Strong Enhancement of Recombinant Cytosine Deaminase Activity in Bifidobacterium longum for Tumor-Targeting Enzyme/Prodrug Therapy

Yoshinori HAMAJI; Minoru FUJIMORI; Takayuki SASAKI; Hitomi MATSUHASHI; Keiichi MATSUI-SEKI; Yuko SHIMATANI-SHIBATA; Yasunobu KANO; Jun AMANO; Shun’ichiro TANIGUCHI

doi:10.1271/bbb.60502

Biochemistry & Molecular Biology Regular Papers

Strong Enhancement of Recombinant Cytosine Deaminase Activity in Bifidobacterium longum for Tumor-Targeting Enzyme/Prodrug Therapy

Yoshinori HAMAJI, Minoru FUJIMORI, Takayuki SASAKI, Hitomi MATSUHASHI, Keiichi MATSUI-SEKI, Yuko SHIMATANI-SHIBATA, Yasunobu KANO, Jun AMANO, Shun’ichiro TANIGUCHI

Author information

Yoshinori HAMAJI
Department of Surgery, Shinshu University School of Medicine
Minoru FUJIMORI
Department of Surgery, Shinshu University School of Medicine
Takayuki SASAKI
Department of Surgery, Shinshu University School of Medicine
Hitomi MATSUHASHI
Department of Surgery, Shinshu University School of Medicine
Keiichi MATSUI-SEKI
Department of Surgery, Shinshu University School of Medicine
Yuko SHIMATANI-SHIBATA
Department of Surgery, Shinshu University School of Medicine
Yasunobu KANO
Department of Molecular Genetics, Institute of Molecular and Cellular Biology for Pharmaceutical Sciences, Kyoto Pharmaceutical University
Jun AMANO
Department of Surgery, Shinshu University School of Medicine
Shun’ichiro TANIGUCHI
Department of Molecular Oncology, Institute on Aging and Adaptation, Shinshu University Graduate School of Medicine

Keywords: Bifidobacterium longum, shuttle-plasmid, cytosine deaminase, enzyme/prodrug strategy, 5-fluorouracil (5-FU)

JOURNAL FREE ACCESS

2007 Volume 71 Issue 4 Pages 874-883

DOI https://doi.org/10.1271/bbb.60502

View "Advance Publication" version (April 7, 2007).

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Abstract

In our previous studies, a strain of the nonpathogenic, anaerobic, intestinal bacterium, Bifidobacterium longum (B. longum), was found to be localized selectively and to proliferate within solid tumors after systemic administration. In addition, B. longum transformed with the shuttle-plasmid encoding the cytosine deaminase (CD) gene expressed active CD, which deaminated the prodrug 5-fluorocytosine (5-FC) to the anticancer agent 5-fluorouracil (5-FU). We also reported antitumor efficacy with the same plasmid in several animal experiments. In this study, we constructed a novel shuttle-plasmid, pAV001-HU-eCD-M968, which included the mutant CD gene with a mutation at the active site to increase the enzymatic activity.
In addition, the plasmid-transformed B. longum produces mutant CD and strongly increased (by 10-fold) its 5-FC to 5-FU enzymatic activity. The use of B. longum harboring the new shuttle-plasmid increases the effectiveness of our enzyme/prodrug strategy.

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