Difference between revisions of "Part:BBa K2559000"

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We introduced bcsZ, H, A, B, C, D, bglX from the Acetobacter xylinum which involve in the process of bacterial cellulose synthesis into the model organism cyanobacteria (Synechocystis sp.pcc6803),and we obtain the transgenic cyanobacteria.
 
We introduced bcsZ, H, A, B, C, D, bglX from the Acetobacter xylinum which involve in the process of bacterial cellulose synthesis into the model organism cyanobacteria (Synechocystis sp.pcc6803),and we obtain the transgenic cyanobacteria.
  
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Figure 1
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Figure1
  
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Figure 2
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Figure2
 
Degradation of CMC by BcsZ
 
Degradation of CMC by BcsZ
 
The BcsZ in E. coli was even crystallized in a complex with the substrate cellopentaose.When BcsZ-overexpressing E. coli cells were grown on agar plates containing 2% CMC, a clear halo appeared around the colonies after staining with Congo Red.Purified cellulase from Aspergillus niger (Tokyo Chemical Industry, GenBank entry CAA03658.1) and bovine serum albumin served as positive and negative controls, respectively.[2]
 
The BcsZ in E. coli was even crystallized in a complex with the substrate cellopentaose.When BcsZ-overexpressing E. coli cells were grown on agar plates containing 2% CMC, a clear halo appeared around the colonies after staining with Congo Red.Purified cellulase from Aspergillus niger (Tokyo Chemical Industry, GenBank entry CAA03658.1) and bovine serum albumin served as positive and negative controls, respectively.[2]
  
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Figure 3
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Figure3
 
Cyanobacteria glucose content measurement (3 repeat). The same of quality of transferred cyanobacteria with bcsZH-ABCD-bglX gene and wildtype are treated with lysozyme to break out the cell. The glucose represents the content of cellulose. The addition of cellulase can digest the cellulose to glucose then the bacterial cellulose can measured. The distinct differences between the red column and blue column indicate the high expression intensity of bacteria cellulose. Wilcoxon test indicates our data is reliable.
 
Cyanobacteria glucose content measurement (3 repeat). The same of quality of transferred cyanobacteria with bcsZH-ABCD-bglX gene and wildtype are treated with lysozyme to break out the cell. The glucose represents the content of cellulose. The addition of cellulase can digest the cellulose to glucose then the bacterial cellulose can measured. The distinct differences between the red column and blue column indicate the high expression intensity of bacteria cellulose. Wilcoxon test indicates our data is reliable.
  
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Figure 4
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Figure4
  
  

Revision as of 09:29, 13 October 2018


Bacterial cellulose synthase Z (BcsZ) The BBa_K2559000 is an endoglucanase (Bcs Z) involved in the process of cellulose hydrolysis, and its roles in cellulose biosynthesis have long remained obscure.

Usage and Biology

Bacterial cellulose synthase Z (bcsZ) from several bacteria have been shown to possess an endo-β-1,4-glucanase (cellulase) activity, and E. coli protein was even crystallized in a complex with the substrate cellopentaose, however, the roles in cellulose biosynthesis have long remained obscure. We suspect that the BcsZ and BlgX may help the cell form the correct cellulose configuration through selective digestion.[1] We introduced bcsZ, H, A, B, C, D, bglX from the Acetobacter xylinum which involve in the process of bacterial cellulose synthesis into the model organism cyanobacteria (Synechocystis sp.pcc6803),and we obtain the transgenic cyanobacteria.

Figure1

Figure2 Degradation of CMC by BcsZ The BcsZ in E. coli was even crystallized in a complex with the substrate cellopentaose.When BcsZ-overexpressing E. coli cells were grown on agar plates containing 2% CMC, a clear halo appeared around the colonies after staining with Congo Red.Purified cellulase from Aspergillus niger (Tokyo Chemical Industry, GenBank entry CAA03658.1) and bovine serum albumin served as positive and negative controls, respectively.[2]

Figure3 Cyanobacteria glucose content measurement (3 repeat). The same of quality of transferred cyanobacteria with bcsZH-ABCD-bglX gene and wildtype are treated with lysozyme to break out the cell. The glucose represents the content of cellulose. The addition of cellulase can digest the cellulose to glucose then the bacterial cellulose can measured. The distinct differences between the red column and blue column indicate the high expression intensity of bacteria cellulose. Wilcoxon test indicates our data is reliable.

Figure4


The part is facilitate the in-depth research for other teams! Reference : 1.Romling U & Galperin MY (Bacterial cellulose biosynthesis: diversity of operons, subunits, products, and functions. (Translated from eng) Trends Microbiol 23(9):545-557 (in eng). 2.Mazur O & Zimmer J (Apo- and cellopentaose-bound structures of the bacterial cellulose synthase subunit BcsZ. (Translated from eng) J Biol Chem 286(20):17601-17606 (in eng).

The proposed organization of BcsZ in the BCS complexes

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
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 252