Genome Structure of Bacillus cereus tsu1 and Genes Involved in Cellulose Degradation and Poly-3-Hydroxybutyrate Synthesis
In previous work, we reported on the isolation and genome sequence analysis of Bacillus cereus strain tsu1 NCBI accession number JPYN00000000. The 36 scaffolds in the assembled tsu1 genome were all aligned with B. cereus B4264 genome with variations. Genes encoding for xylanase and cellulase and the...
Saved in:
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2017-01-01
|
| Series: | International Journal of Polymer Science |
| Online Access: | http://dx.doi.org/10.1155/2017/6192924 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832549121060241408 |
|---|---|
| author | Hui Li Suping Zhou Terrance Johnson Koen Vercruysse Ouyang Lizhi Parthasarathy Ranganathan Nsoki Phambu Alexander J. Ropelewski Theodore W. Thannhauser |
| author_facet | Hui Li Suping Zhou Terrance Johnson Koen Vercruysse Ouyang Lizhi Parthasarathy Ranganathan Nsoki Phambu Alexander J. Ropelewski Theodore W. Thannhauser |
| author_sort | Hui Li |
| collection | DOAJ |
| description | In previous work, we reported on the isolation and genome sequence analysis of Bacillus cereus strain tsu1 NCBI accession number JPYN00000000. The 36 scaffolds in the assembled tsu1 genome were all aligned with B. cereus B4264 genome with variations. Genes encoding for xylanase and cellulase and the cluster of genes in the poly-3-hydroxybutyrate (PHB) biosynthesis pathway were identified in tsu1 genome. The PHB accumulation in B. cereus tsu1 was initially identified using Sudan Black staining and then confirmed using high-performance liquid chromatography. Physical properties of these PHB extracts, when analyzed with Raman spectra and Fourier transform infrared spectroscopy, were found to be comparable to the standard compound. The five PHB genes in tsu1 (phaA, phaB, phaR, phaC, and phaP) were cloned and expressed with TOPO cloning, and the recombinant proteins were validated using peptide mapping of in-gel trypsin digestion followed by mass spectrometry analysis. The recombinant E. coli BL21 (DE3) (over)expressing phaC was found to accumulate PHB particles. The cellulolytic activity of tsu1 was detected using carboxymethylcellulose (CMC) plate Congo red assay and the shift towards low-molecular size forms of CMC revealed by gel permeation chromatography in CMC liquid culture and the identification of a cellulase in the secreted proteome. |
| format | Article |
| id | doaj-art-b8513440438245298523913f4bfa1072 |
| institution | Kabale University |
| issn | 1687-9422 1687-9430 |
| language | English |
| publishDate | 2017-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | International Journal of Polymer Science |
| spelling | doaj-art-b8513440438245298523913f4bfa10722025-02-03T06:12:16ZengWileyInternational Journal of Polymer Science1687-94221687-94302017-01-01201710.1155/2017/61929246192924Genome Structure of Bacillus cereus tsu1 and Genes Involved in Cellulose Degradation and Poly-3-Hydroxybutyrate SynthesisHui Li0Suping Zhou1Terrance Johnson2Koen Vercruysse3Ouyang Lizhi4Parthasarathy Ranganathan5Nsoki Phambu6Alexander J. Ropelewski7Theodore W. Thannhauser8College of Agricultural, Human and Natural Sciences, Tennessee State University, Nashville, TN, USACollege of Agricultural, Human and Natural Sciences, Tennessee State University, Nashville, TN, USACollege of Agricultural, Human and Natural Sciences, Tennessee State University, Nashville, TN, USACollege of Agricultural, Human and Natural Sciences, Tennessee State University, Nashville, TN, USACollege of Agricultural, Human and Natural Sciences, Tennessee State University, Nashville, TN, USACollege of Agricultural, Human and Natural Sciences, Tennessee State University, Nashville, TN, USACollege of Agricultural, Human and Natural Sciences, Tennessee State University, Nashville, TN, USAPittsburgh Supercomputing Center, Pittsburgh, PA, USAR.W. Holley Center for Agriculture and Health, Plant, Soil and Nutrition Research Unit, USDA, ARS, Ithaca, NY, USAIn previous work, we reported on the isolation and genome sequence analysis of Bacillus cereus strain tsu1 NCBI accession number JPYN00000000. The 36 scaffolds in the assembled tsu1 genome were all aligned with B. cereus B4264 genome with variations. Genes encoding for xylanase and cellulase and the cluster of genes in the poly-3-hydroxybutyrate (PHB) biosynthesis pathway were identified in tsu1 genome. The PHB accumulation in B. cereus tsu1 was initially identified using Sudan Black staining and then confirmed using high-performance liquid chromatography. Physical properties of these PHB extracts, when analyzed with Raman spectra and Fourier transform infrared spectroscopy, were found to be comparable to the standard compound. The five PHB genes in tsu1 (phaA, phaB, phaR, phaC, and phaP) were cloned and expressed with TOPO cloning, and the recombinant proteins were validated using peptide mapping of in-gel trypsin digestion followed by mass spectrometry analysis. The recombinant E. coli BL21 (DE3) (over)expressing phaC was found to accumulate PHB particles. The cellulolytic activity of tsu1 was detected using carboxymethylcellulose (CMC) plate Congo red assay and the shift towards low-molecular size forms of CMC revealed by gel permeation chromatography in CMC liquid culture and the identification of a cellulase in the secreted proteome.http://dx.doi.org/10.1155/2017/6192924 |
| spellingShingle | Hui Li Suping Zhou Terrance Johnson Koen Vercruysse Ouyang Lizhi Parthasarathy Ranganathan Nsoki Phambu Alexander J. Ropelewski Theodore W. Thannhauser Genome Structure of Bacillus cereus tsu1 and Genes Involved in Cellulose Degradation and Poly-3-Hydroxybutyrate Synthesis International Journal of Polymer Science |
| title | Genome Structure of Bacillus cereus tsu1 and Genes Involved in Cellulose Degradation and Poly-3-Hydroxybutyrate Synthesis |
| title_full | Genome Structure of Bacillus cereus tsu1 and Genes Involved in Cellulose Degradation and Poly-3-Hydroxybutyrate Synthesis |
| title_fullStr | Genome Structure of Bacillus cereus tsu1 and Genes Involved in Cellulose Degradation and Poly-3-Hydroxybutyrate Synthesis |
| title_full_unstemmed | Genome Structure of Bacillus cereus tsu1 and Genes Involved in Cellulose Degradation and Poly-3-Hydroxybutyrate Synthesis |
| title_short | Genome Structure of Bacillus cereus tsu1 and Genes Involved in Cellulose Degradation and Poly-3-Hydroxybutyrate Synthesis |
| title_sort | genome structure of bacillus cereus tsu1 and genes involved in cellulose degradation and poly 3 hydroxybutyrate synthesis |
| url | http://dx.doi.org/10.1155/2017/6192924 |
| work_keys_str_mv | AT huili genomestructureofbacilluscereustsu1andgenesinvolvedincellulosedegradationandpoly3hydroxybutyratesynthesis AT supingzhou genomestructureofbacilluscereustsu1andgenesinvolvedincellulosedegradationandpoly3hydroxybutyratesynthesis AT terrancejohnson genomestructureofbacilluscereustsu1andgenesinvolvedincellulosedegradationandpoly3hydroxybutyratesynthesis AT koenvercruysse genomestructureofbacilluscereustsu1andgenesinvolvedincellulosedegradationandpoly3hydroxybutyratesynthesis AT ouyanglizhi genomestructureofbacilluscereustsu1andgenesinvolvedincellulosedegradationandpoly3hydroxybutyratesynthesis AT parthasarathyranganathan genomestructureofbacilluscereustsu1andgenesinvolvedincellulosedegradationandpoly3hydroxybutyratesynthesis AT nsokiphambu genomestructureofbacilluscereustsu1andgenesinvolvedincellulosedegradationandpoly3hydroxybutyratesynthesis AT alexanderjropelewski genomestructureofbacilluscereustsu1andgenesinvolvedincellulosedegradationandpoly3hydroxybutyratesynthesis AT theodorewthannhauser genomestructureofbacilluscereustsu1andgenesinvolvedincellulosedegradationandpoly3hydroxybutyratesynthesis |