Search PubMed⌕ Search

Biomedical subjects

Rudrapatnam N Tharanathan

Publications and source records attributed to Rudrapatnam N Tharanathan.

11 recordsLinked to original sources

Free radical-induced chitosan depolymerized products protect calf thymus DNA from oxidative damage.

Low molecular weight chitosan (LMWC) and chitooligosaccharides (COs), obtained by persulfate-induced depolymerization of chitosan showed scavenging of OH. and O2.- radicals and offered protection against calf thymus DNA damage. Over 85% inhibition of free radicals and DNA protection were observed. LMWC (0.05 micromol) showed a strong inhibitory activity compared to COs (3.6 micromol). Further, LMWC showed calf thymus DNA condensation reversibly giving stability, as evident from CD, TEM and melting curves (Tm). A fluorescence study suggests the binding of LMWC in the minor groove, forming H-bonds to the backbone phosphates without distorting the double helix structure.

Animals↗

Crosslinked chitosan--preparation and characterization.

Chitosan undergoes radical-induced depolymerization in the presence of potassium persulfate at 60 degrees C, leading to extensive crosslinking of the fragmented chains on subsequent cooling at 4 degrees C. As a result, a possible conformational change leading to higher crystallinity, as evidenced by IR, X-ray and 13C NMR was observed.

Carbohydrate Conformation↗

Characterization of chito-oligosaccharides prepared by chitosanolysis with the aid of papain and Pronase, and their bactericidal action against Bacillus cereus and Escherichia coli.

Papain (from papaya latex; EC 3.4.22.2) and Pronase (from Streptomyces griseus; EC 3.4.24.31) caused optimum depolymerization of chitosan at pH 3.5 and 37 degrees C, resulting in LMMC (low molecular mass chitosan) and chito-oligomeric-monomeric mixture. The yield of the latter was 14-16% and 14-19% respectively for papain- and Pronase-catalysed reactions, depending on the reaction time (1-5 h). HPLC revealed the presence of monomer(s) and oligomers of DP (degree of polymerization) 2-6, which was also confirmed by matrix-assisted laser-desorption ionization-time-of-flight MS. Along with the chito-oligomers, the appearance of only GlcNAc (N-acetylglucosamine) in Pronase-catalysed chitosanolysis was indicative of its different action pattern compared with papain. Fourier-transform infrared, liquid-state 13C-NMR spectra and CD analyses of chito-oligomeric-monomeric mixture indicated the release of GlcNAc/GlcNAc-rich oligomers. The monomeric sequence at the non-reducing ends of chito-oligomers was elucidated using N-acetylglucosaminidase. The chito-oligomeric-monomeric mixture showed better growth inhibitory activity towards Bacillus cereus and Escherichia coli compared with native chitosan. Optimum growth inhibition was observed with chito-oligomers of higher DP having low degree of acetylation. The latter caused pore formation and permeabilization of the cell wall of B. cereus, whereas blockage of nutrient flow due to the aggregation of chito-oligomers-monomers was responsible for the growth inhibition and lysis of E. coli, which were evidenced by scanning electron microscopy analysis. The spillage of cytoplasmic enzymes and native PAGE of the cell-free supernatant of B. cereus treated with chito-oligomeric-monomeric mixture further confirmed bactericidal activity of the latter. Use of papain and Pronase, which are inexpensive and easily available, for chitosanolysis, is of commercial importance, as the products released are of considerable biomedical value.

Anti-Bacterial Agents↗

Mango ripening--chemical and structural characterization of pectic and hemicellulosic polysaccharides.

Ripening of mango is characterized by a gradual, but natural softening of the fruit, which is due to progressive depolymerization of pectic and hemicellulosic polysaccharides with significant loss of galactose, arabinose and mannose residues at the ripe stage. Structural characterization employing permethylation followed by GC-MS analysis, IR and 13C NMR measurements revealed the major CWS fractions of both unripe and ripe mangoes to be of variable molecular weights and having a 1,4-linked galactan/galacturonan backbone, which is occasionally involved in side chain branches consisting of single residues of galactose and arabinose or oligomeric 1,5-linked arabinofuranose residues linked through 1,3-linkages; whereas the major hemicellulosic fractions of unripe mango to be of xyloglucan-type having 1,4-linked glucan backbone with branching by non-reducing terminal arabinose and xylose residues.

Fruit↗

Chitooligosaccharides--preparation with the aid of pectinase isozyme from Aspergillus niger and their antibacterial activity.

An isozyme of pectinase from Aspergillus niger with polygalacturonase activity caused chitosanolysis at pH 3.5, resulting in low-molecular weight chitosan (86%), chitooligosaccharides (COs, 4.8%) and monomers (2.2%). HPLC showed the presence of COs with DP ranging from 2 to 6. Charcoal-Celite chromatography and re-N-acetylation of the COs followed by CD, IR, MALDI-TOF-MS and FAB-MS analyses revealed an abundance of chitobiose, chitotriose and chitotetraose. The COs-monomeric mixture showed a bactericidal effect towards Bacillus cereus and Escherichia coli more efficiently than native chitosan. Among the chitooligomers, the hexamer showed maximum antibacterial effect followed by the penta-, tetra-, tri- and dimers. Of the two monomers, only GlcN showed slight bacterial growth inhibition. SEM revealed bactericidal action patterns of COs-monomeric mixture towards B. cereus and E. coli.

Anti-Bacterial Agents↗

Starch--value addition by modification.

Starch is one of the most important but flexible food ingredients possessing value added attributes for innumerable industrial applications. Its various chemically modified derivatives offer a great scope of high technological value in both food and non-food industries. Modified starches are designed to overcome one or more of the shortcomings, such as loss of viscosity and thickening power upon cooking and storage, particularly at low pH, retrogradation characteristics, syneresis, etc., of native starches. Oxidation, esterification, hydroxyalkylation, dextrinization, and cross-linking are some of the modifications commonly employed to prepare starch derivatives. In a way, starch modification provides desirable functional attributes as well as offering economic alternative to other hydrocolloid ingredients, such as gums and mucilages, which are unreliable in quality and availability. Resistant starch, a highly retrograded starch fractionformed upon food processing, is another useful starch derivative. It exhibits the beneficial physiological effects of therapeutic and nutritional values akin to dietary fiber. There awaits considerable opportunity for future developments, especially for tailor-made starch derivatives with multiple modifications and with the desired functional and nutritional properties, although the problem of obtaining legislative approval for the use of novel starch derivatives in processed food formulations is still under debate. Nevertheless, it can be predicted that new ventures in starch modifications and their diverse applications will continue to be of great interest in applied research.

Food Handling↗

Non-specific depolymerization of chitosan by pronase and characterization of the resultant products.

Pronase (type XXV serine protease from Streptomyces griseus) efficiently depolymerizes chitosan, a linear beta-->1,4-linked polysaccharide of 2-amino-deoxyglucose and 2-amino-2-N-acetylamino-D-glucose, to low-molecular weight chitosans (LMWC), chito-oligomers (degree of polymerization, 2-6) and monomer. The maximum depolymerization occurred at pH 3.5 and 37 degrees C, and the reaction obeyed Michaelis-Menten kinetics with a Km of 5.21 mg.mL(-1) and Vmax of 138.55 nmoles.min(-1).mg(-1). The molecular mass of the major product, LMWC, varied between 9.0 +/- 0.5 kDa depending on the reaction time. Scanning electron microscopy of LMWC showed an approximately eightfold decrease in particle size and characterization by infrared spectroscopy, circular dichroism, X-ray diffractometry and 13C-NMR revealed them to possess a lower degree of acetylation, hydration and crystallinity compared to chitosan. Chitosanolysis by pronase is an alternative and inexpensive method to produce a variety of chitosan degradation products that have wide and varied biofunctionalities.

Acetylation↗

Low molecular weight chitosans--preparation by depolymerization with Aspergillus niger pectinase, and characterization.

The viscosity of a chitosan solution was rapidly lowered in the presence of pectinase from Aspergillus niger at pH 3.0 and 37 degrees C. The low molecular weight chitosans (LMWC) had a molecular weight in the range 20,000-5000 Da. Circular dichroism spectra showed a decrease in the segment of acetylated glucosamine units, whereas X-ray diffraction and CP-MAS 13C NMR indicated higher crystallinity and polymorphism in LMWC. The latter on thermal drying resulted in structural alterations, and yielded an insoluble product. FT-IR and X-ray diffraction showed no evidence of either Schiff's base linkage or any annealed polymorph. CP-MAS 13C NMR showed marked changes in the chain conformations of LMWC, which are believed to be responsible for its loss of solubility and functionality.

Acetylation↗

Chitin--the undisputed biomolecule of great potential.

Of the truly abundant polysaccharides in Nature, only chitin has yet to find utilization in large quantity. Chitin is the second most abundant natural biopolymer derived from exoskeletons of crustaceans and also from cell walls of fungi and insects. Chitin is a linear beta 1,4-linked polymer of N-acetyl-D-glucosamine (GlcNAc), whereas chitosan, a copolymer of GlcNAc (approximately 20%) and glucosamine (GlcN, 80%) residues, is a product derived from de-N-acetylation of chitin in the presence of hot alkali. Chitosan is, in fact, a collective name representing a family of de-N-acetylated chitins deacetylated to different degrees. Both chitin/chitosan and their modified derivatives find extensive applications in medicine, agriculture, food, and non-food industries as well. They have emerged as a new class of physiological materials of highly sophisticated functions. Their application versatility is a great challenge to the scientific community and to industry. All these are the result of their versatile biological activity, excellent biocompatibility, and complete biodegradability in combination with low toxicity. Commercial availability of high-purity forms of chitin/chitosan and the continuous appearance of new types of chitin/chitosan derivatives with more and more useful and specific properties have led to an unlimited R&D efforts on this most versatile amino polysaccharide, chitin to find new applications, which are necessary to realize its full potential. Incidentally, this too has become an environmental priority. No doubt, chitin is surely an undisputed biomolecule of great potential.

Animals↗

Carbohydrates--the renewable raw materials of high biotechnological value.

Carbohydrates are the potential biomolecules derived from nature. Their molecular diversity has led to a bewildering variety of species, structures and characteristics all performing a large array of functions of great significance. Biologically they are vital as message (immunological) carriers, physiologically they are useful as energy (nutritional) reserves, and technologically they are needed for altering the texture and consistency (functional) of foods. Recent advances in glycobiology have opened up a new understanding of the role of sugars in biology and medicine. Noncellulosic beta-(1-3)-linked D-glucans, a group of polysaccharides found as constituents of fungi, algae, and higher plants, exhibit many interesting properties, depending on their molecular conformation. They are excellent 'biological response modifiers' and show significant immunomodulatory activities. They elicit a variety of host defense biological responses, for example, potent antitumor activity. On the other hand, the mixed-linkage ((1-3/1-4)-beta-linked) glucans are important constituents of cereal cell walls, where they perform properties of physiological importance, such as water holding capacity, porosity, and plasticity, which are useful at different stages of growth/development of plants. Of late, carbohydrate-based therapeutics are becoming the promise against many chronic diseases of today and tomorrow. Some of the characteristic features, structural attributes, functional significance, and applications of a selected few carbohydrate species are the subject matter of this review.

Biotechnology↗

Food-derived carbohydrates--structural complexity and functional diversity.

Carbohydrates are biomolecules abundantly available in nature. They are found in bewildering types ranging from simple sugars through oligo- and polysaccharides to glycoconjugates and saccharide complexes, each exhibiting characteristic bio-physiological and/or nutritional functions both in in vivo and in vitro systems. For example, their presence or inclusion in food dictates the texture (body) and gives desirable customer appeal (satisfaction), or their inclusion in the diet offers beneficial effects of great therapeutic value. Thus, carbohydrates are integrally involved in a multitude of biological functions such as regulation of the immune system, cellular signaling (communication), cell malignancy, antiinfection responses, host-pathogen interactions, etc. If starch is considered the major energy storage carbohydrate, the gums/mucilages and other non-starch carbohydrates are of structural significance. The most investigated properties of starch are its gelatinization and melting behavior, especially during food processing. This has led to the development of the food polymer science approach, which has enabled a new interpretive and experimental frame work for the study of the plasticizing influence of simple molecules such as water, sugars, etc. on food systems that are kinetically constrained. Starch, although considered fully digestible, has been challenged, and starch is found to be partly indigestible in the GI tract of humans. This fraction of starch-resisting digestion in vivo is known as resistant starch (RS). The latter, due to its excellent fermentative capacity in the gut, especially yielding butyric acid is considered a new tool for the creation of fiber-rich foods, which are of nutraceutical importance. By a careful control of the processing conditions the content of RS, a man-made fiber, can be increased to as high as 30%. Arabinoxylans are the major endospermic cell wall polysaccharides of cereals. In wheat they are found complexed with ferulic acid esters, which after oxidative coupling in vivo mediated by H2O2 and peroxidases or even by photochemical means give cross linked diferuloyl derivatives. The latter confer strength and extensibility to the cell wall and offer resistance for digestibility by ruminants. They also help blocking of the ingress of pathogens. The ester bound ferulic acid after oxidation in vivo generates reactive oxygen species that contribute to the fragmentation of non-starch polysaccharides (hemicelluloses), and thereby reduces the product viscosity, a property seen during long-term storage of rice. In plant tissues, the arabinogalactans are implicated in such diverse functions as cell-cell adhesion, nutrition of growing pollen tubes, response to microbial infections, and also as markers of identity expressed in the terminal sequences of saccharide chains.

Carbohydrates↗