Research On Production Of Gastrodin Through Biotransformation By Armillaria Luteo-virens Sacc

Gastrodin is the important ingredients of Gastrodia elata Blume owing to its important pharmacological properties. In this thesis, the major research is focused on the optimization of biotransformation conditions for gastrodin from HBA, separation and purification of the products, and structural identification, and then carrying out the animal experiments to study its anti-inflammatory effection and immune function using the synthetic gastrodin, and finally investigating the characterisitcs of glucosyltransferase from the biotransformation system. This study will provide the basis for understanding the biotransformation mechanism and regulating the gastrodin production.The HPLC system used throughout this study consisted of two Waters 510 pumps, a sample injector (Rheodyne, Cotati, CA, USA) with a 20μL loop, and a Waters 996 photodiode array detector. The column used was a reversed-phase Symmetry C18 column (150 mm×3.9 mm i.d., 5 urn; Waters). The mobile phase was acetonitrile (solvent A)-0.02% phosphoric acid solution (solvent B) in the aso-gradient mode as follows: 95% A and 5% B. The flow rate was 0.5 mL/min, and the effluent was monitored at 220 nm. Routine sample calculations were made by comparison of the peak area with that of the standard curve.Single factor experiments and response surface methodology were applied to optimize the bioconversion conditions for gastrodin production. The optimized results showed that the optimum transformation conditions composed of 3 mg/mL HBA, 6.5 g/30 mL inoculums level, 1.5% Tween 80, pH 4.5, and transformation temperature at 28℃. The bioconversion kinetics model was simulated and constructed in this study.The glucosyltransferase enzyme for biosynthesis of gastrodin from HBA was preliminary studied. The optimum temperature of the enzyme reaction was 50℃, and the pH was 6.0. The enzyme was stable at less than 50℃, and pH 6.0～7.0. The transformed products were obtained and identified by extraction, chromatography and its analysis from ¹H, ¹³C-NMR. The bioactive functions of the synthetic gastrodin were examined in vivo design, which include anti-inflammatory and immune function. It was confirmed that the synthetic gastrodin had anti-inflammatory effection and immune stimulatory function.