title: Evaluation of the Hydrolysis Specificity of an Aminopeptidase from Bacillus licheniformis SWJS33 Using Synthetic Peptides and Soybean Protein Isolate.
abstract: The substrate specificity of aminopeptidases has often been determined against aminoacyl-p-nitroanilide; thus, its specificity toward synthetic peptides and complex substrates remained unclear. The hydrolysis specificity of an aminopeptidase from Bacillus licheniformis SWJS33 (BLAM) was evaluated using a series of synthetic peptides and soybean protein isolate. The aminopeptidase showed high specificity for dipeptides with Leu, Val, Ala, Gly, and Phe at the N-terminus, and the specificity was significantly affected by the nature of the penultimate residue. In the hydrolysis of soy protein isolate, BLAM preferred peptides with Leu, Glu, Gly, and Ala at the N-terminus by free amino acid analysis and preferred peptides with Leu, Ala, Ser, Trp, and Tyr at the N-terminus by UPLC-MS/MS. The introduction of complex substrates provides a deeper understanding of the aminopeptidase's specificity, which can instruct the application of the enzyme in protein hydrolysis.
authors.affiliation: School of Food Science and Engineering, South China University of Technology , Guangzhou, China 510640.
authors.forename: Fenfen
authors.initials: F
authors.lastname: Lei
authors.affiliation: School of Food Science and Engineering, South China University of Technology , Guangzhou, China 510640.
authors.forename: Qiangzhong
authors.initials: Q
authors.lastname: Zhao
authors.affiliation: School of Food Science and Engineering, South China University of Technology , Guangzhou, China 510640.
authors.forename: Lianzhu
authors.initials: L
authors.lastname: Lin
authors.affiliation: Beijing Key Laboratory of Flavor Chemistry, Beijing Technology and Business University , Beijing, China 100048.
authors.forename: Baoguo
authors.initials: B
authors.lastname: Sun
authors.affiliation: School of Food Science and Engineering, South China University of Technology , Guangzhou, China 510640.|Advanced Innovation Center for Food Nutrition and Human Health, Beijing Technology and Business University , Beijing, China 100048.
authors.forename: Mouming
authors.identifier: http://orcid.org/0000-0003-0221-3838
authors.initials: M
authors.lastname: Zhao
mesh_terms: D000595:Amino Acid Sequence; D000626:Aminopeptidases / Q000737:chemistry* / Q000378:metabolism; D000069976:Bacillus licheniformis / Q000737:chemistry / Q000201:enzymology*; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D006868:Hydrolysis; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D030262:Soybean Proteins / Q000737:chemistry* / Q000378:metabolism; D013379:Substrate Specificity; D053719:Tandem Mass Spectrometry
chemical_list: D001426:Bacterial Proteins; D010455:Peptides; D030262:Soybean Proteins; D000626:Aminopeptidases
keywords: UPLC-MS/MS; aminopeptidase; free amino acid; hydrolysis specificity; synthetic peptides