ArticleMicrobial cell factories2016
Homogeneity and heterogeneity in amylase production by Bacillus subtilis under different growth conditions.
Article in Microbial cell factories, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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Who cites it
21 citing papers in PubMed, 41 citations in OpenAlex.
- Exploration of Length and Hydrophobicity Requirements for Optimal α-Amylase Secretion in Bacillus subtilis Based on Rational Design.Microbial biotechnology · 2026Article
- Toward food-grade production of the Bacteroides helcogenes protein-glutamine glutaminase with an optimized Bacillus subtilis strain.Applied microbiology and biotechnology · 2026Article
- Impact of expression systems on the transcriptome ofApplied and environmental microbiology · 2025Article
- An inverse relationship between fitness and secretion efficiency in a gram-positive bacterium.PNAS nexus · 2025Article
- Efficient expression of γ-glutamyl transpeptidase in Bacillus subtilis via CRISPR/Cas9n and its immobilization.Applied microbiology and biotechnology · 2024Article
- TerC proteins function during protein secretion to metalate exoenzymes.Nature communications · 2023Article
- TerC Proteins Function During Protein Secretion to Metalate Exoenzymes.Research square · 2023Article
- TerC Proteins Function During Protein Secretion to Metalate Exoenzymes.bioRxiv : the preprint server for biology · 2023Article
- The ins and outs of Bacillus proteases: activities, functions and commercial significance.FEMS microbiology reviews · 2022Review
- An overview and future prospects of recombinant protein production in Bacillus subtilis.Applied microbiology and biotechnology · 2021Review
- Effect of Photocaged Isopropyl β-d-1-thiogalactopyranoside Solubility on the Light Responsiveness of LacI-controlled Expression Systems in Different Bacteria.Chembiochem : a European journal of chemical biology · 2021Article
- Relative contributions of non-essential Sec pathway components and cell envelope-associated proteases to high-level enzyme secretion by Bacillus subtilis.Microbial cell factories · 2020Article
- Biocatalytic Degradation Efficiency of Postconsumer Polyethylene Terephthalate Packaging Determined by Their Polymer Microstructures.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2019Article
- Proteases HtrA and HtrB for α-amylase secreted from Bacillus subtilis in secretion stress.Cell stress & chaperones · 2019Review
- Influence of global gene regulatory networks on single cell heterogeneity of green fluorescent protein production in Bacillus subtilis.Microbial cell factories · 2018Article
- Proteomic Analysis of Normal Expression Differences Exist in Bacillus Subtilis 168 Cultivation.Current microbiology · 2018Article
- Bottleneck in secretion of α-amylase in Bacillus subtilis.Microbial cell factories · 2017Review
- Technological Microbiology: Development and Applications.Frontiers in microbiology · 2017Review
- GlnR and PhoP Directly Regulate the Transcription of Genes Encoding Starch-Degrading, Amylolytic Enzymes in Saccharopolyspora erythraea.Applied and environmental microbiology · 2016Article
- The fed-batch principle for the molecular biology lab: controlled nutrient diets in ready-made media improve production of recombinant proteins in Escherichia coli.Microbial cell factories · 2016Review
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Authors and funding
9 authors at 4 institutions in 4 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundBacillus subtilis is an important cell factory for the biotechnological industry due to its ability to secrete commercially relevant proteins in large amounts directly into the growth medium. However, hyper-secretion of proteins, such as α-amylases, leads to induction of the secretion stress-responsive CssR-CssS regulatory system, resulting in up-regulation of the HtrA and HtrB proteases. These proteases degrade misfolded proteins secreted via the Sec pathway, resulting in a loss of product. The aim of this study was to investigate the secretion stress response in B. subtilis 168 cells overproducing the industrially relevant α-amylase AmyM from Geobacillus stearothermophilus, which was expressed from the strong promoter P(amyQ)-M.
resultsHere we show that activity of the htrB promoter as induced by overproduction of AmyM was "noisy", which is indicative for heterogeneous activation of the secretion stress pathway. Plasmids were constructed to allow real-time analysis of P(amyQ)-M promoter activity and AmyM production by, respectively, transcriptional and out-of-frame translationally coupled fusions with gfpmut3. Our results show the emergence of distinct sub-populations of high- and low-level AmyM-producing cells, reflecting heterogeneity in the activity of P(amyQ)-M. This most likely explains the heterogeneous secretion stress response. Importantly, more homogenous cell populations with regard to P(amyQ)-M activity were observed for the B. subtilis mutant strain 168degUhy32, and the wild-type strain 168 under optimized growth conditions.
conclusionExpression heterogeneity of secretory proteins in B. subtilis can be suppressed by degU mutation and optimized growth conditions. Further, the out-of-frame translational fusion of a gene for a secreted target protein and gfp represents a versatile tool for real-time monitoring of protein production and opens novel avenues for Bacillus production strain improvement.
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