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Related Concept Videos

Gene Regulation During Sporulation01:17

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Sporulation is a complex developmental process that allows certain Gram-positive bacteria, such as Bacillus subtilis and Clostridium species, to survive extreme environmental conditions. This process is tightly regulated by a series of signaling cascades and transcriptional controls, ensuring the formation of a highly resistant endospore.Sporulation is triggered by unfavorable conditions, such as nutrient depletion, and is governed by a phosphorelay system. One of the sensor kinases, such as...
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Cohesin protein complexes are a molecular glue that holds two sister chromatids together. They play an important role both in mitosis and meiosis. In mitosis, all cohesin complexes present on the chromosomes are removed before the start of the anaphase stage.
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Groups of proteins may form a complex where each protein in this complex has a different role in the overall execution of the complex’s function. Often some of the proteins in the complex can be replaced by a closely related variant to give a complex that contains many of the same components yet is functionally distinct.
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Bacterial cells were initially considered simple, randomly organized structures lacking a cytoskeleton. However, the discovery of cytoskeleton homologs in bacteria led to the change of this opinion. Bacterial cytoskeletal filaments regulate the cell shape, cell polarity, cell division, and partitioning of plasmids during cell division. It was later discovered that bacterial cytoskeletal proteins, mainly actin and tubulin homologs, are diverse compared to their eukaryotic counterparts. On the...
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Spore Adsorption as a Nonrecombinant Display System for Enzymes and Antigens
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Constructing an Efficient Bacillus subtilis Spore Display by Using Cohesin-Dockerin Interactions.

He Wang1, Xiaomin Jiang2, Yongchang Qian3

  • 1School of Grain Science and Technology, Jiangsu University of Science and Technology, Zhenjiang 212100, China.

Molecules (Basel, Switzerland)
|March 6, 2021
PubMed
Summary
This summary is machine-generated.

Researchers enhanced Bacillus subtilis spore display efficiency using cohesin-dockerin modules. This cellulosome-inspired method improves enzyme immobilization on bacterial spores for potential applications.

Keywords:
Bacillus subtiliscohesin dockerin interactiondisplay efficiencyspore displayβ-galactosidase

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Area of Science:

  • Biotechnology
  • Microbial Engineering
  • Protein Display Systems

Background:

  • Bacillus subtilis spore display is a growing area of interest for biotechnological applications.
  • Improving the efficiency of protein display on bacterial spores is crucial for their wider use.
  • Cellulosome architecture offers a model for targeted enzyme immobilization.

Purpose of the Study:

  • To enhance Bacillus subtilis spore display efficiency using a cellulosome-inspired cohesin-dockerin (Coh-Doc) system.
  • To evaluate the display capabilities of five different Coh-Doc pairs for immobilizing β-galactosidase.
  • To confirm surface expression and functionality of the displayed enzyme.

Main Methods:

  • Introduction of divergent cohesin modules onto Bacillus subtilis spores.
  • Specific binding of target enzymes engineered with matching dockerin modules.
  • Confirmation of surface expression via immunofluorescence microscopy, western blotting, and dot blotting.
  • Enzyme activity assay using o-nitrophenyl-β-D-galactopyranoside substrate.

Main Results:

  • Five different cohesin-dockerin pairs were successfully utilized for displaying β-galactosidase on Bacillus subtilis spores.
  • All engineered spores demonstrated hydrolytic activity, confirming successful enzyme display and functionality.
  • An optimized Coh-Doc based spore display system achieved the highest display efficiency.

Conclusions:

  • The cohesin-dockerin interaction provides an effective strategy for enhancing Bacillus subtilis spore display.
  • This cellulosome-inspired approach offers a versatile platform for immobilizing enzymes on bacterial spores.
  • The findings open new perspectives for improving the efficiency and application scope of Bacillus subtilis spore display technology.