Delineating the inherent functional descriptors and biofunctionalities of pectic polysaccharides.

Pectin is a plant-based heteropolysaccharide macromolecule predominantly found in the cell wall of plants. Pectin is commercially extracted from apple pomace, citrus peels and sugar beet pulp and is widely used in the food industry as a stabilizer, emulsifier, encapsulant, and gelling agent. This review highlights various parameters considered important for describing the inherent properties and biofunctionalities of pectins in food systems. These inherent descriptors include monosaccharide composition, galacturonic acid content, degree of esterification, molecular weight, structural morphology, functional group analysis, and functional properties, such as water and oil holding capacity, emulsification, foaming capacity, foam stability, and viscosity. In this study, we also delineate their potential as a nutraceutical, prebiotic, and carrier for bioactive compounds. The biofunctionalities of pectin as an anticancer, antioxidant, lipid-lowering, and antidiabetic agent are also conceptually elaborated in the current review. The multidimensional characteristics of pectin make it a potential candidate for use in food and biomedical science.

Cr(VI) adsorption on the blends of Henna with chitosan microparticles: Experimental and statistical analysis.

In this work, the capability of blends of Henna with chitosan microparticles as novel adsorbent was studied in order to remove Cr(VI) ions. Response surface method (RSM) as a statistical technique was used to minimize number of experiments (26 runs), and optimize the operating conditions. In order to study Cr(VI) adsorption on the adsorbent, the effects of process parameters namely pH (2-9), initial Cr(VI) solution concentration (10-100 mg/L), Henna dose (0.1-1 g/L), chitosan microparticles dose (0.1-1 g/L) and contact time (20-150 min) were carefully considered. The adsorption did not change from pH 2 to 9 while it was reduced from 100% to 98.67% by growing Cr(VI) initial concentration. In addition, the adsorption was increased from 98.04% to 100% by increasing the adsorbent amount. The result of the isotherm study fitted well to Langmuir model (with R2 = 0.9826).

Immobilization of yeast inulinase on chitosan beads for the hydrolysis of inulin in a batch system.

An extracellular inulinase was partially purified by ethanol precipitation and gel exclusion chromatography from a cell free extract of Kluyveromyces marxianus. Partially purified inulinase exhibited 420 IU/mg specific activity and it was immobilized on chitosan beads. Activity yield of immobilized inulinase was optimized with glutaraldehyde concentration (1-5%), glutaraldehyde treatment time (30-240min), enzyme coupling-time (2-16h) and enzyme loading (5-30 IU) as functions. Under the optimized conditions maximum yield 65.5% of immobilized inulinase was obtained. Maximum hydrolysis of inulin 84.5% and 78.2% was observed at 125rpm after 4h by immobilized and free enzyme, respectively. A retention-time of 4h and 5h was found optimal for the hydrolysis of inulin under agitation (125rpm) by free and immobilized enzyme, respectively. The recycling of the developed immobilized biocatalyst was carried out after 5h of inulin hydrolysis in a batch system. The developed immobilized biocatalyst was successfully used for the hydrolysis of inulin for 14 batches. This is the first report on the immobilization of yeast inulinase on chitosan beads for the hydrolysis of inulin in a batch system.

Welan gum: microbial production, characterization, and applications.

Microbial exopolysaccharides are of high molecular weight, environment friendly valuable natural polymers, having applications in diverse areas such as food industry, pharmaceutical industry, cement systems and cosmetics industries. Welan gum, a microbial product holds an important place among the exopolysaccharides due to its novel properties and potential applications. The biopolymer welan gum is synthesized by the fermentation process mainly by the Alcaligenes sp., and is composed of polymer of tetrasaccharide backbone chain containing l-mannose, l-rhamnose, d-glucose, and d-glucuronic acid. It acts as a thickening, suspending, binding, emulsifying, stabilizing and viscosifying agent. It has important commercial applications in the cement systems. This review is primarily focused on the microbial production, purification, recovery, and the characterization based on the available published literature on the welan gum. Besides this, the biosynthesis and the various process factors affecting the production as well as properties of welan gum and its various applications have also been addressed.

Immobilization of ß-d-galactosidase from Kluyveromyces lactis on functionalized silicon dioxide nanoparticles: characterization and lactose hydrolysis.

ß-D-Galactosidase (BGAL) from Kluyveromyces lactis was covalently immobilized to functionalized silicon dioxide nanoparticles (10-20 nm). The binding of the enzyme to the nanoparticles was confirmed by Fourier transform-infrared spectroscopy (FTIR) and scanning electron microscopy (SEM). Functionalized nanoparticles showed 87% immobilization yield. Soluble and immobilized enzyme preparation exhibited pH-optima at pH 6.5 and 7.0, respectively, with temperature optima at 35 and 40°C, respectively. Michaelis constant (K(m)) was 4.77 and 8.4mM for free and immobilized BGAL, respectively. V(max) for the soluble and immobilized enzyme was 12.25 and 13.51 U/ml, respectively. Nanoparticle immobilized BGAL demonstrated improved stability after favoring multipoint covalent attachment. Thermal stability of the immobilized enzyme was enhanced at 40, 50 and 65°C. Immobilized nanoparticle-enzyme conjugate retained more than 50% enzyme activity up to the eleventh cycle. Maximum lactose hydrolysis by immobilized BGAL was achieved at 8h.

Molecular characterization and enzymatic hydrolysis of naringin extracted from kinnow peel waste.

Kinnow peel, a waste rich in glycosylated phenolic substances, is the principal by-product of the citrus fruit processing industry and its disposal is becoming a major problem. This peel is rich in naringin and may be used for rhamnose production by utilizing α-L-rhamnosidase (EC 3.2.1.40), an enzyme that catalyzes the cleavage of terminal rhamnosyl groups from naringin to yield prunin and rhamnose. In this work, infrared (IR) spectroscopy confirmed molecular characteristics of naringin extracted from kinnow peel waste. Further, recombinant α-L-rhamnosidase purified from Escherichia coli cells using immobilized metal-chelate affinity chromatography (IMAC) was used for naringin hydrolysis. The purified enzyme was inhibited by Hg2+ (1 mM), 4-hydroxymercuribenzoate (0.1 mM) and cyanamide (0.1 mM). The purified enzyme established hydrolysis of naringin extracted from kinnow peel and thus endorses its industrial applicability for producing rhamnose.

Cell disruption optimization and covalent immobilization of beta-D-galactosidase from Kluyveromyces marxianus YW-1 for lactose hydrolysis in milk.

beta-D-galactosidase (EC 3.2.1.23) from Kluyveromyces marxianus YW-1, an isolate from whey, has been studied in terms of cell disruption to liberate the useful enzyme. The enzyme produced in a bioreactor on a wheat bran medium has been successfully immobilized with a view to developing a commercially usable technology for lactose hydrolysis in the food industry. Three chemical and three physical methods of cell disruption were tested and a method of grinding with river sand was found to give highest enzyme activity (720 U). The enzyme was covalently immobilized on gelatin. Immobilized enzyme had optimum pH and temperature of 7.0 and 40 degrees C, respectively and was found to give 49% hydrolysis of lactose in milk after 4 h of incubation. The immobilized enzyme was used for eight hydrolysis batches without appreciable loss in activity. The retention of high catalytic activity compared with the losses experienced with several previously reported immobilized versions of the enzyme is significant. The method of immobilization is simple, effective, and can be used for the immobilization of other enzymes.

WITHDRAWN: Nanomaterials Chemistry: Recent Developments and New Directions.

The Publisher regrets that this article is an accidental duplication of an article that has already been published in Int. J. Biol. Macromol, doi:10.1016/j.ijbiomac.2008.07.012. The duplicate article has therefore been withdrawn.

Immobilization of Candida rugosa lipase on magnetized Dacron: kinetic study.

Candida rugosa lipase has been covalently immobilized on ferromagnetic azide polyethyleneterepthalate (Dacron) with specific activity retention of 16% for 4-nitrophenyl palmitate and 24% for hydrolysis of triolein in hexane. The immobilized enzyme was more thermal stable than the soluble one, retaining 78.8% of the activity after 1 h at 60 degrees C. Also, this immobilized derivative was stable at the storage at 4 degrees C. It has been used 5 cycles for pNPP hydrolysis without loss of activity. Soluble and immobilized Candida rugosa lipase showed a Michaelian behavior for fatty acid 4-nitrophenyl esters and different apparent K(M) values: 0.110 mM and 0.124 mM (4-nitrophenyl palmitate - C16); 0.193 mM and 0.235 mM (4-nitrophenyl laurate - C12) and 0.206 mM and 0.119 mM (4-nitrophenyl butyrate - C4), respectively. The immobilized lipase was more efficient for catalyzing the hydrolysis of 4-nitrophenyl esters with short chain length fatty acid (4-NPB - C4) than soluble enzyme. The ferromagnetic Dacron-lipase derivative was able to catalyze the synthesis of triolein from glycerol and oleic acid with 50% of conversion after 72 h at 40 degrees C.

Applicability of pectate-entrapped Lactobacillus casei cells for L(+) lactic acid production from whey.

Lactic acid is a versatile organic acid, which finds major application in the food, pharmaceuticals, and chemical industries. Microbial fermentation has the advantage that by choosing a strain of lactic acid bacteria producing only one of the isomers, an optically pure product can be obtained. The production of L: (+) lactic acid is of significant importance from nutritional viewpoint and finds greater use in food industry. In view of economic significance of immobilization technology over the free-cell system, immobilized preparation of Lactobacillus casei was employed in the present investigation to produce L: (+) lactic acid from whey medium. The process conditions for the immobilization of this bacterium using calcium pectate gel were optimized, and the developed cell system was found stable during whey fermentation to lactic acid. A high lactose conversion (94.37%) to lactic acid (32.95 g/l) was achieved with the developed immobilized system. The long-term viability of the pectate-entrapped bacterial cells was tested by reusing the immobilized bacterial biomass, and the entrapped bacterial cells showed no decrease in lactose conversion to lactic acid up to 16 batches, which proved its high stability and potential for commercial application.

Encapsulated lactic acid bacteria for control of malolactic fermentation in wine.

The kinetics of both malolactic fermentation in Chardonnay wine by encapsulating Lactobacillus casei cells in pectate gel and lyophilized Oenococcus oeni culture has been carried out. The influence of acidity, sulfur dioxide content, and organic acid content on the malolactic activity of the bacteria has been controlled. Encapsulated bacteria degraded 30%, of malic acid in white wine, deacidifying it from pH 3.15 to 3.40, whereas the lyophilized culture degraded 48% of malic acid, deacidifying from pH 3.15 to 3.60. The degree of conversion of malic acid in wine by the encapsulated cells was twice as high as that obtained by the free Lactobacillus casei cells. The operational stability of calcium pectate gel capsules was 6 months. It has been proved that the encapsulated biocatalyst increases the rate of fermentation, and induces the fermentation to take place at high ethanol concentrations. The proposed encapsulated biocatalyst is an attractive material for industrial applications in continuous winemaking processes.

Isolation of a trypsin inhibitor from Echinodorus paniculatus seeds by affinity chromatography on immobilized Cratylia mollis isolectins.

A highly purified trypsin inhibitor was obtained from Echinodorus paniculatus when an extract prepared from E. paniculatus seed flour (25 gl(-1), with 0.1 M ammonium acetate buffer, pH 8.3, under agitation for 6 min at 28 degrees C) was chromatographed on Sephadex G-25 (12 mlh(-1)), followed by affinity chromatography on immobilized Cratylia mollis isolectins (Cra Iso 1,2,3-Sepharose). The column chromatography was performed at 24 degrees C; the matrix was washed (30 mlh(-1)) with 0.1 M sodium phosphate buffer, pH 7.4 or with the same buffer containing 0.2 M glucose, followed by application of inhibitor sample and elution with 0.015 M sodium borate buffer, pH 7.4, or 1.0 M NaCl. A purified fraction of inhibitor was obtained by gel filtration chromatography (GF-450/HPLC column). Trypsin inhibitory activity was eliminated when the inhibitor was treated with metaperiodate showing that the carbohydrate moiety was important for trypsin inhibition. Binding of inhibitor was also evaluated on immobilized concanavalin A (Con A-Sepharose) using previously described chromatographic conditions with results similar to Cra Iso 1,2,3-Sepharose chromatography.

Advanced wound care materials: developing an alginate fibre containing branan ferulate.

OBJECTIVE: This laboratory-based study set out to establish whether branan ferulate--a polysaccharide compound available in gels such as as Sterigel (SSL International, UK)--could be successfully added to the fibre of an alginate dressing to provide a superior wound-care dressing. METHOD: A wet extruder (Howden Engineering Services, UK) was used to produce the fibres. Researchers examined the effect of spinning-bath calcium chloride concentration (used as the coagulant in the fibre-spinning process) on the tensile and absorption properties of alginate fibres containing 25% w/w branan ferulate. Sodium alginate and branan ferulate were separately dissolved in de-ionised water and then stirred together to make up a 5% dope solution. The dope was then forced through a spinneret and into the calcium chloride bath, where fibres were precipitated and then carried over the first set of rollers. The fibres were washed in the next bath and drawn at different rates to enhance their tensile properties. At the 'winding-up' stage the filaments were either passed over a contact heater and into a furnace (dry pick-up) or transferred into acetone baths of increasing concentrations of 50, 80 and 100% aqueous acetate to remove the water (wet pick-up). The moisture content of standard alginate fibres and those with branan ferulate were measured and compared. Breaking load, tenacity and breaking extension, and the quantity of branan ferulate in the fibres were also measured. The process was repeated four times, using different strengths of calcium chloride. Using various draw ratios, the effects of liquid uptake were measured using water, saline and a solution called A (to mimic human blood and exudate-type fluids). RESULTS: Out of the four calcium chloride concentrations used, 1% concentration appeared to give the highest and most conclusive results for fibre-breaking load, tenacity and breaking extension. CONCLUSION: A 1% w/v concentration of calcium chloride provides the optimum conditions for achieving suitably strong fibres with adequate absorption capacity, while allowing the least branan ferulate loss during wash and post-production treatments. In a laboratory setting the product showed promise. Research is now needed in a clinical setting and for comparison with existing wound-care products.

Advances in enzymatic transformation of penicillins to 6-aminopenicillanic acid (6-APA).

This article elaborates on the important recent developments in the enzymatic transformation of penicillins to 6-aminopenicillanic acid (6-APA), which is the basic raw material for the industrial production of semisynthetic penicillins such as amoxycillin and ampicillin. Particular emphasis is placed on the improvements in purification, stability, and immobilization of the enzymes, (i.e. penicillin acylases) used for these transformations.

Immobilization of pneumococcal polysaccharide vaccine on silicon oxide wafer for an acoustical biosensor.

One the most important aspects of a biosensor is related to immobilization and maintenance of specific reference compounds on sensing surfaces. A method for the immobilization of polysaccharides to a silicon oxide surface intended for Surface Acoustical Waves (SAW) sensors is described. Silicon oxide is a hydrophobic inorganic support used for the fabrication of many electronic devices. The pneumococcal polysaccharide (PPS) vaccine is immobilized via Protein A after pre-treatment of the surface with hydrochloric acid. The effects of non-specific binding are discussed. The results indicate that the immobilization of PPS via Protein A increases the sensitivity of detecting Streptococcus pneumoniae antibodies in human sera and offers greater reproducibility of response compared with ELISA methods. The principles of this technique are simple and are applicable to the immobilization of many capsular polysaccharides.

Carbon dioxide transport by proteic and facilitated transport membranes.

Membrane separation of gases is governed by the permeability of each species across the membrane. The ratio of permeabilities yields the selectivity. Use of certain organic carriers in facilitated transport membranes and the CO2 converting enzyme carbonic anhydrase (CA) in proteic and facilitated transport membranes allows a dramatic increase in CO2 selectivity over other gases. CA has a low Km (9 mM), which we predicted would allow it to scavenge CO2 to very low partial pressures. Our goal was to determine if CA could remove CO2 from an environment at levels of 0.1% or less. Prior measurements of CO2 transport across thin supported liquid membranes showed that addition of CA enhanced CO2 flux by 3- to 100-fold. Proteic films use bifunctional reagents (e.g., glutaraldehyde) to cross-link the enzyme forming a gel. Bovine serum albumin (BSA) is often added for structural stability. Using such a preparation we examined the ability of proteic films to improve CO2 selectivity and to scavenge CO2 from a mixed gas stream. Proof-of-concept results, measured by mass spectrometry, showed a fivefold improvement in CO2 capture rate with maximal improvement at CO2 values of 1% partial pressure difference in the presence of 0 atm absolute difference. At 0.1% CO2 the membrane exhibited a 76% improvement over controls. At 0.3% CO2 the improvement is about threefold. CA proteic membranes exhibit selectivity for CO2 over oxygen and nitrogen in excess of three orders of magnitude. A CA-based proteic or facilitated transport membrane should readily achieve CO2 partial pressures of 0.05% under CELSS conditions. In addition to proteic membranes we are exploring direct immobilization of engineered CA to ultra-high-permeability teflon membranes. Site-directed mutagenesis was used to add functional groups while retaining full enzymatic activity. These results provide a basis for development of far more efficient CO2 capture proteic and facilitated transport membranes with increased selectivity to values closer to 100-fold at 1% CO2. The result will be CO2 selectivity at 0.1% on the order of 400-fold. These results exceed those obtained with other technologies.

Enteral feeding for the critically ill patient.

This article challenges nurses to examine their practice and procedure when establishing enteral feeding. The author suggests that discontinuation for 'complications' such as diarrhoea may not be appropriate, and recommends that nurses take more responsibility for patient nutrition.

Rheological studies on the solubilization by the surfactant SDS of complexes between three acidic polysaccharides and an organic base chloride.

Developments in the solubilization of complexes between the cationic polymer, poly(hexamethylenebiguanidinium chloride) (PHMBH+Cl-) and acidic polysaccharides are reported. It was discovered that the anionic detergent sodium dodecyl sulphate (SDS) is an excellent solubilizer for these complexes, enabling several multi-component systems to be studied. SDS itself was shown to interact with PHMBH+Cl- to form a highly viscous solution. Maximum viscosity was obtained with a SDS:PHMBH+Cl- molar ratio of 15.6. SDS:PHMBH+Cl- at this ratio served as a good solubilizer for the acidic polysaccharides (sodium alginate, sodium carboxymethyl cellulose, and xanthan), forming highly viscous fluids. The effect of temperature on the viscosity of these solutions was also examined.

Glycoproteins: a consideration of the potential problems and their solutions with respect to purification and characterisation.

Glycoproteins, as a class of biomolecules, present a number of differences in structure and function from non-glycosylated proteins. This makes their purification and characterisation a potentially daunting prospect. This article provides a brief summary of the occurrence, structure and composition of glycoproteins and the main areas where these factors may complicate the purification and characterisation process. There follows a review of the range of techniques available, an insight into the choices of ways forward in considering glycoprotein purification and characterisation. Appropriate prominence is given to lectins; their application to the purification, identification and characterisation of glycoproteins, and the major contribution they have made to the study of this class of biomolecules.