Construction of a High-Expression System in Bacillus through Transcriptomic Profiling and Promoter Engineering

Miao, Cuicui; Liang, Han; Lu, Yao; Hong, Feng

doi:10.3390/microorganisms8071030

Cited by 14 publications

(9 citation statements)

References 56 publications

(75 reference statements)

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“…However, screening and generating promoter libraries is a labor-and time-consuming task. Thus, several recent studies have used more efficient RNA-seq to determine strong promoters [26,43]. Han L, et al have investigated the transcriptome of B. pumilus BA06 [27].…”

Section: Discussionmentioning

confidence: 99%

Novel sinIR promoter for Bacillus subtilis DB104 recombinant protein expression system

Jun

Kim

Hong

2023

JABC

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Transcriptome analysis revealed that the sinR gene encoding a transition-state regulator of Bacillus pumilus, genetically close to B. subtilis, was expressed at high levels during growth. The sinR gene is the second gene of the sinIR operon consisting of three promoters and two structural genes in B. subtilis. This study used the sinIR promoter of B. subtilis DB104 to construct a recombinant protein expression system. First, the expression ability depending on the number of sinIR promoter was investigated using enhanced green fluorescent protein (eGFP). The expression level of eGFP was slightly higher when using two promoters (Psin2) than using original promoters. The Psin2 promoter was further engineered by modifying the repressor binding site and 35 and 10 regions. Shine-Dalgarno (SD) sequence of the sinI gene was modified to the consensus sequence. Finally, combining the engineered Psin2 promoter with the modified SD sequence increased the expression level of eGFP by about 13.4-fold over the original promoter. Our results suggest that the optimized sinIR promoter could be used as a novel tool for recombinant protein expression in B. subtilis.

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Section: Discussionmentioning

confidence: 99%

Novel sinIR promoter for Bacillus subtilis DB104 recombinant protein expression system

Jun

Kim

Hong

2023

JABC

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“…One of the most cost-effective and efficient methods to achieve high-level production of recombinant proteins remains the optimization of the promoter at the level of transcription. Transcriptomic data are often used for the identification of strong promoters, either native or heterologous, and promoter engineering, including semi-rational evolution, is a powerful tool for increasing the target protein production (Miao et al 2020 ). Some of the promoters used for recombinant production of industrially relevant enzymes in Bacillus and Streptomyces spp.…”

Section: Molecular Tools Developed For Increased Production Of Recomb...mentioning

confidence: 99%

Bacillus and Streptomyces spp. as hosts for production of industrially relevant enzymes

Vojnovic,

Aleksic,

Ilic-Tomic

et al. 2024

Appl Microbiol Biotechnol

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The application of enzymes is expanding across diverse industries due to their nontoxic and biodegradable characteristics. Another advantage is their cost-effectiveness, reflected in reduced processing time, water, and energy consumption. Although Gram-positive bacteria, Bacillus, and Streptomyces spp. are successfully used for production of industrially relevant enzymes, they still lag far behind Escherichia coli as hosts for recombinant protein production. Generally, proteins secreted by Bacillus and Streptomyces hosts are released into the culture medium; their native conformation is preserved and easier recovery process enabled. Given the resilience of both hosts in harsh environmental conditions and their spore-forming capability, a deeper understanding and broader use of Bacillus and Streptomyces as expression hosts could significantly enhance the robustness of industrial bioprocesses. This mini-review aims to compare two expression hosts, emphasizing their specific advantages in industrial surroundings such are chemical, detergent, textile, food, animal feed, leather, and paper industries. The homologous sources, heterologous hosts, and molecular tools used for the production of recombinant proteins in these hosts are discussed. The potential to use both hosts as biocatalysts is also evaluated. Undoubtedly, Bacillus and Streptomyces spp. as production hosts possess the potential to take on a more substantial role, providing superior (bio-based) process robustness and flexibility. Key points • Bacillus and Streptomyces spp. as robust hosts for enzyme production. • Industrially relevant enzyme groups for production in alternative hosts highlighted. • Molecular biology techniques are enabling easier utilization of both hosts.

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“…Based on this concept, the group patented and built Somartex R (self operating marine trapping extractor) for testing in different marine locations 13 . Alternatively, compounds of interest can be produced with plasmid-based expression systems in well-established prokaryotic and eukaryotic hosts, such as Bacillus subtilis, Escherichia coli, Saccharomyces cerevisiae (Kroll et al, 2010;Mao et al, 2017;Miao et al, 2020), the yeast genus Pichia as well as cell systems. These recombinant expression systems have dramatically increased the opportunities for biotechnology and commercialization of products.…”

Section: Production Upscalingmentioning

confidence: 99%

The Essentials of Marine Biotechnology

Barbier²,

et al. 2021

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Coastal countries have traditionally relied on the existing marine resources (e.g., fishing, food, transport, recreation, and tourism) as well as tried to support new economic endeavors (ocean energy, desalination for water supply, and seabed mining). Modern societies and lifestyle resulted in an increased demand for dietary diversity, better health and well-being, new biomedicines, natural cosmeceuticals, environmental conservation, and sustainable energy sources. These societal needs stimulated the interest of researchers on the diverse and underexplored marine environments as promising and sustainable sources of biomolecules and biomass, and they are addressed by the emerging field of marine (blue) biotechnology. Blue biotechnology provides opportunities for a wide range of initiatives of commercial interest for the pharmaceutical, biomedical, cosmetic, nutraceutical, food, feed, agricultural, and related industries. This article synthesizes the essence, opportunities, responsibilities, and challenges encountered in marine biotechnology and outlines the attainment and valorization of directly derived or bio-inspired products from marine organisms. First, the concept of bioeconomy is introduced. Then, the diversity of marine bioresources including an overview of the most prominent marine organisms and their potential for biotechnological uses are described. This is followed by introducing methodologies for exploration of these resources and the main use case scenarios in energy, food and feed, agronomy, bioremediation and climate change, cosmeceuticals, bio-inspired materials, healthcare, and well-being sectors. The key aspects in the fields of legislation and funding are provided, with the emphasis on the importance of communication and stakeholder engagement at all levels of biotechnology development. Finally, vital overarching concepts, such as the quadruple helix and Responsible Research and Innovation principle are highlighted as important to follow within the marine biotechnology field. The authors of this review are collaborating under the European Commission-funded Cooperation in Science and Technology (COST) Action Ocean4Biotech – European transdisciplinary networking platform for marine biotechnology and focus the study on the European state of affairs.

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Construction of a High-Expression System in Bacillus through Transcriptomic Profiling and Promoter Engineering

Cited by 14 publications

References 56 publications

Novel sinIR promoter for Bacillus subtilis DB104 recombinant protein expression system

Novel sinIR promoter for Bacillus subtilis DB104 recombinant protein expression system

Bacillus and Streptomyces spp. as hosts for production of industrially relevant enzymes

The Essentials of Marine Biotechnology

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