Difference between revisions of "Part:BBa K2559002"

Latest revision as of 09:10, 16 October 2018

Bacterial cellulose synthase X(BglX)

The BBa_K2559001 is coding a β-glucosidase (BglX) which is involved in the bcs operons.

Usage and Biology

BglX is a β-glucosidase which participates the cyanoamino acid metabolism (ko00460), starch and sucrose metabolism (ko00500), phenylpropanoid biosynthesis (ko00940), metabolic pathways (ko01100), biosynthesis of secondary metabolites (ko01110). The enzyme is expected to participate in hydrolysis of bacterial cellulose, instead of synthesis. Its role in cellulose biosynthesis remains to be further explored.

Figure 1 The pathways involve bcsH（entry 3.2.1.21） in starch and sucrose metabolism(ko00500)

We transfered 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 c and blue column indicated that the content of bacteria cellulose.

Figure 2 The measurement of cellulose content in transgenic cyanobacteria with bcs gene.

The parts can further 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
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 516
Illegal XhoI site found at 1206
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 2099
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 339

@@ Line 7: / Line 7: @@
 ===Usage and Biology===
-BGLX is a β-glucosidase which participates the cyanoamino acid metabolism (ko00460), starch and sucrose metabolism (ko00500), phenylpropanoid biosynthesis (ko00940), metabolic pathways (ko01100), biosynthesis of secondary metabolites (ko01110).
+BglX is a β-glucosidase which participates the cyanoamino acid metabolism (ko00460), starch and sucrose metabolism (ko00500), phenylpropanoid biosynthesis (ko00940), metabolic pathways (ko01100), biosynthesis of secondary metabolites (ko01110).
 The enzyme is expected to participate in hydrolysis of bacterial cellulose, instead of synthesis. Its role in cellulose biosynthesis remains to be further explored.
-[[File:Scau-china-2018-10.png|800px|thumb|center|The pathways involve bcsH（entry 3.2.1.21） in Starch and sucrose metabolism(ko00500)]]
+[[File:Scau-china-2018-10.png|800px|thumb|center|Figure 1 The pathways involve bcsH（entry 3.2.1.21） in starch and sucrose metabolism(ko00500)]]
-We transfered 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.
-Transgenic cyanobacteria with bcsZH-ABCD-bglX genes and the cellulose measurement
+We transfered ''bcsZ'',H, A, B, C, D, bgl''X'' from the ''Acetobacter xylinum'' which are involved in the process of bacterial cellulose synthesis into the cyanobacteria.
-Cyanobacteria glucose content measurement (3 repeats). 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.
-[[File:Scau-china-2018-4.png|800px|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. ]]
+===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 c and blue column indicated that the content of bacteria cellulose.
+[[File:Scau-china-2018-4.png|800px|thumb|center|Figure 2 The measurement of cellulose content in transgenic cyanobacteria with bcs gene. ]]
+The parts can further facilitate the in-depth research for other teams!
+Reference :
+.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).
+.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).
 <!-- -->