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Boosting Auto-Induction of Recombinant Proteins in Escherichia coli with Glucose and Lactose Additives
BACKGROUND: Auto-induction is a convenient way to produce recombinant proteins without inducer addition using lac operon-controlled Escherichia coli expression systems. Auto-induction can occur unintentionally using a complex culture medium prepared by mixing culture substrates. The differences in c...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Bentham Science Publishers
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8811614/ https://www.ncbi.nlm.nih.gov/pubmed/34353248 http://dx.doi.org/10.2174/0929866528666210805120715 |
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author | Tahara, Nariyasu Tachibana, Itaru Takeo, Kazuyo Yamashita, Shinji Shimada, Atsuhiro Hashimoto, Misuzu Ohno, Satoshi Yokogawa, Takashi Nakagawa, Tsutomu Suzuki, Fumiaki Ebihara, Akio |
author_facet | Tahara, Nariyasu Tachibana, Itaru Takeo, Kazuyo Yamashita, Shinji Shimada, Atsuhiro Hashimoto, Misuzu Ohno, Satoshi Yokogawa, Takashi Nakagawa, Tsutomu Suzuki, Fumiaki Ebihara, Akio |
author_sort | Tahara, Nariyasu |
collection | PubMed |
description | BACKGROUND: Auto-induction is a convenient way to produce recombinant proteins without inducer addition using lac operon-controlled Escherichia coli expression systems. Auto-induction can occur unintentionally using a complex culture medium prepared by mixing culture substrates. The differences in culture substrates sometimes lead to variations in the induction level. OBJECTIVES: In this study, we investigated the feasibility of using glucose and lactose as boosters of auto-induction with a complex culture medium. METHODS: First, auto-induction levels were assessed by quantifying recombinant GFPuv expression under the control of the T7 lac promoter. Effectiveness of the additive-containing medium was examined using ovine angiotensinogen (tac promoter-based expression) and Thermus thermophilus manganese-catalase (T7 lac promoter-based expression). RESULTS: Auto-induced GFPuv expression was observed with the enzymatic protein digest Polypepton, but not with another digest tryptone. Regardless of the type of protein digest, supplementing Terrific Broth medium with glucose (at a final concentration of 2.9 g/L) and lactose (at a final concentration of 7.6 g/L) was successful in obtaining an induction level similar to that achieved with a commercially available auto-induction medium. The two recombinant proteins were produced in milligram quantity of purified protein per liter of culture. CONCLUSION: The medium composition shown in this study would be practically useful for attaining reliable auto-induction for E. coli-based recombinant protein production. |
format | Online Article Text |
id | pubmed-8811614 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Bentham Science Publishers |
record_format | MEDLINE/PubMed |
spelling | pubmed-88116142022-03-02 Boosting Auto-Induction of Recombinant Proteins in Escherichia coli with Glucose and Lactose Additives Tahara, Nariyasu Tachibana, Itaru Takeo, Kazuyo Yamashita, Shinji Shimada, Atsuhiro Hashimoto, Misuzu Ohno, Satoshi Yokogawa, Takashi Nakagawa, Tsutomu Suzuki, Fumiaki Ebihara, Akio Protein Pept Lett Article BACKGROUND: Auto-induction is a convenient way to produce recombinant proteins without inducer addition using lac operon-controlled Escherichia coli expression systems. Auto-induction can occur unintentionally using a complex culture medium prepared by mixing culture substrates. The differences in culture substrates sometimes lead to variations in the induction level. OBJECTIVES: In this study, we investigated the feasibility of using glucose and lactose as boosters of auto-induction with a complex culture medium. METHODS: First, auto-induction levels were assessed by quantifying recombinant GFPuv expression under the control of the T7 lac promoter. Effectiveness of the additive-containing medium was examined using ovine angiotensinogen (tac promoter-based expression) and Thermus thermophilus manganese-catalase (T7 lac promoter-based expression). RESULTS: Auto-induced GFPuv expression was observed with the enzymatic protein digest Polypepton, but not with another digest tryptone. Regardless of the type of protein digest, supplementing Terrific Broth medium with glucose (at a final concentration of 2.9 g/L) and lactose (at a final concentration of 7.6 g/L) was successful in obtaining an induction level similar to that achieved with a commercially available auto-induction medium. The two recombinant proteins were produced in milligram quantity of purified protein per liter of culture. CONCLUSION: The medium composition shown in this study would be practically useful for attaining reliable auto-induction for E. coli-based recombinant protein production. Bentham Science Publishers 2020-08-05 2020-08-05 /pmc/articles/PMC8811614/ /pubmed/34353248 http://dx.doi.org/10.2174/0929866528666210805120715 Text en © 2021 Bentham Science Publishers https://creativecommons.org/licenses/by-nc/4.0/ This is an open access article licensed under the terms of the Creative Commons Attribution-Non-Commercial 4.0 International Public License (CC BY-NC 4.0) (https://creativecommons.org/licenses/by-nc/4.0/legalcode), which permits unrestricted, non-commercial use, distribution and reproduction in any medium, provided the work is properly cited. |
spellingShingle | Article Tahara, Nariyasu Tachibana, Itaru Takeo, Kazuyo Yamashita, Shinji Shimada, Atsuhiro Hashimoto, Misuzu Ohno, Satoshi Yokogawa, Takashi Nakagawa, Tsutomu Suzuki, Fumiaki Ebihara, Akio Boosting Auto-Induction of Recombinant Proteins in Escherichia coli with Glucose and Lactose Additives |
title | Boosting Auto-Induction of Recombinant Proteins in Escherichia coli with Glucose and Lactose Additives |
title_full | Boosting Auto-Induction of Recombinant Proteins in Escherichia coli with Glucose and Lactose Additives |
title_fullStr | Boosting Auto-Induction of Recombinant Proteins in Escherichia coli with Glucose and Lactose Additives |
title_full_unstemmed | Boosting Auto-Induction of Recombinant Proteins in Escherichia coli with Glucose and Lactose Additives |
title_short | Boosting Auto-Induction of Recombinant Proteins in Escherichia coli with Glucose and Lactose Additives |
title_sort | boosting auto-induction of recombinant proteins in escherichia coli with glucose and lactose additives |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8811614/ https://www.ncbi.nlm.nih.gov/pubmed/34353248 http://dx.doi.org/10.2174/0929866528666210805120715 |
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