Recombinant production and evaluation of an antibacterial l -amino acid oxidase derived from flounder Platichthys stellatus

Verfasser / Beitragende:
[Kosuke Kasai, Kenro Hashiguchi, Hiroki Takahashi, Ayano Kasai, Sadanori Takeda, Manabu Nakano, Takashi Ishikawa, Toshiya Nakamura, Tomisato Miura]
Ort, Verlag, Jahr:
2015
Enthalten in:
Applied Microbiology and Biotechnology, 99/16(2015-08-01), 6693-6703
Format:
Artikel (online)
ID: 605498369
LEADER caa a22 4500
001 605498369
003 CHVBK
005 20210128100545.0
007 cr unu---uuuuu
008 210128e20150801xx s 000 0 eng
024 7 0 |a 10.1007/s00253-015-6428-1  |2 doi 
035 |a (NATIONALLICENCE)springer-10.1007/s00253-015-6428-1 
245 0 0 |a Recombinant production and evaluation of an antibacterial l -amino acid oxidase derived from flounder Platichthys stellatus  |h [Elektronische Daten]  |c [Kosuke Kasai, Kenro Hashiguchi, Hiroki Takahashi, Ayano Kasai, Sadanori Takeda, Manabu Nakano, Takashi Ishikawa, Toshiya Nakamura, Tomisato Miura] 
520 3 |a Fish produce mucus substances as a defensive outer barrier against several bacterial infections. We have recently identified an antibacterial l-amino acid oxidase (psLAAO1) in the mucus layer of the flounder Platichthys stellate. In this study, the antibacterial protein psLAAO1 was expressed as a secretory bioactive recombinant protein in the methylotrophic yeast Pichia pastoris. The recombinant psLAAO1 inhibited the growth of bacteria to the same levels as native psLAAO1 present in mucus. In particular, Staphylococci and Yersinia were strongly suppressed, showing the highest growth retardation of the 21 species and strains tested. Moreover, Staphylococcus epidermidis was most sensitive to psLAAO1 with a minimum inhibitory concentration (MIC) of 0.078μg/mL, whereas Escherichia coli was essentially resistant to psLAAO1 with a MIC of >10μg/mL. Interestingly, psLAAO1-treated E. coli were found to upregulate the expression of the btuE gene, which encodes glutathione peroxidase (GPx). The biochemical function of GPx is to reduce freehydrogen peroxide and is induced under response to reactive oxygen species (ROS). Thus, E. coli confers resistance to the reduced free hydrogen peroxide produced by psLAAO1 by increasing GPx levels. Furthermore, the growth of Staphylococcus aureus was completely inhibited in the presence of recombinant psLAAO1. The morphology of psLAAO1-treated S. aureus showed cell surface damage, the formation of large aggregates and the cells showed severe deformations. Western blot analysis showed that psLAAO1 binds to the surface of S. aureus. Therefore, psLAAO1 binds to the surface of LAAO-sensitive S. aureus and directs peroxidative activity at the surface of the bacterial membrane. 
540 |a Springer-Verlag Berlin Heidelberg, 2015 
690 7 |a l -Amino acid oxidase  |2 nationallicence 
690 7 |a Antibacterial protein  |2 nationallicence 
690 7 |a Staphylococcus aureus  |2 nationallicence 
690 7 |a Platichthys stellatus  |2 nationallicence 
690 7 |a Glutathione peroxidase  |2 nationallicence 
690 7 |a MRSA  |2 nationallicence 
700 1 |a Kasai  |D Kosuke  |u Department of Pathologic Analysis, Division of Medical Life Sciences, Graduate School of Health Sciences, Hirosaki University, 66-1 Hon-cho, Hirosaki, Aomori, Japan  |4 aut 
700 1 |a Hashiguchi  |D Kenro  |u Department of Clinical Laboratory, Yokohama City University Medical Center, Kanagawa, Japan  |4 aut 
700 1 |a Takahashi  |D Hiroki  |u I'ROM Co., Ltd, Tokyo, Japan  |4 aut 
700 1 |a Kasai  |D Ayano  |u Department of Clinical Laboratory, Hirosaki Municipal Hospital, Aomori, Japan  |4 aut 
700 1 |a Takeda  |D Sadanori  |u Department of Medical Technology, School of Health Sciences, Hirosaki University, Aomori, Japan  |4 aut 
700 1 |a Nakano  |D Manabu  |u Department of Biomedical Sciences, Division of Medical Life Sciences, Graduate School of Health Sciences, Hirosaki University, Aomori, Japan  |4 aut 
700 1 |a Ishikawa  |D Takashi  |u Department of Biomedical Sciences, Division of Medical Life Sciences, Graduate School of Health Sciences, Hirosaki University, Aomori, Japan  |4 aut 
700 1 |a Nakamura  |D Toshiya  |u Department of Biomedical Sciences, Division of Medical Life Sciences, Graduate School of Health Sciences, Hirosaki University, Aomori, Japan  |4 aut 
700 1 |a Miura  |D Tomisato  |u Department of Pathologic Analysis, Division of Medical Life Sciences, Graduate School of Health Sciences, Hirosaki University, 66-1 Hon-cho, Hirosaki, Aomori, Japan  |4 aut 
773 0 |t Applied Microbiology and Biotechnology  |d Springer Berlin Heidelberg  |g 99/16(2015-08-01), 6693-6703  |x 0175-7598  |q 99:16<6693  |1 2015  |2 99  |o 253 
856 4 0 |u https://doi.org/10.1007/s00253-015-6428-1  |q text/html  |z Onlinezugriff via DOI 
898 |a BK010053  |b XK010053  |c XK010000 
900 7 |a Metadata rights reserved  |b Springer special CC-BY-NC licence  |2 nationallicence 
908 |D 1  |a research-article  |2 jats 
949 |B NATIONALLICENCE  |F NATIONALLICENCE  |b NL-springer 
950 |B NATIONALLICENCE  |P 856  |E 40  |u https://doi.org/10.1007/s00253-015-6428-1  |q text/html  |z Onlinezugriff via DOI 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Kasai  |D Kosuke  |u Department of Pathologic Analysis, Division of Medical Life Sciences, Graduate School of Health Sciences, Hirosaki University, 66-1 Hon-cho, Hirosaki, Aomori, Japan  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Hashiguchi  |D Kenro  |u Department of Clinical Laboratory, Yokohama City University Medical Center, Kanagawa, Japan  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Takahashi  |D Hiroki  |u I'ROM Co., Ltd, Tokyo, Japan  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Kasai  |D Ayano  |u Department of Clinical Laboratory, Hirosaki Municipal Hospital, Aomori, Japan  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Takeda  |D Sadanori  |u Department of Medical Technology, School of Health Sciences, Hirosaki University, Aomori, Japan  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Nakano  |D Manabu  |u Department of Biomedical Sciences, Division of Medical Life Sciences, Graduate School of Health Sciences, Hirosaki University, Aomori, Japan  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Ishikawa  |D Takashi  |u Department of Biomedical Sciences, Division of Medical Life Sciences, Graduate School of Health Sciences, Hirosaki University, Aomori, Japan  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Nakamura  |D Toshiya  |u Department of Biomedical Sciences, Division of Medical Life Sciences, Graduate School of Health Sciences, Hirosaki University, Aomori, Japan  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Miura  |D Tomisato  |u Department of Pathologic Analysis, Division of Medical Life Sciences, Graduate School of Health Sciences, Hirosaki University, 66-1 Hon-cho, Hirosaki, Aomori, Japan  |4 aut 
950 |B NATIONALLICENCE  |P 773  |E 0-  |t Applied Microbiology and Biotechnology  |d Springer Berlin Heidelberg  |g 99/16(2015-08-01), 6693-6703  |x 0175-7598  |q 99:16<6693  |1 2015  |2 99  |o 253