The Study Of Gene Clone,Expression,Purification And Characteristics Of Lactobacillus Acidophilus Feruloyl Esterase

Ferulic acid 4-hydroxy 3-methoxy cinnamic acid,the name derived from Ferula feotida was first isolated in 1866.Numerous studies focusing on the ferulic acid,act as an antioxidant,able to neutralize free radicals and reactive on oxygen species.The antioxidant activity of phenolic compounds,FA has been studied and associated with several positive health effects such as mainly anti-carcinogenic,antiallergic,antiinflammatory,neuro-protective and cardiovascular effects as antioxidant activity by derivatives their possible contributions to health beneficial effects,and their bioavailability in food matrices.High levels of FA are present in agriculture residues like vegetables,fruits,cereals,and coffee,found both in free and bound forms.In the agriculture statistics estimated consumption of FA in food intake around of 150-250 mg/day.In yield the endoxylanase concentration in ferulic acid content is increased.The yield of ferulic acid depends on agriculture residue xylanase,ferulic acid in china about 800 million tons/year capita.The raw wheat bran to de-starch bran with different unit/ml of ?-amylase dosage,10,20 and 30 with effect of different times 60,90 and 120 minutes the best result were record on 20U at 90 minutes 0.5%/mg of starch content were determined.The qualitative analysis of ferulic acid was determined by agriculture residue wheat bran with the use of destarch wheat bran by TLC method.The de-starch wheat bran ferment with the Lactobacillus acidophilus at 37? for 12 hr incubation,the feruloyl esterase was separated on TLC plate,the clear band of FAE was recorded at 0.65mg/ml.The high quantity of ferulic acid was obtained from the fermentation of Lactobacillus acidophilus was genetically engineered by bacterial fermentation,the optimum ratio of ferulic acid on physiological and biochemical parameters through the vegetative growth of Lactobacillus acidophilus,by the screening and to extract DNA,the enzyme cloned with pET 28a by the using primers.The digested purified FAE744 clone use for protein expression with the using of competent cell BL21 and induced by IPTG.The protein concentration was recorded 28.6mg/ml after 12 hr incubation period.The general parameters revealed of ferulic acid basic characterization was measured by Michaelis Menton.All basic characteristics were measured by replication;the optimum temperature was recorded on 25? while the thermos-stability was measured on 55?.The optimum pH was determining 7.0pH whereas the pH stability was recorded on pH 6.5 to 7.0.The catalyzed reaction of FAEs by protein engineering to increase the potential of industrial application for the molecular dynamic mutagenesis technologies was applied.The application of these enzymes by molecular mutagenesis and protein engineering technologies have been extensively applied to improve their properties evolved the mutant variants of FAE744 to generate mutant library by PCR with the higher activity than the wild type enzyme and improve the temperature,pH and solvent tolerance.