Recombinant protein production in Pseudoalteromonas haloplanktis TAC125 biofilm

IF 5.9 Q1 MICROBIOLOGY Biofilm Pub Date : 2024-01-24 DOI:10.1016/j.bioflm.2024.100179
Marzia Calvanese , Caterina D'Angelo , Concetta Lauro , Maria Luisa Tutino , Ermenegilda Parrilli
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Abstract

Biofilms have great potential for producing valuable products, and recent research has been performed on biofilms for the production of compounds with biotechnological and industrial relevance. However, the production of recombinant proteins using this system is still limited. The recombinant protein production in microbial hosts is a well-established technology and a variety of expression systems are available. Nevertheless, the production of some recombinant proteins can result in proteolyzed, insoluble, and non-functional forms, therefore it is necessary to start the exploration of non-conventional production systems that, in the future, could be helpful to produce some “difficult” proteins. Non-conventional production systems can be based on the use of alternative hosts and/or on non-conventional ways to grow recombinant cells. In this paper, the use of the Antarctic marine bacterium Pseudoalteromonas haloplanktis TAC125 grown in biofilm conditions was explored to produce two fluorescent proteins, GFP and mScarlet. The best conditions for the production were identified by working on media composition, and induction conditions, and by building a new expression vector suitable for the biofilm conditions. Results reported demonstrated that the optimized system for the recombinant protein production in biofilm, although it takes longer than planktonic production, has the same potentiality as the classical planktonic approach with additional advantages since it needs a lower concentration of the carbon sources and doesn't require antibiotic addition. Moreover, in the case of mScarlet, the production in biofilm outperforms the planktonic system in terms of a better quality of the recombinant product.

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卤化假单胞菌 TAC125 生物膜中的重组蛋白生产
生物膜在生产有价值的产品方面具有巨大潜力,最近已经开展了生物膜研究,以生产与生物技术和工业相关的化合物。然而,利用这一系统生产重组蛋白的研究仍然有限。在微生物宿主中生产重组蛋白是一项成熟的技术,有多种表达系统可供选择。然而,一些重组蛋白的生产可能会产生蛋白水解、不溶解和无功能的形式,因此有必要开始探索非常规生产系统,以便在未来帮助生产一些 "困难 "蛋白。非常规生产系统可以基于替代宿主的使用和/或重组细胞的非常规生长方式。本文探讨了利用在生物膜条件下生长的南极海洋假单胞菌 Haloplanktis TAC125 来生产两种荧光蛋白--GFP 和 mScarlet。通过研究培养基成分、诱导条件以及构建适合生物膜条件的新表达载体,确定了最佳生产条件。报告结果表明,生物膜重组蛋白生产的优化系统虽然比浮游生物生产耗时更长,但与传统的浮游生物方法具有相同的潜力,而且还具有额外的优势,因为它需要的碳源浓度更低,而且不需要添加抗生素。此外,就 mScarlet 而言,生物膜生产在提高重组产品的质量方面优于浮游生物系统。
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来源期刊
Biofilm
Biofilm MICROBIOLOGY-
CiteScore
7.50
自引率
1.50%
发文量
30
审稿时长
57 days
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