Jinggangmycin Study Microbial Transformation False Amino Sugars α- Glucosidase Inhibitor Preparation

Inhibitors of alpha-glucosidase play an important role in the treatment ofdiabetes. Valienamine, a potent alpha-glucosidase inhibitor and intermediates ofhypoglycemic drug synthesis, has great medicinal value and broad market prospects.Using the method of biotransformation to produce valienamine has become thecurrent research focus due to the complex process and excessive steps in chemicalsynthesis of valienamine. In this study, we use jinggangmycin (validmycin),abio-pesticides as raw material, producing valienamine through microbial degradationof Jinggangmycin, both method and yield were described.A method of valienamine production by hydrolysis of Acarbose withalkaline(sodium hydroxide) was described,732and DOWEX1x2ion exchange resinwere used to purify the raw production after hydrolysis to obtain the standardproduction of valienamine. Method of TLC and HPLC were also optimized foraccurate analysis. The best Mobile phase for TLC is1-PrOH-AcOH-H2O,4:1:1(volumeratio)；spots colored by1%Ninhydrin solution. Pre-treatment of derivation was alsotested and the condition of derivation was confirmed. Derivation reaction was carriedout with the reaction system of dimethylformamide solution of1%p-nitrofluorobenzene and1.5%triethylamine, under conditions of100℃of waterbath for60min to make this reation fully complet. The product of derivation wasanalyzed by HPLC and the condition for HPLC are as follows: Phenomenex GeminiC18column; acetonitrile-water (15:85); column temperature:30°C; flow rate:1ml/min; detection wavelength:398nm.20soil samples were collected from different regions of the the soil to screen astrain which is able to transform validamycin to valienamine and validamycin wasused as the only substrate of carbon source in the medium. After Shaking flaskcultures,2positive single colony were picked out in Preliminary screening from82microorganisms and Strain Vali-2was finally choosen because of its Relatively higher transform rates.Based on the analysis of morphological, physiological and biochemicalcharacteristics, Vali-2belongs to genus of Bacillus, and it is a Gram-negative bacteria.This strain could decompose nitrate, gelatin and glucose, but no gas production;citrate can not be utilized. This strain also produce amylase, catalase; methyl redreaction shows negative outcome, VP reaction outcome is uncertain. The resultsfrom Blast analysis of16S rDNA shows the strain Vali-2was identified as Bacillusoleronius.Optimal design for Shaking flask fermentation was studied to increase the yield.Result showed that the optimal medium compositions are: peptone1%, yeast extract0.5%,0.5%NaCl,0.7%K2HPO4,0.3%KH2PO4,0.01%MgSO4. The optimal cultureconditions are as follows:1%Jinggangmycin, initial pH of medium is8.0, incubationtime is4d, the fluid volume is50/250ml, substrate Jinggangmycin concentration is1%,The amount of inoculation is15%.