, Golovchenko, Victoria V, Vityazev, Fedor V, Patova, Olga A, Ponomareva, Svetlana A, Popov, Sergey V
• Format: Article
• Language: English
• Subjects: acid hydrolysis; Adsorption; beta-glucuronidase; Brassica - chemistry; cabbage; cellulose; Chemical Precipitation; Enzyme Activation - drug effects; enzyme activity; Escherichia coli; Escherichia coli - enzymology; esterification; Glucuronidase - metabolism; hydrochloric acid; Hydrolysis; ion exchange chromatography; Pectins - chemistry; Pectins - metabolism; Pectins - pharmacology; Peptide Fragments - pharmacology; Plant Proteins - chemistry; Plant Proteins - metabolism; Plant Proteins - pharmacology; protein content; trichloroacetic acid
• ISSN: 0021-8561; 1520-5118
• DOI: 10.1021/jf305537x

We previously demonstrated that pectin–protein complex (PPC) isolated from white cabbage adsorbs the β-glucuronidase (βG) enzyme of E. coli. Concurrently, we discovered a significant increase in βG activity in the presence of PPC. The aim of this study is to identify the structural components of PPC that are responsible for βG adsorption and activation. PPC was isolated from white cabbage using a saline solution containing hydrochloric acid (pH 1.5) at 37 °C for 4 h. PPC proteins were precipitated by aqueous 10% (m/v) trichloroacetic acid to yield the pectin–protein fractions PPC1 and PPC2. PPC was digested using 1,4-α-d-galacturonase, yielding the PPC6 fraction. Partial acid hydrolysis of PPC revealed the galacturonan fraction, PPC3, to be the core of the macromolecule. The purified PPC4 and PPC5 fractions were isolated from PPC by ion-exchange chromatography on DEAE-cellulose. βG activity and its adsorption in the PPC fractions were studied in vitro. Crystalline cellulose was used as a control. This study found that the PPC3 fraction (the galacturonan core) does not adsorb βG and does not affect its activity. The adsorption of βG in the PPC samples is inversely proportional to the degree of methyl esterification of its carbohydrate component. The PPC4 and PPC5 fractions adsorb the highest proportion of βG (51.2% and 54%, respectively). The stimulation of βG enzyme activity is directly proportional to the protein content of the PPC sample. The PPC and PPC1 samples have the greatest ability to increase βG activity (57.6% and 52.1%, respectively).