Difference between revisions of "Part:BBa K2559000"

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<partinfo>BBa_K2559000 short</partinfo>  
 
<partinfo>BBa_K2559000 short</partinfo>  
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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.
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The BBa_K2559000 encodes an endoglucanase (Bcs Z) that is responsible for the hydrolysis of cellulose. Its roles in cellulose biosynthesis still remained obscure.
  
 
===Usage and Biology===
 
===Usage and Biology===
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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]
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Bacterial cellulose synthase Z (BcsZ) in different bacteria species had been shown to function similarly to an endo-β-1,4-glucanase (cellulase). Nevertheless, BcsZ found in E. coli was even crystallized in a complex with the substrate cellopentaose. Its roles in cellulose biosynthesis still remain obscure. We suspect that the BcsZ and BlgX may responsible for the formation of the correct cellulose configuration through selective digestion.[1]
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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.
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[[File:Figure_1.png|400px|thumb|left| The proposed organization of the BCS complexes from Acetobacter xylinum. he pink color indicates the location of BcsZ in the plasmid membrane. [1]]]
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[[File:Scau-china-2018-2.png|400px|thumb|right|BcsZ adopts an (α/α)6-barrel fold. BcsZ is shown as a cartoon from a top (A) and side (B) view. The inner and outer helices are shown in dark and pale green, respectively, and the three β-sheets flanking the substrate binding cleft are shown in blue. The catalytic residues Glu55 and Asp243 are shown as red sticks. Helices are labeled H1–H12 from the N to the C terminus. The C-terminal residues 349–357 of BcsZ form a two-stranded β-sheet (colored orange) on the opposite side of the substrate binding pocket.[2]]]
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[[File:Scau-china-2018-2.png|800px|thumb|center|Figure 1 BcsZ adopts an (α/α)6-barrel fold. [2]]]
  
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Degradation of CMC by BcsZ
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===Degradation of CMC by BcsZ===
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[[File:Scau-china-2018-3.png|600px|thumb|center| The right show degradation of CMC by BcsZ which indicate the cellulase activity of BcsZ in E.coli.]]
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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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[[File:Scau-china-2018-3.png|800px|thumb|center|Figure 2 The right area show degradation of CMC by BcsZ which indicated the cellulase activity of BcsZ in E.coli.[2]]]
  
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Transgenic cyanobacteria with bcsZH-ABCD-bglX genes and the cellulose measurement
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We introduced ''bcs''Z, H, A, B, C, D, ''bgl''X from the ''Acetobacter xylinum ''which are involved in the process of bacterial cellulose synthesis into the cyanobacteria.
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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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[[File:Scau-china-2018-4.png|600px|thumb|center| The measurement of cellulose content in transgenic cyanobacteria which expressed bcs gene . The P-value verified that the distinction between treatment group  and control group. ]]
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===Transgenic cyanobacteria with bcsZH-ABCD-bglX genes and the cellulose measurement===
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Measurement of cyanobacteria glucose (3 repeats). The same of amount of transgenic cyanobacteria with bcsZH-ABCD-bglX genes and wild-type were treated with lysozyme to break the cells. Due to lacking a direct way to measure the content of cellu;ose in bacterial cell wall. Therefore, glucose can be used as an alternative parameter for measuring the content of cellulose since it can be digested into glucose by cellulose.
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The differences between the red colum and blue column indicated that the content of bacteria cellulose.
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[[File:Scau-china-2018-4.png|800px|thumb|center|Figure 3 The measurement of cellulose content in transgenic cyanobacteria which expressed bcs gene . The P-value verified that the distinction between treatment group  and control group. ]]
  
  
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Reference :
 
Reference :
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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).
 
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).
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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).
 
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).
  
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===The proposed organization of BcsZ in the BCS complexes ===
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Latest revision as of 09:04, 16 October 2018


Bacterial cellulose synthase Z (BcsZ)

The BBa_K2559000 encodes an endoglucanase (Bcs Z) that is responsible for the hydrolysis of cellulose. Its roles in cellulose biosynthesis still remained obscure.

Usage and Biology

Bacterial cellulose synthase Z (BcsZ) in different bacteria species had been shown to function similarly to an endo-β-1,4-glucanase (cellulase). Nevertheless, BcsZ found in E. coli was even crystallized in a complex with the substrate cellopentaose. Its roles in cellulose biosynthesis still remain obscure. We suspect that the BcsZ and BlgX may responsible for the formation of the correct cellulose configuration through selective digestion.[1]

Figure 1 BcsZ adopts an (α/α)6-barrel fold. [2]

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]

Figure 2 The right area show degradation of CMC by BcsZ which indicated the cellulase activity of BcsZ in E.coli.[2]

We introduced bcsZ, H, A, B, C, D, bglX from the Acetobacter xylinum which are involved in the process of bacterial cellulose synthesis into the cyanobacteria.

Transgenic cyanobacteria with bcsZH-ABCD-bglX genes and the cellulose measurement

Measurement of cyanobacteria glucose (3 repeats). The same of amount of transgenic cyanobacteria with bcsZH-ABCD-bglX genes and wild-type were treated with lysozyme to break the cells. Due to lacking a direct way to measure the content of cellu;ose in bacterial cell wall. Therefore, glucose can be used as an alternative parameter for measuring the content of cellulose since it can be digested into glucose by cellulose. The differences between the red colum and blue column indicated that the content of bacteria cellulose.

Figure 3 The measurement of cellulose content in transgenic cyanobacteria which expressed bcs gene . The P-value verified that the distinction between treatment group  and control group.


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).


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