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Improvement in organophosphorus hydrolase activity of cell surface-engineered yeast strain using Flo1p anchor system

Organophosphorus hydrolase (OPH) hydrolyzes organophosphorus esters. We constructed the yeast-displayed OPH using Flo1p anchor system. In this system, the N-terminal region of the protein was fused to Flo1p and the fusion protein was displayed on the cell surface. Hydrolytic reactions with paraoxon...

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Autores principales: Fukuda, Takeshi, Tsuchiyama, Kouta, Makishima, Hirokazu, Takayama, Katsumi, Mulchandani, Ashok, Kuroda, Kouichi, Ueda, Mitsuyoshi, Suye, Shin-ichiro
Formato: Texto
Lenguaje:English
Publicado: Springer Netherlands 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2852028/
https://www.ncbi.nlm.nih.gov/pubmed/20111980
http://dx.doi.org/10.1007/s10529-010-0204-1
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author Fukuda, Takeshi
Tsuchiyama, Kouta
Makishima, Hirokazu
Takayama, Katsumi
Mulchandani, Ashok
Kuroda, Kouichi
Ueda, Mitsuyoshi
Suye, Shin-ichiro
author_facet Fukuda, Takeshi
Tsuchiyama, Kouta
Makishima, Hirokazu
Takayama, Katsumi
Mulchandani, Ashok
Kuroda, Kouichi
Ueda, Mitsuyoshi
Suye, Shin-ichiro
author_sort Fukuda, Takeshi
collection PubMed
description Organophosphorus hydrolase (OPH) hydrolyzes organophosphorus esters. We constructed the yeast-displayed OPH using Flo1p anchor system. In this system, the N-terminal region of the protein was fused to Flo1p and the fusion protein was displayed on the cell surface. Hydrolytic reactions with paraoxon were carried out during 24 h of incubation of OPH-displaying cells at 30°C. p-Nitrophenol produced in the reaction mixture was detected by HPLC. The strain with highest activity showed 8-fold greater OPH activity compared with cells engineered using glycosylphosphatidylinositol anchor system, and showed 20-fold greater activity than Escherichia coli using the ice nucleation protein anchor system. These results indicate that Flo1p anchor system is suitable for display of OPH in the cell surface-expression systems.
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spelling pubmed-28520282010-04-16 Improvement in organophosphorus hydrolase activity of cell surface-engineered yeast strain using Flo1p anchor system Fukuda, Takeshi Tsuchiyama, Kouta Makishima, Hirokazu Takayama, Katsumi Mulchandani, Ashok Kuroda, Kouichi Ueda, Mitsuyoshi Suye, Shin-ichiro Biotechnol Lett Original Research Paper Organophosphorus hydrolase (OPH) hydrolyzes organophosphorus esters. We constructed the yeast-displayed OPH using Flo1p anchor system. In this system, the N-terminal region of the protein was fused to Flo1p and the fusion protein was displayed on the cell surface. Hydrolytic reactions with paraoxon were carried out during 24 h of incubation of OPH-displaying cells at 30°C. p-Nitrophenol produced in the reaction mixture was detected by HPLC. The strain with highest activity showed 8-fold greater OPH activity compared with cells engineered using glycosylphosphatidylinositol anchor system, and showed 20-fold greater activity than Escherichia coli using the ice nucleation protein anchor system. These results indicate that Flo1p anchor system is suitable for display of OPH in the cell surface-expression systems. Springer Netherlands 2010-01-29 2010 /pmc/articles/PMC2852028/ /pubmed/20111980 http://dx.doi.org/10.1007/s10529-010-0204-1 Text en © The Author(s) 2010 https://creativecommons.org/licenses/by-nc/4.0/ This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and source are credited.
spellingShingle Original Research Paper
Fukuda, Takeshi
Tsuchiyama, Kouta
Makishima, Hirokazu
Takayama, Katsumi
Mulchandani, Ashok
Kuroda, Kouichi
Ueda, Mitsuyoshi
Suye, Shin-ichiro
Improvement in organophosphorus hydrolase activity of cell surface-engineered yeast strain using Flo1p anchor system
title Improvement in organophosphorus hydrolase activity of cell surface-engineered yeast strain using Flo1p anchor system
title_full Improvement in organophosphorus hydrolase activity of cell surface-engineered yeast strain using Flo1p anchor system
title_fullStr Improvement in organophosphorus hydrolase activity of cell surface-engineered yeast strain using Flo1p anchor system
title_full_unstemmed Improvement in organophosphorus hydrolase activity of cell surface-engineered yeast strain using Flo1p anchor system
title_short Improvement in organophosphorus hydrolase activity of cell surface-engineered yeast strain using Flo1p anchor system
title_sort improvement in organophosphorus hydrolase activity of cell surface-engineered yeast strain using flo1p anchor system
topic Original Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2852028/
https://www.ncbi.nlm.nih.gov/pubmed/20111980
http://dx.doi.org/10.1007/s10529-010-0204-1
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