Cross-linked Pectin Research Articles

Morphine Metabolism in the Opium Poppy and Its Possible Physiological Function: BIOCHEMICAL CHARACTERIZATION OF THE MORPHINE METABOLITE, BISMORPHINE

We identified a novel metabolic system of morphine in the opium poppy (Papaver somniferum L.). In response to stress, morphine is quickly metabolized to bismorphine consisting of two morphine units, followed by accumulation in the cell wall. This bismorphine binds predominantly to pectins, which possess high galacturonic acid residue contents, through ionical bonds. Our newly developed method using artificial polysaccharides demonstrated that bismorphine bridges are formed between the two amino groups of bismorphine and the carboxyl groups of galacturonic acid residues, resulting in cross-linking of galacturonic acid-containing polysaccharides to each other. The ability of bismorphine to cross-link pectins is much higher than that of Ca2+, which also acts as a cross-linker of these polysaccharides. Furthermore, we confirmed that cross-linking of pectins through bismorphine bridges leads to resistance against hydrolysis by pectinases. These results indicated that production of bismorphine is a defense response of the opium poppy. Bismorphine formation is catalyzed by anionic peroxidase that pre-exists in the capsules and leaves of opium poppies. The constitutive presence of morphine, together with bismorphine-forming peroxidase, enables the opium poppy to rapidly induce the defense system.

Purification of Muscle Enzymes by Pseudoaffinity Chromatography

ABSTRACT A method based on pseudoaffinity chromatography has been developed for the separation of lactate dehydrogenase (LDH), pyruvate kinase (PK) and aldolase from rabbit muscle extract using cross-linked guar (CLG) and cross-linked pectin (CLP) as the matrices, and dyes as the ligands. Screening of several dyes revealed that dyes No. 1014 and No. 1015, immobilized on CLG and CLP displayed a higher affinity for LDH and PK. Aldolase was not retained on any of the dye columns. It was observed that 1014-CLP and 1014-CLG columns retained 90% and 55% LDH activities, respectively, whereas 1015-CLP and 1015-CLG retained 83% LDH and 72% PK. A coupled-column system comprising 1014-CLP and 1015-CLP or 1014-CLG and 1015-CLG could separate LDH, PK, and aldolase from a mixture of these enzymes, as well as from rabbit muscle extract. Enzymes were found to be homogeneous on polyacrylamide gel electrophoresis. The method has been found to be simple and economical.

Developmental and ripening-related effects on the cell wall of apricot (Prunus armeniaca) fruit

Alcohol-insoluble residues (AIRs) were prepared from apricots at six stages during development/ripening on the tree. To investigate the changes in cell wall polymers, and in particular those affecting pectic polysaccharides, the AIR preparations were sequentially extracted with water, cyclohexane-trans-1,2-diamine-N,N,N′,N′-tetraacetate (CDTA) and Na2CO3. A significant proportion of initially Na2CO3-soluble pectic polysaccharides became water- and CDTA-soluble during the ripening process. In terms of composition, a significant decrease in galactose and uronic acid content was detected in all the extractions, whereas the percentage of arabinose increased in both water and CDTA-soluble polymers but decreased in the Na2CO3-extracted polysaccharides. The ability of pectic polysaccharides to cross-link was diminished during ripening due to an overall increase in the concentration of Na+ or K+ associated with the AIRs. This was accompanied by a decrease in the amounts of Ca2+ and Mg2+. The decrease in pectic galactans and the inhibition of pectin cross-linking detected within the pectic backbone are probably linked to the softening process observed during apricot ripening. © 1998 SCI.

Developmental and ripening‐related effects on the cell wall of apricot (Prunus armeniaca) fruit

Alcohol-insoluble residues (AIRs) were prepared from apricots at six stages during development/ripening on the tree. To investigate the changes in cell wall polymers, and in particular those affecting pectic polysaccharides, the AIR preparations were sequentially extracted with water, cyclohexane-trans-1,2-diamine-N,N,N′,N′-tetraacetate (CDTA) and Na2CO3. A significant proportion of initially Na2CO3-soluble pectic polysaccharides became water- and CDTA-soluble during the ripening process. In terms of composition, a significant decrease in galactose and uronic acid content was detected in all the extractions, whereas the percentage of arabinose increased in both water and CDTA-soluble polymers but decreased in the Na2CO3-extracted polysaccharides. The ability of pectic polysaccharides to cross-link was diminished during ripening due to an overall increase in the concentration of Na+ or K+ associated with the AIRs. This was accompanied by a decrease in the amounts of Ca2+ and Mg2+. The decrease in pectic galactans and the inhibition of pectin cross-linking detected within the pectic backbone are probably linked to the softening process observed during apricot ripening. © 1998 SCI.

Effect of preheating on potato texture.

Preheating potatoes at 50 to 80 degrees C has a firming effect on the cooked potato tissue. This effect is particularly pronounced at a preheating temperature of 60 to 70 degrees C followed by cooling. Several theories have been presented in the literature to explain this firming effect: retrogradation of starch, leaching of amylose, stabilization of the middle lamellae and cell walls by the activation of the pectin methylesterase (PME) enzyme, and by the release of calcium from gelatinized starch and the formation of calcium bridges between pectin molecules. Most probably, none of these theories alone can explain the phenomenon and more than one mechanism seems to be involved. Some of these mechanisms seem to be interdependent. As an example, calcium could be considered as a link all the way through release after starch gelatinization to cross-linking pectin substances in the cell wall and the middle lamellae, which has been demethylated by the PME enzyme. More research and "clear cut" experiments are needed in order to elucidate the role of each mechanism, especially which of them is the main contributor to the process of firming. Most probably, the calcium-pectin-PME mechanism plays a secondary role, that is, it only retards the collapse of the tissue structure that would otherwise occur during the final heating without preheating, and it is not the main factor of firmness.

Effect of Macerase, Oxalic Acid, and EGTA on Deep Supercooling and Pit Membrane Structure of Xylem Parenchyma of Peach

The object of this study was to determine if calcium cross-linking of pectin in the pit membrane of xylem parenchyma restricts water movement which results in deep supercooling. Current year shoots of ;Loring' peach (Prunus persica) were infiltrated with oxalic acid or EGTA solutions for 24 or 48 hours and then either prepared for ultrastructural analysis or subjected to differential thermal analysis. The effect of 0.25 to 1.0% pectinase (weight/volume) on deep supercooling was also investigated. The use of 5 to 50 millimolar oxalic acid and pectinase resulted in a significant reduction (flattening) of the low temperature exotherm and a distinct swelling and partial degradation of the pit membrane. EGTA (10 millimolar) for 24 or 48 hours shifted the low temperature exotherm to warmer temperatures and effected the outermost layer of the pit membrane. A hypothesis is presented on pectin-mediated regulation of deep supercooling of xylem parenchyma.

EFFECT OF CALCIUM CHELATORS ON WALL STRUCTURE AND DEEP SUPERCOOLING OF XYLEM PARENCHYMA OF PEACH

The pit membrane of xylem parenchyma of peach plays an important role in deep supercooling. Enzyme hydrolysis of xylem tissue indicated that the pit membrane is rich in pectin. The objective of the present study was to determine if removal of calcium from the cell wall would effect deep supercooling by loosening the cell wall. Current year shoots of `Loring' peach were infiltrated with oxalic acid, EGTA, or sodium phosphate buffer for 24-48 hours and then prepared for either ultrastructural analysis or differential thermal analysis. The use of 5-50 mM oxalic acid resulted in a distinct reduction in the size of the low-temperature exotherm (LTE) with increasing concentration. Oxalic acid also produced a loosening and swelling of the pit membrane. The use of EGTA (100 mM) or NaP04 (150 mM) produced only a slight shift in the LTE to warmer temperatures when compared to fresh tissues. Heat treatments (30-100°C) also resulted in a gradual shift of the LTE to warmer temperatures. The data indicate that cross-linking of pectins may play a role in defining the pore structure of the pit membrane and that this area of the cell wall plays an integral role in deep supercooling of peach wood.