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首页> 外文期刊>Applied Microbiology >Prolyl Aminopeptidase from Streptomyces thermoluteus subsp. fuscus Strain NBRC14270 and Synthesis of Proline-Containing Peptides by Its S144C Variant
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Prolyl Aminopeptidase from Streptomyces thermoluteus subsp. fuscus Strain NBRC14270 and Synthesis of Proline-Containing Peptides by Its S144C Variant

机译：来自热链霉菌亚种的脯氨酰氨肽酶。品红菌株NBRC14270及其S144C变体合成脯氨酸肽

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摘要

We specifically examined an exopeptidase, prolyl aminopeptidase (PAP), as a target for synthesis of proline-containing peptides. A PAP from Streptomyces thermoluteus subsp. fuscus NBRC14270 ({"type":"entrez-protein","attrs":{"text":"PAP14270","term_id":"1238921194","term_text":"PAP14270"}}PAP14270) was obtained using sequence-based screening. From {"type":"entrez-protein","attrs":{"text":"PAP14270","term_id":"1238921194","term_text":"PAP14270"}}PAP14270, 144Ser was replaced by Cys (scPAP14270) to give aminolysis activity. In contrast to wild-type {"type":"entrez-protein","attrs":{"text":"PAP14270","term_id":"1238921194","term_text":"PAP14270"}}PAP14270, scPAP14270 produced a polymer of proline benzyl ester and cyclo[Pro-Pro]. The product mass was confirmed using liquid chromatography-mass spectrometry (LC/MS). Several factors affecting the reaction, such as the pH, concentration of the substrate, and reaction time, were measured to determine their effects. Furthermore, a correlation was found between substrate specificity in proline peptide synthesis and the log D value of acyl acceptors in aminolysis catalyzed by scPAP14270. Results showed that dipeptide synthesis proceeded in a weakly acidic environment and that cyclization and polymerization occurred under alkaline conditions. Furthermore, results suggest that almost all amino acid esters whose log D value is greater than 0, except hydroxyproline benzyl ester (Hyp-OBzl), can be recognized as acyl acceptors. These findings support the use of PAPs as a tool for production of physiologically active proline peptides.
机译：我们专门检查了一种外肽酶，脯氨酰氨肽酶（PAP），作为合成含有脯氨酸的肽的靶标。来自热链霉菌亚种的PAP。使用序列获得了番红花NBRC14270（{“ type”：“ entrez-protein”，“ attrs”：{“ text”：“ PAP14270”，“ term_id”：“ 1238921194”，“ term_text”：“ PAP14270”}} PAP14270）的筛选。从{“ type”：“ entrez-protein”，“ attrs”：{“ text”：“ PAP14270”，“ term_id”：“ 1238921194”，“ term_text”：“ PAP14270”}} PAP14270，144Ser被Cys（ scPAP14270）具有氨解活性。与野生型{“ type”：“ entrez-protein”，“ attrs”：{“ text”：“ PAP14270”，“ term_id”：“ 1238921194”，“ term_text”：“ PAP14270”}} PAP14270，scPAP14270制得脯氨酸苄酯和环[Pro-Pro]的聚合物。使用液相色谱-质谱法（LC / MS）确认产物质量。测量影响反应的几个因素，例如pH，底物浓度和反应时间，以确定它们的作用。此外，发现脯氨酸肽合成中的底物特异性与scPAP14270催化的氨解中酰基受体的log D值之间存在相关性。结果表明，二肽的合成在弱酸性环境中进行，环化和聚合在碱性条件下发生。此外，结果表明，除了羟脯氨酸苄基酯（Hyp-OBzl）外，几乎所有log D值均大于0的氨基酸酯都可以被识别为酰基受体。这些发现支持将PAP用作生产生理活性脯氨酸肽的工具。

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