Structure-Based Modification of D-Alanine-D-Alanine Ligase from Thermotoga maritima ATCC 43589 for Depsipeptide Synthesis

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概要

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• Depsipeptides are peptide-like polymers consisting of amino acids and hydroxy acids, and are expected to be new functional materials for drug-delivery systems and polymer science. In our previous study, D-alanyl-D-lactate, a type of depsipeptide, was enzymatically synthesized using D-alanine-D-alanine ligase from Thermotoga maritima ATCC 43589 (TmDdl) by Y207F substitution. Thereafter, in this study, further mutagenesis was introduced, based on structural comparison between TmDdl and a well-characterized D-alanine-D-alanine ligase from Escherichia coli. The S137A/Y207F mutant showed higher D-alanyl-D-lactate and lower D-alanyl-D-alanine synthesizing activity than the Y207F mutant. This suggests that substitution at the S137 residue contributes to product selectivity. Saturated mutagenesis on S137 revealed that the S137G/Y207F mutant showed the highest D-alanyl-D-lactate synthesizing activity. Moreover, the mutant showed broad substrate specificity toward D-amino acid and recognized D-lactate and D,L-isoserine as substrates. On the basis of these characteristics, various depsipeptides can be produced using S137G/Y207F-replaced TmDdl.

• 2011-04-23

著者

• Kino K

  Department Of Applied Chemistry School Of Science And Engineering Waseda University

• Kino Kuniki

  Department Of Applied Chemistry Faculty Of Science And Engineering Waseda University

• Kino Kuniki

  Department Of Applied Chemistry School Of Science And Engineering Waseda University

• Kino Kuniki

  Dep. Of Applied Chemistry Fac. Of Sci. And Engineering Waseda Univ. 3-4-1 Ohkubo Shinjuku-ku Tokyo 1

• Satake Ryoko

  Department Of Applied Chemistry Faculty Of Science And Engineering Waseda University

• Sato Masaru

  Department Of Applied Chemistry School Of Advanced Science And Engineering Faculty Of Science And En

• NAKAGAWA Tomoki

  Department of Applied Chemistry, School of Advanced Science and Engineering, Faculty of Science and

• Nakagawa Tomoki

  Department Of Applied Chemistry School Of Advanced Science And Engineering Faculty Of Science And En

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