A Review on Recent Advancement in Expression Strategies Used in
<i>Bacillus subtilis</i>

Ejaz, Sharoon; Khan, Hamza; Sarwar, Nadia; Aqeel, Sahibzada Muhammad; Al‐Adeeb, Abdulqader; Liu, Song

doi:10.2174/0929866529666220803163335

Cited by 5 publications

(5 citation statements)

References 80 publications

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“…Bacillus subtilis is a robust and efficient heterologous host that is generally used industrially for heterologous expression (Yang et al, 2021;Ejaz et al, 2022). A number of canonical NRPS pathways come from B. subtilis and other closely related Firmicutes including lipopeptides like surfactin (Sen, 2010), polymyxins (Komura and Kurahashi, 1985;Choi et al, 2009;Kim et al, 2015), bacillaene (Butcher et al, 2007), and plipastatin (Gao et al, 2018).…”

Section: Bacillus Sppmentioning

confidence: 99%

Cell-free protein synthesis for nonribosomal peptide synthetic biology

Sword,

Abbas,

Bailey

2024

Front. Nat. Prod.

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Peptide natural products have a wide range of useful applications as pesticides, veterinary agents, pharmaceuticals, and bioproducts. To discover new natural products, manipulate them for analog generation, and to harness the potential of these bioactive compounds for synthetic biology, it is necessary to develop robust methods for the expression of biosynthetic genes. Cell-free synthetic biology is emerging as an important complementary approach because it is highly desirable to express protein on a more rapid timescale and does not rely upon the genetic tractability of a strain thus improving the throughput of design-build-test-learn cycles. Additionally, generating metabolites outside the cell can overcome issues such as cellular toxicity which can hamper applications like antibiotic development. In this review, we focus on the cell-free production of peptide natural products generated by non-ribosomal peptide synthetase. Nonribsomal peptides are biosynthesized by non-ribosomal peptide synthetases which are large “mega” enzymes that provide specific challenges to heterologous expression. First, we summarize NRPSs and their corresponding peptide metabolites that are expressed in cell-free systems. With that, we discuss the requirements and challenges to express such large proteins in cell-free protein synthesis as well as host machineries that have been developed for cell-free protein synthesis that could be particularly relevant to generating non-ribosomal peptide metabolites in the future. The development of cell-free systems can then be used for prototyping to accelerate efforts towards engineered biosynthesis of these complex pathways.

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Section: Bacillus Sppmentioning

confidence: 99%

Cell-free protein synthesis for nonribosomal peptide synthetic biology

Sword,

Abbas,

Bailey

2024

Front. Nat. Prod.

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“…[12] B. subtilis, a well-known GRAS microorganism, is extensively used for industrial enzyme production due to its robust protein secretion, rapid growth rate, and suitability for scale-up fermentation. [13][14][15] However, the requirement of antibiotics for plasmid maintenance in B. subtilis raises safety concerns. This issue restricts the application of B. subtilis strains for SIase production in the food industry.…”

Section: Introductionmentioning

confidence: 99%

“…[19] In addition to the choice of expression host strain, the promoter in the expression vector also plays a pivotal role in modulating the expression of heterologous proteins. [14] The selection of a robust and controllable promoter is essential. Constitutive promoters such as P 43 , P HpaII , and P amyE have been used to direct recombinant protein expression in B. subtilis.…”

Section: Introductionmentioning

confidence: 99%

Development of an engineered Bacillus subtilis strain for antibiotic‐free sucrose isomerase production

Li,

Ren,

et al. 2024

Biotechnology Journal

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Sucrose isomerase (SIase) catalyzes the hydrolysis and isomerization of sucrose into isomaltulose, a functional sugar extensively used in the food industry. However, the lack of safe and efficient heterologous expression systems for SIase has constrained its production and application. In this study, an engineered Bacillus subtilis strain for antibiotic‐free SIase production was developed via a food‐grade expression system. First, the B. subtilis strain TEA was modified through the CRISPR/Cas9 system, resulting in a mutant strain TEA4, which exhibited enhanced capabilities for recombinant protein expression. For efficient and safe production of SIase, different constitutive and inducible promoters were evaluated. The maltose‐inducible promoter Poglv was found to have an extracellular SIase activity of 21.7 U mL‐1 in engineered strain TEA4. Subsequent optimization of the culture medium further increased SIase activity to 26.4 U mL‐1 during shake flask cultivation. Eventually, using the crude enzyme solution of the engineered strain in biotransformation reactions resulted in a high yield of isomaltulose under high concentrations sucrose, achieving a maximum yield of 83.1%. These findings demonstrated an engineered B. subtilis strain for antibiotic‐free SIase production, paving the way for its scale‐up industrial production and application.

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“…Bacillus subtilis has many attributes such as food security, easy genetic manipulation, and efficient protein secretion ability, which make it a popular host for recombinant protein production [1,2]. It also has long been used as a model organism for molecular research, as well as an industrial workhorse for the production of valuable enzymes [3].…”

Section: Introductionmentioning

confidence: 99%

N-Terminal Sequences of Signal Peptides Assuming Critical Roles in Expression of Heterologous Proteins in Bacillus subtilis

Zhang,

Zhen,

Teng

et al. 2024

Microorganisms

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The N-terminal sequences of proteins and their corresponding encoding sequences may play crucial roles in the heterologous expression. In this study, the secretory expression of alkaline pectin lyase APL in B. subtilis was investigated to explore the effects of the N-terminal 5–7 amino acid sequences of different signal peptides on the protein expression and secretion. It was identified for the first time that the first five amino acid sequences of the N-terminal of the signal peptide (SP-LipA) from Bacillus subtilis lipase A play an important role in promoting the expression of APL. Furthermore, it was revealed that SP-LipA resulted in higher secretory expression compared to other signal peptides in this study primarily due to its encoding of N-terminal amino acids with relatively higher transcription levels and its efficient secretion capacity. Based on this foundation, the recombinant strain constructed in this work achieved a new record for the highest extracellular yields of APL in B. subtilis, reaching 12,295 U/mL, which was 1.9-times higher than that expressed in the recombinant Escherichia coli strain previously reported. The novel theories uncovered in this study are expected to play significant roles in enhancing the expression of foreign proteins both inside and outside of cells.

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A Review on Recent Advancement in Expression Strategies Used in Bacillus subtilis

Cited by 5 publications

References 80 publications

Cell-free protein synthesis for nonribosomal peptide synthetic biology

Cell-free protein synthesis for nonribosomal peptide synthetic biology

Development of an engineered Bacillus subtilis strain for antibiotic‐free sucrose isomerase production

N-Terminal Sequences of Signal Peptides Assuming Critical Roles in Expression of Heterologous Proteins in Bacillus subtilis

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