Coding

Part:BBa_K2328068

Designed by: Zhongyi Jiang   Group: iGEM17_TJU_China   (2017-10-22)


AcmA (N-terminal)

The AcmA protein consists of two domains, an N-terminal catalytic domain including signal peptide (S.P.) and an active site (A.S.), and a C-terminal anchoring domain. The anchoring domain contains three repeats (LysM1-3) that are highly homologous.N-Acetylmuraminidase AcmA is an autolysin of L. lactis which is required for cell separation and is responsible for cell lysis during the stationary phase (Buist et al. 1995) The major peptidoglycan hydrolase (AcmA) of L. lactis consists of three domains: the N-terminal signal sequence, followed by an active domain and a C-terminal membrane anchor. The acmA gene encodes a polypeptide of 437 amino acids with a deduced molecular weight of 46 kDa (Buist 1997). Three repeated regions comprising 44 amino acids are present in the C-terminal domain of the protein. These repeats are involved in bacterial cell wall binding and are separated by intervening sequences which are highly enriched with serine, threonine and asparagine residues (Buist 1997). Only one of these repeats is sufficient for cell wall binding (Buist 1997). Here, we report on the expression and purification of one unit of the AcmA C-terminal repeat fragment from a recombinant Escherichia coli and describe a novel display method of this cell wall-binding fragment with viral epitopes exposed on the cell surface of L. lactis, Lactobacillus and Bacillus sphericus.

Usage and Biology

Acma and smURFP are joined together, and the fragment is inserted into pMG36e to construct a new plasmid. When the plasmid is expressed, smURFP is anchored to the cell surface by Acma, thus expressing the shape.

Reference

Li X, et al. Functional cell surface display of endo-beta-1, 3-1, 4-glucanase in Lactococcus lactis using N-acetylmuraminidase as the anchoring motif.Sheng Wu Gong Cheng Xue Bao. 2009.

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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Categories
Parameters
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