Fermented The Residue From Wheat Alcohol Processing And Research On Antioxidant & Immune Activity Of Small Peptide

The residues from wheat alcohol processing have high content of moisture,bulky,bad smell and intractable.It has a high content of crude protein,Therefore,it is a waste of resources and environmental pollution if the residues were discarded.Solid-state fermentation not only make it harmless,but also realize recycling use.At the same time,bioactive peptides were increased in fermented products.In this study,the fermentation process was optimized,then separate and extract the small peptide mixtures,last its antioxidant and immune activity was tested.1 Fermented the residues from wheat alcohol processing for forage.Bacillus licheniformis D-1 and subtilis KG109 were used to ferment the residues.The fermented products were compared with the samples from Anhui Ruifuxiang food company koji fermenting.The amounts of inoculum in D-1 and KG109 set two gradients for 3% and 6%,and each gradient set three levels for 50%,60% and 70%.The control was inoculum in10% koji and 50% moisture was kept.High moisture and high content of acid soluble proteins were used as optimal combination.The results showed that use Bacillus licheniformis D-1 as bacteria,3% added amount of bacteria,70% moisture content,resulting in the best product,the acid-soluble protein and crude protein contents were14.63% and 31.75%,both very significantly higher?P<0.01?.2 Purification and separation of small peptides from fermented residues of wheat alcohol processing.Optimization of small peptide extraction conditions using a single factor test and measuring crude extract of the peptide content.Through the results,at30 ?,water ratio of 1:6,extract 15 min can get optimum results.Small peptide isolated and extracted using Sephadex G-15,optimized separation conditions,and ultimately five different molecular weight peptides,is determined by the standard curve,named octapeptide,heptapeptide,pentapeptide,tetrapeptide and tripeptide.3 Antioxidant Activity in Vitro and in Vivo & Cell Immune Activity Analysis of Small Peptides from Fermentd Residues of Wheat Alcohol Processing.The DPPH scavenging activity,hydroxyl radical scavenging activity and its total reducing capacity of five small peptides were measured in vitro.To determined antioxidant activity in vivo by mice gavaged tests,112 4-week-old healthy female Kunming mice were randomly divided into 14 groups.Mice of the control were fed with normal diet + saline and that of test 1-6were fed with normal diets + pentapeptide or heptapeptide at different doses?4,8,and 12mg?.Mice of the control were fed with high-fat diet + saline and that of test 7-12 were fed high-fat diets + pentapeptide or heptapeptide at different doses?2,4,and 8 mg?.After continuous intragastric 28 d,mice of normal control and test 1-6,the levels of malondialdehyde?MDA?in serum and liver,and the activity of the enzymes superoxide dismutase?SOD?,catalase?CAT?and glutathione peroxidase?GSH-Px?in liver were deternined.CD₃⁺?CD₄⁺and CD₈⁺in spleen were tested by flow cytometer.Mice of normal control,high-fat control and test 7-12 were made tissue homogenates from the duodenum,jejunum and ileum,then the levels of MDA,the activity of the enzymes SOD,CAT and GSH-Px were tested.The results showed that DPPH radical scavenging activity of 1mg/mL the peptide with 5 or 4 amino acid residues was about 80%,heptapeptide or tripeptide was 48% and 31%,but the peptide with 8 amino acid residues had no activity of scavenging the DPPH radical.In addition,the hydroxyl radical scavenging rate of 0.4mg/mL peptide with 4 amino acid residues and 0.8 mg/mL the crude extract was above95%,and the hydroxyl radical scavenging rate of 0.8mg/mL peptide with 5 or 7 amino acid residues was above 90% and 80%.While tripeptide was lower and octapeptide does not have the ·OH scavenging activity.Total reducing power of 5mg/mL crude extract and tetrapeptide was similar with 0.5mg/mL glutathione.And in mice fed small peptides,MDA content was reduced significantly?P < 0.05?.SOD,CAT and GSH-Px activity in liver,duodenum,jejunum and ileum were increased significantly.CD₄⁺/CD₈⁺ ratio was signifitcantly increased?P < 0.05?.Experimental results showed that tetrapeptide,pentapeptide and heptapeptide had strong antioxidant ability in vitro.pentapeptide and heptapeptidel have antioxidant capacity in vivo and can enhance the cell immune activity,also can inhibit the oxidative damage caused by high-fat diets in intestinal in the mice.