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Heterologous expression and activity verification of ornithine decarboxylase from a wild strain of Shewanella xiamenensis

Shewanella xiamenensis is widely found in spoilage fish, shrimp and other seafoods. Under suitable conditions, ornithine can be synthesized into putrescine, which may spoil food or endanger health. Our research used a wild strain of Shewanella xiamenensis isolated from “Yi Lu Xian” salted fish (a sa...

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Autores principales: Liu, Chang, Wang, Guiyuan, Han, Xiangning, Cao, Limin, Wang, Kaiqiang, Lin, Hong, Sui, Jianxin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9808388/
https://www.ncbi.nlm.nih.gov/pubmed/36605515
http://dx.doi.org/10.3389/fmicb.2022.1100889
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author Liu, Chang
Wang, Guiyuan
Han, Xiangning
Cao, Limin
Wang, Kaiqiang
Lin, Hong
Sui, Jianxin
author_facet Liu, Chang
Wang, Guiyuan
Han, Xiangning
Cao, Limin
Wang, Kaiqiang
Lin, Hong
Sui, Jianxin
author_sort Liu, Chang
collection PubMed
description Shewanella xiamenensis is widely found in spoilage fish, shrimp and other seafoods. Under suitable conditions, ornithine can be synthesized into putrescine, which may spoil food or endanger health. Our research used a wild strain of Shewanella xiamenensis isolated from “Yi Lu Xian” salted fish (a salting method for sea bass) as a research object. According to the database of National Center of Biotechnology Information (NCBI), the target ornithine decarboxylase (ODC) gene SpeF was successfully amplified using the wild strain of Shewanella xiamenensis as the template. Sequencing alignment showed that the SpeF of the wild strain had more than 98% homology compared with the standard strain. The amino acid substitution occurred in the residues of 343, 618, 705, and 708 in the wild strain. After optimizing the expression conditions, a heterologous expression system of ODC was constructed to achieve a high yield of expression. The amount of 253.38 mg of ODC per liter of LB broth was finally expressed. High performance liquid chromatography (HPLC) and subsequent ODC activity verification experiments showed that hetero-expressed ODC showed a certain enzyme activity for about 11.91 ± 0.38 U/mg. Our study gives a new way to the development of a low-cost and high-yield strategy to produce ODC, providing experimental materials for further research and elimination of putrescine in food hazards.
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spelling pubmed-98083882023-01-04 Heterologous expression and activity verification of ornithine decarboxylase from a wild strain of Shewanella xiamenensis Liu, Chang Wang, Guiyuan Han, Xiangning Cao, Limin Wang, Kaiqiang Lin, Hong Sui, Jianxin Front Microbiol Microbiology Shewanella xiamenensis is widely found in spoilage fish, shrimp and other seafoods. Under suitable conditions, ornithine can be synthesized into putrescine, which may spoil food or endanger health. Our research used a wild strain of Shewanella xiamenensis isolated from “Yi Lu Xian” salted fish (a salting method for sea bass) as a research object. According to the database of National Center of Biotechnology Information (NCBI), the target ornithine decarboxylase (ODC) gene SpeF was successfully amplified using the wild strain of Shewanella xiamenensis as the template. Sequencing alignment showed that the SpeF of the wild strain had more than 98% homology compared with the standard strain. The amino acid substitution occurred in the residues of 343, 618, 705, and 708 in the wild strain. After optimizing the expression conditions, a heterologous expression system of ODC was constructed to achieve a high yield of expression. The amount of 253.38 mg of ODC per liter of LB broth was finally expressed. High performance liquid chromatography (HPLC) and subsequent ODC activity verification experiments showed that hetero-expressed ODC showed a certain enzyme activity for about 11.91 ± 0.38 U/mg. Our study gives a new way to the development of a low-cost and high-yield strategy to produce ODC, providing experimental materials for further research and elimination of putrescine in food hazards. Frontiers Media S.A. 2022-12-20 /pmc/articles/PMC9808388/ /pubmed/36605515 http://dx.doi.org/10.3389/fmicb.2022.1100889 Text en Copyright © 2022 Liu, Wang, Han, Cao, Wang, Lin and Sui. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Liu, Chang
Wang, Guiyuan
Han, Xiangning
Cao, Limin
Wang, Kaiqiang
Lin, Hong
Sui, Jianxin
Heterologous expression and activity verification of ornithine decarboxylase from a wild strain of Shewanella xiamenensis
title Heterologous expression and activity verification of ornithine decarboxylase from a wild strain of Shewanella xiamenensis
title_full Heterologous expression and activity verification of ornithine decarboxylase from a wild strain of Shewanella xiamenensis
title_fullStr Heterologous expression and activity verification of ornithine decarboxylase from a wild strain of Shewanella xiamenensis
title_full_unstemmed Heterologous expression and activity verification of ornithine decarboxylase from a wild strain of Shewanella xiamenensis
title_short Heterologous expression and activity verification of ornithine decarboxylase from a wild strain of Shewanella xiamenensis
title_sort heterologous expression and activity verification of ornithine decarboxylase from a wild strain of shewanella xiamenensis
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9808388/
https://www.ncbi.nlm.nih.gov/pubmed/36605515
http://dx.doi.org/10.3389/fmicb.2022.1100889
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