The Study Of Immune Activity And Development Functional Products Of Oratosquilla Oratoria Peptide

Marine bioactive peptide become a worldwide research hotpot in recent years,because of the features such as small molecular weight,high biological activity and low side effects.This paper mainly studies peptides of the Oratosquilla oratoria from enzymic extraction,then optimize the process of enzymic extraction,and after the separation and purification preliminary study on the immune activity of peptides.Basing on which,making peptides into capsule and establish a quality standard.All these will be the theoretical foundation in further study of the peptide from Oratosquilla oratoria.The main research results are as follows:1 The optimize of the enzymic extraction processIn order to determine the optimum enzymic extraction conditions of the immune active peptide from Oratosquilla oratoria,the preliminary exploration of enzymolysis conditions experiment has carried on with four single factors,which are the enzymic extraction's temperature,time,pH value and the concentration of trypsin,then using enzymatic hydrolysate influences the mice macrophage's cell activity to determine the appropriate level of single factor by MTT method.Basing on that,using the orthogonal experiment table design experiment to get the best conditions of enzymatic hydrolysis,and the result is:enzyme solution pH value of 9.5,enzymolysis time is 8 h,enzymolysis temperature 60 ?,the concentration of trypsin 0.12%.2 The separation and purification of active peptideUsing ultrafiltration,Sephadex G-25 and high performance liquid chromatography(HPLC)method for the enzymolysis liquid‘s separation and purification.Text each components' influence to mouse macrophage phagocytizing ability as the indicators of the activity of the peptides.After ultrafiltration,Oratosquilla oratoria enzymolysis liquid is divided into four components OP1,OP2,OP3 and OP4,and the highest immune activity component is OP3.OP3 can promotes RAW264.7's cell activity to 163% when it's concentration is 5mg/mL.Will OP3 further purified by Sephadex G-25 receive components OPS.Collecting OPS and separating it by HPLC,then get four components of HOP1,HOP2,HOP3 andHOP4.Text components' influence to mouse macrophage phagocytizing ability respectively,and the highest activity component is HOP1.3 Peptide's immune activityThrough the establishment of three immune models,study OPS effecting on mouse macrophage effect on cell factor separately,regulating ability under the condition of the inflammatory response,the protection of RAW264.7 under inflammatory response.Models use mouse macrophage induced by LPS as positive control group,and set up the blank control group.Results show that the OPS role within the cells to a certain extent to promote cells release inflammatory factor separately,but can decrease RAW264.7 release inflammatory factors induced by LPS,such as iNOS,NO,TNF-?,while it can increase the production of IL-10 which is suppression of inflammation factor,but the releasing of inflammatory factor effect,IL-6 is not obvious.4 Immune active peptide capsule preparationBased on the above research,making preliminary exploration of immune active peptide ‘s hard capsule preparation.Using medicinal starch as material for three batches of samples' preparation.According to the 2010 edition of pharmacopoeia appendix,the requirements of the relevant quality standards,the paper makes the preliminary quality standards of Oratosquilla oratoria immune active peptide capsule.Then the result of texts such as moisture content,the difference of the load,the disintegration time limit,and microbial limit inspection project,shows that the preparation technology is reasonable,for Oratosquilla oratoria immune active peptide capsule production laid the foundation of industrialization.