Journal: Biotechnology for Biofuels

Article Title: Discovery of new cellulases from the metagenome by a metagenomics-guided strategy

doi: 10.1186/s13068-016-0557-3

Figure Lengend Snippet: PCR amplification of three cellulase genes. Lanes : 1, marker; 2, cel7482; 3, control (ddH 2 O as template); 4, marker; 5, cel3623; 6, control; 7, marker; 8, cel36; 9, control

Article Snippet: The nucleotide sequence encoding the twin-arginine signal peptide of B . subtilis YwbN [ ] and codon-optimized cel7482 was chemically synthesized by BGI Inc., Beijing, China.

Techniques: Amplification, Marker

Journal: Biotechnology for Biofuels

Article Title: Discovery of new cellulases from the metagenome by a metagenomics-guided strategy

doi: 10.1186/s13068-016-0557-3

Figure Lengend Snippet: SDS-PAGE analysis for the expression of the cellulase genes in E. coli BL21 (DE3). Lane 1 , total proteins from recombinant E. coli BL21 cells without IPTG induction; lane 2 , total proteins from recombinant E. coli BL21 cells induced by IPTG for 4 h; lane 3 , purified cellulase; M, protein molecular weight markers. a cel7482; b cel3623; c cel36

Article Snippet: The nucleotide sequence encoding the twin-arginine signal peptide of B . subtilis YwbN [ ] and codon-optimized cel7482 was chemically synthesized by BGI Inc., Beijing, China.

Techniques: SDS Page, Expressing, Recombinant, Purification, Molecular Weight

Journal: Biotechnology for Biofuels

Article Title: Discovery of new cellulases from the metagenome by a metagenomics-guided strategy

doi: 10.1186/s13068-016-0557-3

Figure Lengend Snippet: Purification of the recombinant His 6 -tagged cel7482, cel3623 and cel36 from 500 ml of E . coli cultures using immobilized metal ion affinity chromatography

Article Snippet: The nucleotide sequence encoding the twin-arginine signal peptide of B . subtilis YwbN [ ] and codon-optimized cel7482 was chemically synthesized by BGI Inc., Beijing, China.

Techniques: Purification, Recombinant, Activity Assay

Journal: Biotechnology for Biofuels

Article Title: Discovery of new cellulases from the metagenome by a metagenomics-guided strategy

doi: 10.1186/s13068-016-0557-3

Figure Lengend Snippet: HPAEC analysis for final products of hydrolysis of CMC by the recombinant cel7482, cel3623 and cel36. HPAEC analysis was performed on a Dionex ICS5000 system equipped with a pulsed amperometric detector and a CarboPac PA200 column (Dionex). a cel7482; b cel3623; c cel36

Article Snippet: The nucleotide sequence encoding the twin-arginine signal peptide of B . subtilis YwbN [ ] and codon-optimized cel7482 was chemically synthesized by BGI Inc., Beijing, China.

Techniques: Recombinant

Journal: Biotechnology for Biofuels

Article Title: Discovery of new cellulases from the metagenome by a metagenomics-guided strategy

doi: 10.1186/s13068-016-0557-3

Figure Lengend Snippet: a The effect of pH on activity of the recombinant cel7482, cel3623 and cel36. b The effect of temperature on activity of the recombinant cel7482, cel3623 and cel36

Article Snippet: The nucleotide sequence encoding the twin-arginine signal peptide of B . subtilis YwbN [ ] and codon-optimized cel7482 was chemically synthesized by BGI Inc., Beijing, China.

Techniques: Activity Assay, Recombinant

Journal: Biotechnology for Biofuels

Article Title: Discovery of new cellulases from the metagenome by a metagenomics-guided strategy

doi: 10.1186/s13068-016-0557-3

Figure Lengend Snippet: Thermostability of the recombinant cel7482, cel3623 and cel36. Each purified enzyme was incubated at different temperatures for different periods of time before determining the residual activity at optimal temperature in 50 mM citrate–phosphate buffer (pH 5.5) with CMC as substrate. a cel7482; b cel3623; c cel36

Article Snippet: The nucleotide sequence encoding the twin-arginine signal peptide of B . subtilis YwbN [ ] and codon-optimized cel7482 was chemically synthesized by BGI Inc., Beijing, China.

Techniques: Recombinant, Purification, Incubation, Activity Assay

Journal: Biotechnology for Biofuels

Article Title: Discovery of new cellulases from the metagenome by a metagenomics-guided strategy

doi: 10.1186/s13068-016-0557-3

Figure Lengend Snippet: a The effect of NaCl on the activity of the recombinant cel7482, cel3623 and cel36. Each purified enzyme was incubated with CMC at optimal temperature for 30 min in 50 mM citrate–phosphate buffer (pH 5.5) in the presence of 0.5–5 M NaCl. b Halotolerance of the recombinant cel7482. After pre-incubation in 0.5, 2 and 5 M NaCl for different periods of time, the residual activity was determined at 70 °C in 50 mM citrate–phosphate buffer (pH 5.5) with CMC as substrate

Article Snippet: The nucleotide sequence encoding the twin-arginine signal peptide of B . subtilis YwbN [ ] and codon-optimized cel7482 was chemically synthesized by BGI Inc., Beijing, China.

Techniques: Activity Assay, Recombinant, Purification, Incubation

Journal: Biotechnology for Biofuels

Article Title: Discovery of new cellulases from the metagenome by a metagenomics-guided strategy

doi: 10.1186/s13068-016-0557-3

Figure Lengend Snippet: a The predicted 3D structure of cel7482 ( side view and top view ) using the crystal structure of a family 5 endoglucanase (PDB: 1ceo) as modeling template. The colored sticks represent the conserved residues in the active site. The orange sticks represent the different residues between cel7482 and cel3623. b The predicted 3D structure of cel36 ( side view and top view ) using the crystal structure of endo-1,4-β-glucanase from Bacillus subtilis 168 (PDB: 3pzt) as modeling template. The colored sticks represent the conserved residues in the active site

Article Snippet: The nucleotide sequence encoding the twin-arginine signal peptide of B . subtilis YwbN [ ] and codon-optimized cel7482 was chemically synthesized by BGI Inc., Beijing, China.

Techniques:

Journal: Biotechnology for Biofuels

Article Title: Discovery of new cellulases from the metagenome by a metagenomics-guided strategy

doi: 10.1186/s13068-016-0557-3

Figure Lengend Snippet: Cellulase activities of the culture supernatant and total cell lysate derived from B. subtilis 168 and its tat mutant strains. 168, B. subtilis 168; tatAyCy , tat mutant strain lacking functional TatAyCy translocase; tatAdCd , tat mutant strain lacking functional TatAdCd translocase; total- tat 2 , tat mutant strain lacking all Tat translocases. For expression of YwbN-cel7482 fusion protein, the culture of the recombinant B. subtilis was induction with 0.5 % xylose at 37 °C for 24 h

Article Snippet: The nucleotide sequence encoding the twin-arginine signal peptide of B . subtilis YwbN [ ] and codon-optimized cel7482 was chemically synthesized by BGI Inc., Beijing, China.

Techniques: Derivative Assay, Mutagenesis, Functional Assay, Expressing, Recombinant

Journal: Biotechnology for Biofuels

Article Title: Discovery of new cellulases from the metagenome by a metagenomics-guided strategy

doi: 10.1186/s13068-016-0557-3

Figure Lengend Snippet: Strains, plasmids and primers used in this study

Article Snippet: The nucleotide sequence encoding the twin-arginine signal peptide of B . subtilis YwbN [ ] and codon-optimized cel7482 was chemically synthesized by BGI Inc., Beijing, China.

Techniques: Plasmid Preparation, TA Cloning, Sequencing, Expressing, Recombinant