A novel GH6 cellobiohydrolase from Paenibacillus curdlanolyticus B-6 and its synergistic action on cellulose degradation

We recently discovered a novel glycoside hydrolase family 6 (GH6) cellobiohydrolase from Paenibacillus curdlanolyticus B-6 (PcCel6A), which is rarely found in bacteria. This enzyme is a true exo-type cellobiohydrolase which exhibits high substrate specificity on amorphous cellulose and low substrate specificity on crystalline cellulose, while this showed no activity on substitution substrates, carboxymethyl cellulose and xylan, distinct from all other known GH6 cellobiohydrolases. Product profiles, HPLC analysis of the hydrolysis products and a schematic drawing of the substrate-binding subsites catalysing cellooligosaccharides can explain the new mode of action of this enzyme which prefers to hydrolyse cellopentaose. PcCel6A was not inhibited by glucose or cellobiose at concentrations up to 300 and 100 mM, respectively. A good synergistic effect for glucose production was found when PcCel6A acted together with processive endoglucanase Cel9R from Clostridium thermocellum and β-glucosidase CglT from Thermoanaerobacter brockii. These properties of PcCel6A make it a suitable candidate for industrial application in the cellulose degradation process.

     

Cytoprotective and anti-diabetic effects of Derris reticulata aqueous extract

The current study was aimed to investigate pancreatic protective and anti-diabetic activities of the aqueous extract of Derris reticulata stem. First, we evaluated a cytoprotective potential of D. reticulata extract on alloxan-induced damage in vitro. Treatment with D. reticulata extract at the doses of 250 and 500 μg/ml significantly increased cell viability of the pancreatic β-cell line RINm5F after exposure of alloxan. The anti-hyperglycemic activity of D. reticulata extract was further studied in alloxan-induced diabetic rats. A significant reduction in blood glucose level along with an increase in body weight was observed in diabetic rats treated with D. reticulata extract at 250 mg/kg body weight for 15 days. Serum aspartate transaminase and alanine transaminase levels were also significantly decreased compared to diabetic control rats. In accordance with in vitro cytoprotective effect, histopathological examination revealed that pancreatic islet cells of the extract-treated diabetic rat were less damage than those of the untreated diabetic group. In order to find another possible mechanism of action underling hypoglycemic activity, the effect on glucose absorption was examined using everted sac jejunum. The results showed that D. reticulata extract suppressed glucose absorption from small intestine. To corroborate safety use of D. reticulata extract, acute oral toxicity was also conducted in rats. Our results showed that none of the tested doses (250, 500, 1,000, and 2,000 mg/kg) induced signs of toxicity or mortality after administration of the extract. The results suggested that D. reticulata extract possess anti-diabetic activity, which resulting from its pancreatic cytoprotective effect and inhibition of intestinal glucose absorption.