Extraction Of Zanthoxylum Seed Protein And Identification And Analysis Of Simulated Digestion Products In Vitro

As the main by-products in the production of Zanthoxylum bungeanum industry,Zanthoxylum bungeanum seeds are rich in nutrients such as melanin,fat and protein,but not be fully utilized.In this study,the Zanthoxylum bungeanum seeds of Hanyuan Gongjiao were used as raw materials,and the following studies were carried out:1.The basic components of Zanthoxylum bungeanum seed were determined.The crude protein content of Zanthoxylum bungeanum seeds is(11.49±0.27)%,and gluten,globulin,gliadin,albumin and other insoluble proteins account for 63.22%,15.44%,2.28%,2.74% and16.30% respectively.2.Alkali-soluble acid precipitation method and salt-soluble method were compared to extract Zanthoxylum bungeanum seed protein.The results showed that both the extraction rate and protein abundance of alkali-soluble acid precipitation method are superior than those of salt-soluble method.Therefore,the alkali-soluble acid precipitation method is used to extract Zanthoxylum bungeanum seed protein.Single factor-based orthogonal method is used to explore the optimal process.The factors affecting the yield of Zanthoxylum bungeanum seed protein were ranked as: alkali dissolution temperature(?)> extraction time(min)>material-liquid ratio(g/m L)> pH.The optimal process were as follow: extraction temperature40 ?,extraction time 40 min,material-liquid ratio 1:20(w/v),pH 10.Under above conditions,the yield of Zanthoxylum bungeanum seed protein reached 33.40%.3.The gastric and intestinal digestion of Zanthoxylum bungeanum seed protein was simulated,and Orbitrap mass spectrometry(Orbitrap)was used to analyze the characteristics of gastrointestinal digestion products.The results showed that the molecular weight of the digestion products is mainly concentrated within 1200 Da,and the strongest signal response is373.25 Da.The results of a series of antioxidant assays showed that the antioxidant activity of digestion products increased continuously over digestion process;the intestinal digestion products showed more antioxidant activity than the stomach digestion products.The antioxidant activity of the final digested product is below VC.4.Nano LC-ESI-MS/MS technology and 6 bioinformatics databases including SATPdb,APD3,Stra Pep,HIPdb,BIOPEP-UWM and AVPdb were combined to identify and predict all peptides in the final product of gastrointestinal simulated digestion of Zanthoxylum bungeanum seed protein.66 suspected bioactive peptides were obtained,51 with antibacterial activity,1 with nerve blocking effect,6 with growth factor-like function,3 with anticancer activity,4 with antiviral activity,and 1 with drug delivery activity.