Preparation Of Enteric Nattokinase Capsule And Hypoglycemic Soyfunctional Food

1-Deoxnojirimycin(DNJ)and nattokinase were effective drug for cardiovascular diseases and diabetes,respectively.However,the poor stability in acid environment and quick inactivation after entered into gastric acid environment make nattokinase less effective as oral drug.Encapsulation of nattokinase could improve its stability in acid environment,thus would improve its curative effect as oral drug.On other hand DNJ,which with complicated extraction process,gained much less in liquid fermentation.Nevertheless,the food which contained DNJ and produced by solid fermentation could omitted the extraction produce,and the DNJ production could improve as well.In this study,the nattokinase capsule was made,and polyglutamic acid and algin was employed as capsule wall.The encapsulation rate,release rate as well as the stability in acid environment was determined by our research.The optimum reagent formula to form nattokinase cupsule were as follows:polyglutamic acid 4%,algin 3%,ethanol 80%,and CaCl2 4%.The encapsulation rate was 64.84%.By the encapsulation of nattokinase,40.00%of nattokinase could maintain the biological activity in acid environment.In the stimulated intestinal environment,all the nattokinase could be released within 80 min.In addition,this encapsulation method was suitable for various concentration(ranged from 4 to 40 FU/mL)of nattokinase solution.Secondly B.amyloliquefaciens HZ-12 was used as producing strains to produced food contained DNJ,and soybean was used as the substrate during the solid fermentation process.DNJ concentration,anticoagulant activity and antioxidative activities of the soybean food were evaluated.The cultivation conditions for B.amyloliquefaciens HZ-12 was optimized and the results were as follows:the optimal temperature was 37?,the inoculum age was 9 h,the inoculum amount was 9%(v/v),and the optimal soybean volume was 70 g/250mL.Under the optimized condition,the maximum concentration of DNJ was 870.22 mg/kgdry weight.The analysis of anticoagulant activity and antioxidative activity showed that when the concentration of the fermented extracts is 40 mg/mL the anticoagulant activity reached 69.30%and DPPH radical-scavenging reach 82.21%at 20 mg/mL fermented extracts which level obtained 54.74%DPPH radical-scavenging in the non-fermented soybean.Finally the research of the effect to soybean flavor by stains those lack of phosphotransbutyrylase(PTB)gene and leucine dehydrogenase(LeuDH)achieved under the condition of Bacillus amyloliquefaciens HZ-12,B.amyloliquefaciens HZ-12/APTB,B.amyloliquefaciens HZ-12/?PTB?LeuD as producing strains,soybean as substrate.The DNJ concentration and the total isobutyric acid/isovaleric acid concentration of B.amyloliquefaciens HZ-12,?PTB mutant and ?PTB?LeuDH mutant was 710.12 mg/kg dry weight,701.31 mg/kg dry weight,493.25 mg/kg dry weight and 0.58 mg/kg dry weight,0.48 mg/kg dry weight,0.48 mg/kg dry weight,respectively.The results showed that the flavor of the production will be improved without the gene of PTB and LeuDH.