A mutant leucine aminopeptidase from Streptomyces cinnamoneus with enhanced l-aspartyl l-amino acid methyl ester synthetic activity

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• 作者：Jiro Arima (1) arima@muses.tottori-u.ac.jp
  Mirai Kono (1) m-kono@bs.naist.jp
  Manami Kita (1) manammitai@yahoo.co.jp
  Nobuhiro Mori (1) morinobu@muses.tottori-u.ac.jp
• 关键词：Aminopeptidase &#8211 ; l ; aspartyl ; l ; phenylalanine methyl ester &#8211 ; Peptide bond formation &#8211 ; Reverse reaction
• 刊名：Biotechnology Letters
• 出版年：2012
• 出版时间：June 2012
• 年：2012
• 卷：34
• 期：6
• 页码：1093-1099
• 全文大小：349.5 KB
• 参考文献：1. Allain CC, Poon LS, Chan CS, Richmond W, Fu PC (1974) Enzymatic determination of total serum cholesterol. Clin Chem 20:470–475
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  3. Arima J, Uesugi Y, Iwabuchi M, Hatanaka T (2005) Alteration of leucine aminopeptidase from Streptomyces septatus TH-2 to phenylalanine aminopeptidase by site-directed mutagenesis. Appl Environ Microbiol 71:7229–7235
  4. Arima J, Uesugi Y, Uraji M, Iwabuchi M, Hatanaka T (2006a) Dipeptide synthesis by an aminopeptidase from Streptomyces septatus TH-2 and its application to synthesis of biologically active peptides. Appl Environ Microbiol 72:4225–4231
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• 作者单位：1. Department of Agricultural, Biological, and Environmental Sciences, Faculty of Agriculture, Tottori University, Tottori, 680-8553 Japan
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Life Sciences
  Microbiology
  Biotechnology
  Applied Microbiology
  Biochemistry
  
• 出版者：Springer Netherlands
• ISSN：1573-6776

文摘

l-Aspartyl l-amino acid methyl ester was synthesized using a mutant of a thermostable leucine aminopeptidase from Streptomyces cinnamoneus, D198 K SSAP, obtained in previously. A peptide of high-intensity sweetener, l-aspartyl-l-phenylalanine methyl ester, was selected as a model for demonstrating the synthesis of l-aspartyl l-amino acid methyl ester. The hydrolytic activities of D198 K SSAP toward l-aspartyl-l-phenylalanine and its methyl ester were, respectively, 74-fold and fourfold higher than those of wild type. Similarly, the initial rate of the enzyme for l-aspartyl-l-phenylalanine methyl ester synthesis was over fivefold higher than that of wild-type SSAP in 90% methanol (v/v) in a one-pot reaction. Furthermore, other l-aspartyl l-amino acid methyl esters were synthesized efficiently using D198 K SSAP. Results show that the substitution of Asp198 of SSAP with Lys is effective for synthesizing l-aspartyl l-amino acid methyl ester.