Characterization and enhanced production of prodigiosin from the spoiled coconut.

Many bacterial secondary products are bioactive substances that play an important role in biotechnology and pharmacology (e.g., as antibiotics or antitumor agents). Over the past few years interest in prodigiosin has been increased due to its promising anti-cancer activity. Prodigiosin is also of potential clinical interest because it is reported to have anti-fungal, anti-bacterial, anti-protozoal/anti-malarial, and immunosuppressive activity. Thus there is a need to develop a high-throughput and cost-effective bioprocess for the production of prodigiosin. In the present study, Serratia rubidaea was isolated from colored portion of a spoiled coconut and further it was authenticated by MTCC, India. The various parameters like temperature, pH, salt concentration, and precursors were optimized for the production of prodigiosin. We now report that the pigment production was higher in our isolated strain than S. marcescens. It was observed that prodigiosin binds with plastic, paper, and fibers and thus in near future, it can also be used as a natural dye.

Purification and properties of alpha-galactosidase from white-rot fungus Pleurotus florida.

alpha-Galactosidase was strongly induced in the white-rot fungus Pleurotus florida by arabinose than its natural substrates and was purified to homogeneity by acetone precipitation, ultrafiltration and DEAE-Sepharose chromatography. The enzyme was a monomeric protein with a molecular mass of approximately equal to 99 kDa, as revealed by native-PAGE and SDS-PAGE. alpha-Galactosidase was optimally active at 55 degrees C for the hydrolysis of p-nitrophenyl-alpha-galactopyranoside (PNPalphaG) and lost its 20% and 50% of original activity in 30 min at 60 degres C and 70 degrees C, respectively. The pH optimum of the enzyme was between 4.6 and 5.0. It was stable in a wide pH range (pH 4.0 to 9.0) at 55 degrees C for 2 h. The Ag+ and Hg2+ strongly inhibited the enzyme activity. Galactose, glucose, maltose and lactose also inhibited the enzyme activity, whereas N-bromosuccinimide treatment resulted in near total loss of acitivity. The Km and Vmax values of the enzyme for PNPalphaG were found to be 1.1 mM, and 77 micromol min(-1) mg(-1), respectively. alpha-Galactosidase immobilized in agar was more effective for the degradation of raffinose than in the sodium alginate. TLC results indicated its potential for the removal of raffinose and stachyose in soymilk.

Influence of sodium pentosan polysulphate and certain inhibitors on calcium oxalate crystal growth.

Calcium oxalate crystal growth and aggregation leads to the formation of renal calculi. It is known to be inhibited by several compounds both in vitro and in vivo conditions. The present study highlights the inhibitory potential of sodium pentosan polysulphate (SPP), a semi-synthetic glycosaminoglycan (GAG) on calcium oxalate crystal growth in vitro. Its efficacy was compared with those of known inhibitors like pyrophosphate, heparin and chondroitin-4-sulphate. Of the above compounds pyrophosphate was found to be the most potent inhibitor. Among the GAGs, SPP exhibited 80% inhibitory activity as compared to heparin. A lesser degree of inhibition was observed with chondroitin-4-sulphate.

Effect of sodium pentosan polysulphate on tissue lipids in control and glycollate treated rats.

Sodium pentosan polysulphate (SPP) has been reported to have a lipid lowering action. In addition it has been shown to prevent calcium oxalate crystallization in vitro and in vivo. Hence it was felt worthwhile to study the effect of SPP on altered tissue lipid levels, manifested in hyperoxaluric condition during glycollate feeding in rats. Elevated cholesterol with reduced phospholipid levels in both liver and kidney tissues, were the significant observations in the experimental animals. In addition total lipids were increased in the kidney. Administration of SPP to hyperoxaluric rats reduced tissue cholesterol and triglyceride levels significantly and raised the phospholipid levels in the tissues. Cholesterol ester synthetase (CES) and cholesterol ester hydrolase (CEH) activities were also assayed in the liver. Glycollate fed rats exhibited increased CES activity. SPP treatment had a lowering effect on CES but enhanced the CEH activity in both control and glycollate administered rats. The alterations may have a bearing in relation to calcium oxalate stone formation.

Alterations in some risk factors and urinary enzymes in urolithiatic rats treated with sodium pentosan polysulphate.

The effect of sodium pentosan polysulphate (SPP) was investigated in calcium oxalate stone forming rats with respect to the urinary excretion of certain risk factors and enzymes. Calcium oxalate stones were induced by feeding 3% w/w sodium glycollate to the rats. Urinary calcium, oxalate, phosphorus and uric acid levels were increased in stone formers. In contrast magnesium excretion was low in this group. SPP treatment lowered oxalate and calcium levels in both controls and experimental animals. Magnesium levels were increased moderately. Increased excretion of urinary enzymes--LDH, alkaline phosphatase, gamma-GT and beta glucuronidase--in calculogenic rats indicates membranuria and damage to proximal tubules during stone formation. Decreased pyrophosphatase activity was observed in glycollate fed rats. SPP treatment decreased the excretion of the above enzymes in the treated groups. Stone formers exhibited decreased LAP and fibrinolytic (urokinase) activities. SPP being associated with fibrinolytic properties, increased the activities of the above two enzymes to that of control levels in calculogenic rats.

Effect of L(+)-tartrate on some biochemical and enzymatic parameters in normal and glycollate treated rats.

Some biochemical and enzymatic constituents were determined in the small intestinal tract tissues of normal and sodium glycollate treated adult male rats. Alterations were observed with respect to certain lipids and carbohydrate fractions in the glycollate fed rats. DNA content was also elevated in this group. The functions of the cell membrane is likely to be affected as reflected in the levels of transport ATPases and orthophospho-hydrolases. The activities of the two marker enzymes in the intestinal brush border, namely alkaline phosphatase and leucylnaphthylamidase were reduced in the glycollate administered group. Administration of L(+)-tartrate, which is a mild laxative and has a regulatory influence on oxalate metabolism, lowered the activities of Na+, K(+)- and Ca(2+)-ATPases. There was a distinct lowering in the level of acid phosphatase in the tartrate treated rats.

Changes in serum lipids and lipoproteins in calcium oxalate stone forming rats treated with sodium pentosan polysulphate.

Sodium pentosan polysulphate (SPP) is reported to influence lipid metabolism, and since a relationship between lipids and stone disorder has been recognised, it was thought worthwhile to study the role of SPP in relation to serum lipids and lipoproteins in experimental calcium oxalate urolithiasis. Serum cholesterol, triglycerides and phospholipids were significantly increased in the glycollate treated group while free fatty acids showed only a mild increase. SPP treatment decreased the cholesterol and triglyceride levels in both controls and stone formers. In contrast, phospholipid and free fatty acid levels were increased in the two groups. In the calculogenic rats, the LDL/HDL cholesterol ratio showed no change, whereas with SPP administration there was a decrease in the treated groups. The observations are suggestive, that SPP treatment may prove beneficial in decreasing the blood lipid levels.

Biochemical changes in kidneys of normal and stone forming rats with sodium pentosan polysulphate.

The influence of sodium pentosan polysulphate was studied on the deposition of stone forming constituents along with certain enzymes in the renal tissue of experimentally induced urolithiatic rats. Calcium, oxalate and phosphorus levels were elevated in kidneys of lithogenic rats, while SPP administration reduced these levels to near control values. The elevation in kidney LDH was significant in the stone forming groups and SPP had minimal effect. Increases in the activities of Na+, K(+)-and Ca(2+)-ATPases in the calculogenic groups was lowered considerably with SPP treatment. Inorganic pyrophosphatase activity was reduced significantly in the calculogenic as well as in the drug treated groups. Leucine aminopeptidase was decreased in the calculogenic group. SPP treatment elevated the enzyme activity in the treated groups. Reduction in kidney oxalate with SPP may prove useful in the medical management of urolithiasis.