Difference between revisions of "Part:BBa K2559010"
Line 1: Line 1:
  
 
__NOTOC__
 
__NOTOC__
−
<partinfo>BBa_K2559000 short</partinfo>  
+
<partinfo>BBa_K2559010 short</partinfo>  
 +
 
  
−
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===
−
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]
 
−
 
−
[[File:Scau-china-2018-2.png|800px|thumb|center|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]
 
−
[[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]]]
 
  
 +
BcsA and BcsB are two catalytic subunits that are present in all BCS enzymes experimentally characterized so far.BcsC is a periplasmic protein that consists of an N-terminal α-helical part formed by several tetratricopeptide repeat (TPR) domains and a C-terminal part that is structurally similar to the β-barrels of outer membrane proteins.The TPR-containing N-terminal part of BcsC is believed to interact with peptidoglycan and other BSC components, while its C-terminal β-barrel domain is likely located in the outer membrane, forming a channel that guides the nascent glucan out of the cell.
 +
BcsD is a periplasmic protein that oligomerizes to form a cylindrical octamer ∼90 Å in diameter with a huge central pore that can accommodate up to four separate glucan molecules. BcsD seems to be required for the arrangement of the BCS complex along the longitudinal cell axis
 
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.
 
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.
  
Line 25: Line 19:
  
  
−
The part is facilitate the in-depth research for other teams!
+
 
  
 
Reference :
 
Reference :
Line 36: Line 30:
 
<!-- -->
 
<!-- -->
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
−
<partinfo>BBa_K2559000 SequenceAndFeatures</partinfo>
+
<partinfo>BBa_K2559010 SequenceAndFeatures</partinfo>
  
  
 
<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  
 
===Functional Parameters===
 
===Functional Parameters===
−
<partinfo>BBa_K2559000 parameters</partinfo>
+
<partinfo>BBa_K2559010 parameters</partinfo>
 
<!-- -->
 
<!-- -->

Revision as of 11:16, 17 October 2018


Bcs ABCD


Usage and Biology

BcsA and BcsB are two catalytic subunits that are present in all BCS enzymes experimentally characterized so far.BcsC is a periplasmic protein that consists of an N-terminal α-helical part formed by several tetratricopeptide repeat (TPR) domains and a C-terminal part that is structurally similar to the β-barrels of outer membrane proteins.The TPR-containing N-terminal part of BcsC is believed to interact with peptidoglycan and other BSC components, while its C-terminal β-barrel domain is likely located in the outer membrane, forming a channel that guides the nascent glucan out of the cell. BcsD is a periplasmic protein that oligomerizes to form a cylindrical octamer ∼90 Å in diameter with a huge central pore that can accommodate up to four separate glucan molecules. BcsD seems to be required for the arrangement of the BCS complex along the longitudinal cell axis 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.



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
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 224
    Illegal BglII site found at 1136
    Illegal BamHI site found at 4237
    Illegal XhoI site found at 232
    Illegal XhoI site found at 3313
    Illegal XhoI site found at 4066
    Illegal XhoI site found at 6940
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 212
    Illegal NgoMIV site found at 5214
    Illegal NgoMIV site found at 7528
    Illegal NgoMIV site found at 8035
    Illegal AgeI site found at 2187
    Illegal AgeI site found at 4292
    Illegal AgeI site found at 7309
    Illegal AgeI site found at 7598
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 4063