A mutant phospholipase D with enhanced thermostability from Streptomyces sp.

Tadashi Hatanaka; Tomofumi Negishi; Koichi Mori

doi:10.1016/j.bbapap.2003.09.013

A mutant phospholipase D with enhanced thermostability from Streptomyces sp.

Tadashi Hatanaka, Tomofumi Negishi, Koichi Mori

Research output: Contribution to journal › Article › peer-review

20 Citations (Scopus)

Abstract

To investigate the contribution of amino acid residues to the thermostability of phospholipase D (PLD), a chimeric form of two Streptomyces PLDs (thermolabile K1PLD and thermostable TH-2PLD) was constructed. K/T/KPLD, in which residues 329-441 of K1PLD were recombined with the homologous region of TH-2PLD, showed a thermostability midway between those of K1PLD and TH-2PLD. By comparing the primary structures of Streptomyces PLDs, the seven candidates of thermostability-related amino acid residues of K1PLD were identified. The K1E346DPLD mutant, in which Glu346 of K1PLD was substituted with Asp by site-directed mutagenesis, exhibited enhanced thermostability, which was almost the same as that of TH-2PLD.

Original language	English
Pages (from-to)	75-82
Number of pages	8
Journal	Biochimica et Biophysica Acta - Proteins and Proteomics
Volume	1696
Issue number	1
DOIs	https://doi.org/10.1016/j.bbapap.2003.09.013
Publication status	Published - Jan 14 2004
Externally published	Yes

Keywords

Biocatalyst
Phospholipase D
Streptomyces
Thermostability

ASJC Scopus subject areas

Analytical Chemistry
Biophysics
Biochemistry
Molecular Biology

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10.1016/j.bbapap.2003.09.013

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abstract = "To investigate the contribution of amino acid residues to the thermostability of phospholipase D (PLD), a chimeric form of two Streptomyces PLDs (thermolabile K1PLD and thermostable TH-2PLD) was constructed. K/T/KPLD, in which residues 329-441 of K1PLD were recombined with the homologous region of TH-2PLD, showed a thermostability midway between those of K1PLD and TH-2PLD. By comparing the primary structures of Streptomyces PLDs, the seven candidates of thermostability-related amino acid residues of K1PLD were identified. The K1E346DPLD mutant, in which Glu346 of K1PLD was substituted with Asp by site-directed mutagenesis, exhibited enhanced thermostability, which was almost the same as that of TH-2PLD.",

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AU - Hatanaka, Tadashi

AU - Negishi, Tomofumi

AU - Mori, Koichi

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AB - To investigate the contribution of amino acid residues to the thermostability of phospholipase D (PLD), a chimeric form of two Streptomyces PLDs (thermolabile K1PLD and thermostable TH-2PLD) was constructed. K/T/KPLD, in which residues 329-441 of K1PLD were recombined with the homologous region of TH-2PLD, showed a thermostability midway between those of K1PLD and TH-2PLD. By comparing the primary structures of Streptomyces PLDs, the seven candidates of thermostability-related amino acid residues of K1PLD were identified. The K1E346DPLD mutant, in which Glu346 of K1PLD was substituted with Asp by site-directed mutagenesis, exhibited enhanced thermostability, which was almost the same as that of TH-2PLD.

KW - Biocatalyst

KW - Phospholipase D

KW - Streptomyces

KW - Thermostability

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A mutant phospholipase D with enhanced thermostability from Streptomyces sp.

Abstract

Keywords

ASJC Scopus subject areas

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