The Comparative Study On Protein From Differences Original Bird's Nest

Objective:The main research of bird's nest is its pharmacological effects and chemical components.The bird's nests contain 50%protein.But the protein research has not yet been reported for bird's nest.Generally,people accepted that imports of bird's nest was good.Especially from Vietnam.But,the difference between different sources of bird's nest were not reported.And the protein research developed continuously in recent years.The application of these methods did not used in the bird nest.Therefore,this research was based on the previous studies.The bird's nest water soluble protein was the research object and to establish the determination method of protein content in the bird's nest,optimization of the extraction of total protein in bird's nest,compare the protein of different bird's nests,and explore the function of the protein function.Methods:1.On the basis of single factor experiment optimize soak time,dosage of solvent and ultrasonic time three influence factors in protein of bird's nest.2.Using the BCA method to determine the protein content in the bird's nest.3.Using sds-page,capillary high performance liquid chromatography(HPLC),ESI mass spectrometry to detect different kinds of bird's nest protein,and comparing the water-soluble protein molecular weight and its hydrolysis polypeptide differences in red and white bird's nest.the similarities and differences between protein were determined through the analysis of the database.4.Using sds-page,capillary high performance liquid chromatography(HPLC),ESI mass spectrometry to detect different kinds of bird's nest protein,and comparing the water-soluble protein molecular weight and its hydrolysis polypeptide differences in different genotypes of bird's nest.5.Using the Label-free quantitative method to analysis the protein content in the bird's nest.Results:1.The water soluble protein extraction conditions of extracting time 4 h,material liquid than 1:6 00,ultrasonic time 3.5 h.2.In the range of 25-1000?g/mL,there is a good linear relations-hip(r=0.9999);The average recovery is 99.58%,RSD is 2.17%.The regression equation:y = 0.0007 + 0.0797 x.3.According to the results of the mass spectrometry Basepeak figure:red and white bird's nest hydrolytic polypeptide had good consistency.Using Mascot 2.2 software to download the uniprot of poultry database,it retrievaled and evaluatived 73 kinds of proteins in white,identification of 51 proteins in red,red and white bird's nest had 30 kinds of the same protein.4.According to the results of the mass spectrometry Basepeak figure:the different kinds of hydrolytic polypeptide had certain consistency.Using Mascot 2.2 software to download the uniprot of poultry database,it appraisalled 71 kinds of proteins.Identification of 41 proteins in Huaiji,identification of 54 proteins in Vietnam,47 proteins in Indonesia,28 proteins in feather,The same of proteins had 20 kinds in four kinds of bird's nest.Conclusions:1.In this study,from the protein extraction conditions and protein content,we can ultimately determine the optimal extraction conditions,the water-soluble protein content was measured by BCA;the results show that the extraction conditions is feasible,rapid and accurate.The determination for protein extraction can provide the basis for establishing quality standard.2.Study on comparatived the difference of protein in red and white bird's nest found differences.Although in the spectrum have a higher degree of similarity,but protein identification results showed that red and white bird's nest has significant differences.So they could provide variety identification theoretical basis.3.Study on comparatived the difference of protein in different genotypes of bird's nest found significant differences in protein.The same of protein in different genotypes of bird's nest can be used as quality control of the product characteristics.It provides valuable reference for the establishment of bird's nest quality standards.