Bacillus cellulosilyticus enzymes

Bacillus cellulosilyticus

Bacillus cellulosilyticus N-4 DSM 2522 (Bcell) was originally isolated from soil in a screen for cellulase producers.   The genome of this organism was sequenced by our group as part of a search for novel, alkaline biomass-degrading enzymes.  The genome of Bcell is a single chromosome of 4.7 M with no plasmids present. The most unusual feature of the genome is the presence of 23 LPXTA membrane anchor proteins, 17 of which are annotated as involved in polysaccharide degradation.  Consistent with the genomic analysis, cell fractionation and enzymatic assay of Bcell cultures show that the majority of polysaccharide degradation is associated with these cell wall LPXTA-enzymes. A BLAST analysis of the LPXTA enzymes of Bcell show they are not closely related to any other enzymes from gram positive organisms, and there is clear evidence that these enzymes are not the result of gene duplication within Bcell, nor were their genes laterally transferred to Bcell from another organism.  These enzymes also possess a number of novel features including multiple, repeated carbohydrate binding modules (CBM) and rarely seen CBM family members including CBM22, CBM23, CBM28, CBM47, and CBM59.

Genomic analysis and growth studies both strongly argue against Bcell being a truly cellulolytic organism, in spite of its name. A more likely  answer is that Bcell degrades fungal mycelia as a preferred carbon source.  This is supported by the large number of annotated chitinases and 1,3 β-glucanases present in the genome.  The organism could utilize these enzymes to degrade fungal cell walls Bcell also possesses two annotated LPXTA adhesion proteins (Bcell_ 0381 and Bcell_2860) as well as several LPXTA hypothetical proteins that may be involved in adhesion to fungi.  We are the only source for these novel Bcell enzymes.  These  enzymes are cloned as soluble, non-LPXTA proteins and produced in E. coli, then purified by column chromatography to give >90% purity by SDS-PAGE.  Enzymes are supplied as solutions in 75 mM Tris-HCl, pH 8.0 and 25% glycerol.

Bell growing on fungal mycelia as carbon source (2000x magnification). Mycelia are dark, thick objects, Bcell cells are thinner and more prevalent.

Bell growing on fungal mycelia as carbon source (2000x magnification). Mycelia are dark, thick objects, Bcell cells are thinner and more prevalent.

 

Bacillus cellulosilyticus enzymes

Name                                    Gene                CAZY Family                      Price

beta-Glucosidase 3      Bcell_0282                  GH1                                   $200/2 mg

beta-Xylosidase 4       Bcell_0385                  GH43                                 $200/2 mg

Cellulase 32                 Bcell_0437                   GH5 CBM5                       $200/2 mg

Cellulase 33                  Bcell_0438                 GH5 CBM5 CBM5           $200/2 mg

Bcell_0537                  Bcell_0537                    GH8                                  $200/2 mg

beta-Xylosidase 2      Bcell_0538                 GH39                                  $200/2 mg            

Xylanase 17                Bcell_0541              (CBM22)3 GH10 CBM9       $200/2 mg

Xylanase 15                Bcell_0547                  GH10                                  $200/2 mg

beta-Xylosidase 5       Bcell_0554                 GH43                                  $200/2 mg

Curdlanase 1               Bcell_0683                 GH16 (CBM4)5                $200/2 mg

Curdlanase 2               Bcell_0690                CBM4 GH16 (CBM4)2     $200/2 mg

beta-Glucosidase 5      Bcell_0705               GH3                                     $200/2 mg

beta-Xylosidase 6        Bcell_0821               GH30                                  $200/2 mg

beta-Galactosidase 4  Bcell_1039              GH43                                   $200/2 mg

beta-Glucosidase 6     Bcell_1042              GH43                                   $200/2 mg

Cellulase 38               Bcell_1191                  GH5 CBM46                       $200/2 mg

beta-Mannanase 7      Bcell_1368         GH26 CBM2 CBM23 CBM59    $200/2 mg

Cellulase 34               Bcell_2363               GH16 CBM6                            $200/2 mg

Galactanase 1            Bcell_2962              CBM61 GH53 CBM47           $200/2 mg

Cellulase 35                Bcell_3371               GH5 CBM17 CBM28            $200/2 mg

Cellulase 36                Bcell_3391                 GH9                                       $200/2 mg

Cellobiohydrolase 3    Bcell_4264            GH3                                        $200/2 mg