Team:Shanghai Metro HS/Contribution

Shanghai_Metro_HS

BBa_K4096001

== Profile ==

Name: PKC-001
Base Pairs: 1077bp
Origin: Pseudomonas aeruginosa, genome
Properties: A coding sequence of alkali cellulase.

==== Usage and Biology ====

BBa_K4096001 is a coding sequence of alkali cellulase (PKC-001) from Pseudomonas aeruginosa. Cellulases break down the cellulose molecule into monosaccharides ("simple sugars") such as beta-glucose, or shorter polysaccharides and oligosaccharides. Several different kinds of cellulases are known, which differ structurally and mechanistically. Synonyms, derivatives, and specific enzymes associated with the name "cellulase" include endo-1,4-beta-D-glucanase (beta-1,4-glucanase, beta-1,4-endoglucan hydrolase, endoglucanase D, 1,4-(1,3,1,4)-beta-D-glucan 4-glucanohydrolase), carboxymethyl cellulase (CMCase), avicelase, celludextrinase, cellulase A, cellulosin AP, alkali cellulase, cellulase A 3, 9.5 cellulase, and pancellase SS.

BBa_K4096002

== Profile ==

Name: PKC-OP
Base Pairs: 1086bp
Origin: Pseudomonas aeruginosa, genome
Properties: A coding sequence of alkali cellulase.

==== Usage and Biology ====

BBa_K4096002 is a coding sequence of alkali cellulase (PKC-OP) from Pseudomonas aeruginosa. Cellulases break down the cellulose molecule into monosaccharides ("simple sugars") such as beta-glucose, or shorter polysaccharides and oligosaccharides. Several different kinds of cellulases are known, which differ structurally and mechanistically. Synonyms, derivatives, and specific enzymes associated with the name "cellulase" include endo-1,4-beta-D-glucanase (beta-1,4-glucanase, beta-1,4-endoglucan hydrolase, endoglucanase D, 1,4-(1,3,1,4)-beta-D-glucan 4-glucanohydrolase), carboxymethyl cellulase (CMCase), avicelase, celludextrinase, cellulase A, cellulosin AP, alkali cellulase, cellulase A 3, 9.5 cellulase, and pancellase SS.

== Construct design ==

The alkaline cellulase gene from Pseudomonas aeruginosa PKC-001 was selected and we performed codon optimization on PKC-001 sequence to get PKC-OP, which was inserted into the pET-25b vector which contains a pelB signal peptide (Figure 1). The recombinant plasmid then was transformed it to E. coli BL21(DE3) strain to produce cellulase.

Figure 1. Genetic design of our gene

BBa_K4096004

== Profile ==

Name: pET-25b-PKC-OP
Base Pairs: 6633 bp
Origin: Synthetic
Properties: A recombinant plasmid containing cellulase sequence.

== Usage and Biology ==

BBa_K4096004 is a plasmid that can expressed cellulase (PKC-OP) under the control of T7 promoter from Pseudomonas aeruginosa. Cellulases break down the cellulose molecule into monosaccharides ("simple sugars") such as beta-glucose, or shorter polysaccharides and oligosaccharides. Several different kinds of cellulases are known, which differ structurally and mechanistically. Synonyms, derivatives, and specific enzymes associated with the name "cellulase" include endo-1,4-beta-D-glucanase (beta-1,4-glucanase, beta-1,4-endoglucan hydrolase, endoglucanase D, 1,4-(1,3,1,4)-beta-D-glucan 4-glucanohydrolase), carboxymethyl cellulase (CMCase), avicelase, celludextrinase, cellulase A, cellulosin AP, alkali cellulase, cellulase A 3, 9.5 cellulase, and pancellase SS.

== Construct design ==

The alkaline cellulase gene from Pseudomonas aeruginosa PKC-OP performed codon optimization on PKC-001 was selected and inserted into the pET-25b vector which contains a pelB signal peptide (Figure 1 and 2). The recombinant plasmid then was transformed it to E. coli BL21(DE3) strain to produce cellulase.

Figure 1. Genetic design of our gene

Figure 2. Schematic map of pET-25b-PKC expression plasmid.

BBa_K4096006

== Profile ==

Name: xynA-PKC-OP
Base Pairs: 1728bp
Origin: Sythetic
Properties: A coding sequence of xynA and PKC-OP.

== Usage and BioBBa_K4096006 is a coding sequence of alkali cellulase (PKC-OP) from Pseudomonas aeruginosa and xylanase (xynA) from Bacillus subtilis. Cellulases break down the cellulose molecule into monosaccharides ("simple sugars") such as beta-glucose, or shorter polysaccharides and oligosaccharides. Several different kinds of cellulases are known, which differ structurally and mechanistically. Xylanase and β-xylosidase are the backbone degrading enzymes of heterogeneous xylan, which can degrade the xylan backbone to produce xylo-oligosaccharides or xyloses with different degrees of polymerization, which play an important role in the degradation of hemicellulose. logy ==

== Construct design ==

Xylanase has the properties of hydrolyzing hemicellulose and can cooperate with cellulase to promote the biotransformation of lignocellulose. Add the Bacillus subtilis xylanase xynA gene to the cloning recombinant vector to make it compatible with cellulase PKC-OP and express β-xylosidase at the same time (as shown in the figure below).

Figure 1 Demonstration of Partnership (Mixed Feed)

The new family β-xylosidase from Humicola insolens Y1 has high tolerance to D-xylose. The gene is linked to a vector expressing xylanase xynA through GS linker to form pSIP403-PUS-xynA -xyl3A recombinant plasmid.

Figure 2. pSIP403-PUS-xynA -xyl3A