Keywords

Imitation Sea Cucumber, Haitian Sweet Potato, Response Surface, Peptide Isolation And Purification, N-Terminal Sequencing

Abstract

In order to fully develop and utilize sea cucumber resources, optimize the enzymatic hydrolysis process of sea cucumber body wall, and identify sea cucumber peptides, this paper uses the degree of hydrolysis as an indicator to design a response surface experiment with 3 factors (time, temperature, and enzyme amount) and 3 levels, and obtain the results of sea cucumber hydrolysis. The optimal conditions are: the body wall of Apostichopus japonicus has a hydrolysis degree of 83.39% when the amount of enzyme is 1.97%, the temperature is 55.7¡ãC, and the hydrolysis time is 136.8 minutes; the body wall of Acaudina leucoprocta is hydrolyzed by 1.97% enzyme. Under the conditions of 1.70%, temperature 55.4¡æ and hydrolysis 115.6min, the degree of hydrolysis is 63.68%. Four polypeptides were then separated and identified from the hydrolyzate by using ultrafiltration membrane, reversed-phase high-performance liquid chromatography (RP-HPLC), matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF) technology, and protein sequencers. . Among them, the N-terminal sequence identified the amino acid residue sequences of S1 and S2 of sea cucumber-like polypeptides as Gly-Pro-Val-Gly-Ala-Ser-Gly-Pro-Gln-Gly-Pro-Gln-Gly-Pro-Gln respectively. – Gly-Leu-Ser-Ala-Leu and Trp-Pro-Pro-Gly-Asn-Ser-Gly-Ile-Gln-Gly. The amino acid residue sequences of Haitian melon polypeptides A1 and A2 are Gly-Ala-Asn-Gly-Asn and Trp-Leu-Pro-Gly-Asp-Thr-Gly-Pro-Gln-Gly-Val-Thr-Gly-Pro- respectively. Val-Gly-Pro-Ala-Gly.

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Original research done by Li Yanyan, Dai Juan, Hu Lingping, Jiang Zhenzhou, Shang Jing, Zhang Luyong

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