Van Bon Nguyen

Production and purification of a fungal chitosanase and chitooligomers from Penicillium janthinellum D4 and discovery of the enzyme activators

Chitosanases have received much attention because of their wide range of applications. Although most fungal chitosanases use sugar as their major carbon source, in the present work, a chitosanase was induced from a squid pen powder (SPP)-containing Penicillium janthinellum D4 medium and purified by ammonium sulphate precipitation and combined column chromatography. The purified D4 chitosanase exhibited optimum activity at pH 7-9, 60°C and was stable at pH 7-11, 25-50°C. The D4 chitosanase that was used for chitooligomers preparation was studied. The enzyme products revealed various chitooligomers with different degrees of polymerisation (DP) from 3 to 9, as determined by a MALDI-TOF mass spectrometer, confirming the endo-type nature of the D4 chitosanase. D4 chitosanase activity was significantly inhibited by Cu(2+), Mn(2+), and EDTA. However, Fe(2+) activated or inhibited D4 chitosanases at different concentrations. The D4 chitosanase was also activated by some small synthetic boron-containing molecules with boronate ester side chains.

Biosynthesis of α-Glucosidase Inhibitors by a Newly Isolated Bacterium, Paenibacillus sp. TKU042 and Its Effect on Reducing Plasma Glucose in a Mouse Model

Paenibacillus sp. TKU042, a bacterium isolated from Taiwanese soil, produced α-glucosidase inhibitors (aGIs) in the culture supernatant when commercial nutrient broth (NB) was used as the medium for fermentation. The supernatant of fermented NB (FNB) showed stronger inhibitory activities than acarbose, a commercial anti-diabetic drug. The IC50 and maximum α-glucosidase inhibitory activities (aGIA) of FNB and acarbose against α-glucosidase were 81 μg/mL, 92% and 1395 μg/mL, 63%, respectively. FNB was found to be strongly thermostable, retaining 95% of its relative activity, even after heating at 100 °C for 30 min. FNB was also stable at various pH values. Furthermore, FNB demonstrated antioxidant activity (IC50 = 2.23 mg/mL). In animal tests, FNB showed remarkable reductions in the plasma glucose of ICR (Institute of Cancer Research) mice at a concentration of 200 mg/kg. Combining FNB and acarbose enhanced the effect even more, with an added advantage of eliminating diarrhea. According to HPLC (High-performance liquid chromatography) fingerprinting, the Paenibacillus sp. TKU042 aGIs were not acarbose. All of the results suggest that Paenibacillus sp. TKU042 FNB could have potential use as a health food or to treat type 2 diabetes.