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.     

Partial purification and characterization of protease of Bacillus proteolyticus CFR3001 isolated from fish processing waste and its antibacterial activities

Bacillus proteolyticus CFR3001 isolated from fish processing wastes (both fresh water and marine) produced an alkaline protease. The optimum conditions for cell growth and protease production were 37 degrees C, 96 h, agitation speed of 100 rpm and medium pH 9. The partially purified protease obtained from had specific activity of 22.05 at 37 degrees C was active between 40 degrees C and 50 degrees C and lost >20% of its activity around 60 degrees C. Its molecular weight was approximately 29 kDa and it inhibited the growth of several pathogenic organisms such as Escherichia coli, Listeria monocytogenes, Bacillus cereus and Yersinia enterocolytica. The scanning electron microscopy (SEM) studies revealed that the protease produced by B. proteolyticus CFR3001 lysed the cells of these pathogenic bacteria.     

Heterogeneity of ciguatoxins extracted from fish caught in the French Antilles

Ciguatoxin-like substances were extracted from the viscera or the flesh of eight Caribbean fish species, including small invertebrate feeders and large carnivores. The had similar properties, i.e. pharmacological action, solubility, chromatographic behaviour on silicic acid or Sephadex LH 20 column, stability in a weak acid solution and instability in alkaline medium. However, Florisil column and thin-layer chromatography showed different ciguatoxins whose number depended on tissue or species but not on fish trophic level. Less polar ciguatoxins appeared in salted and dried flesh. Thus, fish ciguatoxins are believed to be closely related substances, possibly changing in structure according to particular experimental conditions.     

Preparation and partial characterization of collagen sheet from fish (Lates calcarifer) scales

Fish scales, which are hitherto discarded as waste, were collected and cleaned thoroughly. The scales were hydrolyzed under controlled acidic conditions, neutralized and made in to a sheet, i.e., fish scale collagen sheet (FCS). The FCS was characterized for its infrared spectroscopy (IR), thermo-gravimetric analysis (TGA), scanning electron microscopy (SEM), and mechanical properties. The IR study has shown that the sheet contains both organic and inorganic phases revealing that the scales are partially deminaralized. The tensile strength of FCS is enough if it is used as a wound dressing material. The SEM studies have shown that FCS is porous and exhibited fibrous nature.     

Isolation and characterization of glycosaminoglycans from atria of the human heart

• 1.1. Five glycosaminoglycans were isolated from tryptic digest of atria of the human heart and were assayed by determining the carbohydrate content of materials.
• 2.2. Separation of these 5 polymers was achieved by Dowex 1 × 2 column chromatography.
• 3.3. They were identified as hyaluronic acid, heparan sulfate, chondroitin-4-sulfate, dermatan sulfate and chondroitin-6-sulfate, respectively.

     

Isolation and characterization of glycosaminoglycans from the Furth murine mastocytoma.

New methods for isolation and fractionation by partition are described and compared with existing techniques. Substantially purer products were isolated by partition as compared to precipitation with hexadecylpyridinium chloride. The glycosaminoglycans isolated fron Furth murine mastocytoma tumor were found to consist of 78-80% heparin, 12-13% chondroitin sulfate, and 8-9% hyaluronate. Dermatan sulfate was not detected. Two heparin-like glycosaminoglycans could be isolated by partition fractionation in the phase system 1-butanol/aqueous NaCl containing hexadecylpyridinium chloride. The composition of one was typical of heparins. However, the other glycosaminoglycan contained only 0.47 moles N-sulfate/mole uronate, but had electrophoretic and partition properties characteristic of heparin.     

Vegetalising factor extracted from the fish swimbladder and tested on presumptive ectoderm ofTriturus embryos

Summary Vegetalising factor was isolated from swimbladder of crusian carp (Carassius auratus) by solubilishing with 8 M urea the precipitate obtained after digesting the swimbladder with collagenase. The urea-soluble fraction vegetalised isolated presumptive ectoderm ofTriturus gastrula and produced both undifferentiated mesodermal and endodermal cells. Brief heating of the fraction changed its capacity to produce organised mesodermal tissues, such as notochord and somite, and the frequency of induction of undifferentiated cells was reduced. By inserting the urea-soluble fraction into the blastocoel of an early gastrula, embryos without epidermis were obtained. Some of the embryos consisted of undifferentiated mesodermal and endodermal cells, but in the remaining ones small fragments of notochord, small numbers of somites and pronephros developed, enclosed by endodermal cells.     

Further purification and partial characterization of the rat lung cytoplasmic factors modulating adenylate cyclase activity in plasma membrane

Summary The adult rat lung cytoplasm contains some factors which markedly stimulate adenylate cyclase activity in plasma membranes (Nijjar, M. S. Biochim. Biophys. Acta 584:43–50, 1979). Adenylate cyclase activator (ACA) was purified from rat lungs by DEAE-cellulose chromatography, preparative isoelectric focusing and by repeated high-performance liquid chromatography on a Sepharogel TSK 2000SW column. The final preparation showed about 200 fold purification in ACA activity over the original lung supernatant, and appeared to be homogeneous on the basis of its migration into a single band on SDS-polyacrylamide gel electrophoresis, and co-elution of ACA activity with protein from a gel exclusion column. ACA is an acidic (pl 4.8 ± 0.1), heat labile, monomeric protein of 40000 ± 2000 dalton molecular weight, and does not resemble calmodulin.