The Aminolysis Reaction of
            <i>Streptomyces</i>
            S9 Aminopeptidase Promotes the Synthesis of Diverse Prolyl Dipeptides

Arima, Jiro; Morimoto, Masazumi; Usuki, Hirokazu; Mori, Nobuhiro; Hatanaka, Tadashi

doi:10.1128/aem.00577-10

Cited by 13 publications

(6 citation statements)

References 14 publications

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“…27,28 Aminopeptidases have also been exploited for peptide synthesis under appropriate conditions, and high conversion ratios were achieved. 29,30 The peptides synthesized in the hydrolysis system might be rather few but still could be detected by UPLC-MS/MS. As this study is not able to evaluate the concentration of every peptide, the influence of BLAM on the content of peptides needs further exploration.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

Evaluation of the Hydrolysis Specificity of an Aminopeptidase from Bacillus licheniformis SWJS33 Using Synthetic Peptides and Soybean Protein Isolate

Lei

Zhao

Lin

et al. 2016

J. Agric. Food Chem.

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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.

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Section: ■ Results and Discussionmentioning

confidence: 99%

Evaluation of the Hydrolysis Specificity of an Aminopeptidase from Bacillus licheniformis SWJS33 Using Synthetic Peptides and Soybean Protein Isolate

Lei

Zhao

Lin

et al. 2016

J. Agric. Food Chem.

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“…These peptidases showed the activity only toward D-amino acid residues. Very recently, we have demonstrated that S9 aminopeptidase from S. thermocyaneoviolaceus NBRC 14271 was also catalyzed, and the aminolysis reaction was similar to that by 14271 X-PDAP [19]. The reaction was consisted of L-Pro-OBzl ester as an acyl donor and L-or D-amino acid esters as acyl acceptors, and produced L-Pro-Damino acid esters.…”

Section: Aminolysis Reactionmentioning

confidence: 87%

Extracellular Production and Characterization of Streptomyces X-prolyl Dipeptidyl Aminopeptidase

Hatanaka¹,

Yamasato²,

Arima

et al. 2011

Appl Biochem Biotechnol

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X-prolyl dipeptidyl aminopeptidases (X-PDAPs) are useful in various food industries. In this study, we performed sequence-based screening to obtain a stable X-PDAP enzyme from thermophilic Streptomyces strains. We found three genes that encoded X-PDAP from Streptomyces thermoluteus subsp. fuscus NBRC 14270 (14270 X-PDAP), Streptomyces thermocyaneoviolaceus NBRC 14271 (14271 X-PDAP), and Streptomyces thermocoerulescens NBRC 14273, which were subsequently cloned and sequenced. The deduced amino acid sequences of these genes showed high similarity, with ~80% identity with each other. The isolated X-PDAPs and an X-PDAP from Streptomyces coelicolor were expressed in Streptomyces lividans using a hyperexpression vector: pTONA5a. Among these genes, only 14270 and 14271 X-PDAPs caused overexpression and extracellular production without artificial signal peptides. We also characterized the biochemical properties of purified 14271 X-PDAP. In addition, we found that, in peptide synthesis via an aminolysis reaction, this enzyme recognized D-amino acid derivatives as acyl acceptors, similar to L-amino acid derivatives.

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“…In the presence of relatively high concentrations of primary amine, which are not achieved under physiological conditions, nucleophilic attack by an amino group instead of a water molecule forms an amide bond ( Fig. 1) [6][7][8][9][10][11]. In this reaction, termed aminolysis, the primary amine acts as an acyl acceptor.…”

Section: Introductionmentioning

confidence: 99%

Crystal structure of d‐stereospecific amidohydrolase from Streptomyces sp. 82F2 – insight into the structural factors for substrate specificity

et al. 2015

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D-Stereospecific amidohydrolase (DAH) from Streptomyces sp. 82F2, which catalyzes amide bond formation from D-aminoacyl esters and L-amino acids (aminolysis), can be used to synthesize short peptides with a DL-configuration. We found that DAH can use 1,8-diaminooctane and other amino compounds as acyl acceptors in the aminolysis reaction. Low concentrations of 1,8-diaminooctane inhibited acyl-DAH intermediate formation. By contrast, excess 1,8-diaminooctane promoted aminolysis by DAH, producing D-Phe-1,8-diaminooctane via nucleophilic attack of the diamine on enzyme-bound D-Phe. To clarify the mechanism of substrate specificity and amide bond formation by DAH, the crystal structure of the enzyme that binds 1,8-diaminooctane was determined at a resolution of 1.49A. Comparison of the DAH crystal structure with those of other members of the S12 peptidase family indicated that the substrate specificity of DAH arises from its active site structure. The 1,8-diaminooctane molecule binds at the entrance of the active site pocket. The electrkon density map showed that another potential 1,8-diaminooctane binding site, probably with lower affinity, is present close to the active site. The enzyme kinetics and structural comparisons suggest that the location of enzyme-bound diamine can explain the inhibition of the acyl-enzyme intermediate formation, although the bound diamine is too far from the active site for aminolysis. Despite difficulty in locating the diamine binding site for aminolysis definitively, we propose that the excess diamine also binds at or near the second binding site to attack the acyl-enzyme intermediate during aminolysis.

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The Aminolysis Reaction of Streptomyces S9 Aminopeptidase Promotes the Synthesis of Diverse Prolyl Dipeptides

Cited by 13 publications

References 14 publications

Evaluation of the Hydrolysis Specificity of an Aminopeptidase from Bacillus licheniformis SWJS33 Using Synthetic Peptides and Soybean Protein Isolate

Evaluation of the Hydrolysis Specificity of an Aminopeptidase from Bacillus licheniformis SWJS33 Using Synthetic Peptides and Soybean Protein Isolate

Extracellular Production and Characterization of Streptomyces X-prolyl Dipeptidyl Aminopeptidase

Crystal structure of d‐stereospecific amidohydrolase from Streptomyces sp. 82F2 – insight into the structural factors for substrate specificity

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