Part:BBa_K2559010

Bcs ABCD

The BBa K2559010 consist of bcsA、bcsB、bcsC、bcsD gene，which are coding the core subunits of bacterial cellulose synthase.

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

Figure 1 Proposed organization of the BCS complexes from Komagataeibacter xylinus

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 2 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