High Performance Preparation Of Gallic Acid From Galla Chinensis And Its Inhibition Function On α-Aamylase

Gallunt is the formation of cystic gall that pemphigidae aphids fimilines in thier host plants such as Sumac tree.Gallunt is rich in gallic acid,and it is one of the major resource production and preparation of gallic acid.Raw materials through the acid hydrolysis, alkaline hydrolysis and fermentation and then solven extraction or chromatography are the primary ways to prepare gallic acid in current industry.But the problems of low efficiency, low yield, environmental pollution, and the fact of prepared gallic acid is usually used as chemical raw materials, making the application potential not fully realized.This paper systematicly study the enzymatic degradation of gallict anninst hrough the system,high tannin enzyme microbial screening,macroporous resin column chromatography purification gallic acid and a-amylase in hibition effect.to establish an efficient method for the preparation of gallic acid.It proves mechanism and performance that gallic acid inhibition effect ofa-amylase initially,and it provides a scientific basis of the function of Gallunt gallic acid that efficient preparation and development to diabetes drug.The main findings are as follows:(1) Establishing the optimization technology of tannase enzymatic degradation enzyme gallunt. It is based on the study of enzymatic hydrolysis temperature, enzyme concentration,the enzymatic reaction time and pH enzymatic degradation gallunt preparation gallic acid by Tannin enzyme.and optimize the technology of enzymatic degradation gallunt preparation gallic acid by Tannin enzyme via response surface methodology.The results showed that the regression equation of the yield of gallic acid on the reaction temperature (A), enzyme concentration (B), reaction time (C), reaction pH (D) quadratic polynomial coding is:Y=-209.037+4.10378A+35.1799B+9.75150C+14.9282D-0.048273A2-2.33787B2-0.652825C2-1.05987D2-0.00275AB+0.02195AC-0.050025AD-0.575675BC-0.221238BD+0.137313DC.The optimal parameters is the enzyme concentration technology16.26U/g, the enzymatic reaction time is5.88h, the reaction system pH5.78,enzymatic hydrolysis temperature40.70℃. Under the conditions of this parameter, the yield of gallic acid is59.32%.(2)Screening a strains of microorganisms of a kind of high-yielding tannase.It adopts the method of combining flat transparent circles and solid fermentation.to isolated a strain that produce higher enzyme activity named the9th from soil.and it’s initial activity was 21.537U/mL.By colony morphology and ITS sequence analysis,the strain Harzianum is identified as the9th.The further study of impact of the fermentation temperature, time, and moisture content of the medium gallic on the production of enzyme activity, shows that on the conditions that fermentation time3d, fermentation temperature is33℃, gallic content is under20%and50%moisture content, the9th strains of producing the enzyme activity reaches25.96U/mL.(3)Studies on separating the Gallic acid from Gallunt by macroporous resins. The research compared eleven different kinds of macroporous resins through the absorption and desorbtion capacity of Gallic acid which were NKA-2、LS303、HZ816、HP-20、XAD-16、 HPD700、HPD500、S-8、AB-8、D101and D301,and the best processing parameters were optimized based on the adsorption and desorption results. The results indicated that NKA-2was the best resin. Its rate of the static absorbtion was94.856mg/g and the rate of desorption was77.26%. An best yield parameter with sample concentration as5mg/mL,sample volume as5BV,sample flow rate as3BV/h, elution ethanol concentration as70%, elution flow rate as2.0BV/h, elution volume as2.5BV was built and applied to separate gallic acid,in this conditiontion the result was obtained that the yield was85.04%and the purity was87.10%..An best purification parameter with sample concentration as7mg/mL, sample volume as7BV, sample flow rate as2BV/h, elution ethanol concentration as70%, elution flow rate as3.0BV/h, elution volume as2.5BV was built and applied to separate gallic acid,in this condition the result was obtained that yield was78.84%and the purity was90.97%.(4) Initially proving the mechanism and performance of the gallic acid inhibition of a-amylase via vitro methods. The results show that when gallic acid’s concentration reaches15mg/mL, and it’s inhibition rate of a-amylase activity reaches95.39%, the gallic acid inhibition of a-amylase is reversible inhibition.