Coding

Part:BBa_K4591003

Designed by: Jingtong Wang   Group: iGEM23_ZJUT-China   (2023-06-05)
Revision as of 10:57, 12 October 2023 by Kaga (Talk | contribs)

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Mistic (Mstx)

A membrane-integrating protein for membrane protein expression in Bacillus subtilis.

Usage and Biology

Mistic（MstX） is a unique protein discovered in a select few bacterial species, including Bacillus subtilis. When fused to the N-terminus of a cargo protein construct, it enables high-level heterologous expression and targets the complete membrane protein sequence to the cell membrane. Despite its small size and high acidity, Mistic associates with membranes, likely through autonomous binding to phospholipid bilayers, thereby bypassing or facilitating conventional secretion machinery.

The sequence and NMR structure of Mistic have been deposited in the NCBI protein database by TARMO P. ROOSILD et al. Matthew E. Lundberg, Eric C. Becker, Senyon Choe explored the mechanism by which MstX promotes the synthesis of the biofilm in Bacillus subtilis. Through mechanisms like regulating potassium ion channels and activating regulatory factors, MstX enhances biofilm formation in B. subtilis.

Fig 1. A positive autoregulatory loop involving MstX, YugO, potassium, and biofilm formation in B. subtilis.[1]

We have found that this protein is highly suitable for use in our attachment module, enabling engineered bacteria to effectively adhere to PET plastic, thus enhancing degradation efficiency.

Characterization

Fig 2. Alterations in colony morphology and biofilm formation related to mstX expression in the domesticated strain PY79 (A) Colony morphology of B. subtilis PY79. (B) Colony morphology of B. subtilis PY79 ΔmstX (MEL64). (C) Colony morphology of B. subtilis PY79 domesticated strain after IPTG induction of mstX (lacA::Pspac-mstX-erm; MEL66). (D) Colony morphology of Bacillus subtilis PY79 after IPTG induction of mstX (M75A) (lacA::Pspac-mstX (M75A)-erm; MEL67). (E) Microtitre crystal violet staining assay for WT, ΔmstX, lacA::Pspac –mstX-erm, and lacA::Pspac -mstX (M75A)-erm strains (strains PY79, MEL64, MEL66 and MEL67)[2].

The data indicates that MstX can effectively promote the formation of biofilms in Bacillus subtilis.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Parameters
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