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Cite: NAR/gkaa742 2020



PUL General Information

Literature Information

PUL ID

PUL0268

PubMed

23793634, Appl Environ Microbiol. 2013 Sep;79(17):5151-8. doi: 10.1128/AEM.01506-13. Epub 2013 Jun 21.

Characterization method

Northern Blot,promoter assay

Genomic accession number

BA000043.1

Nucelotide position range

726823-732934

Substrate

starch

Loci

GK0704-GK0708

Species

Geobacillus kaustophilus/1462

Degradation or Biosynthesis

degradation

Gene Name

Locus Tag

Protein ID

Gene Position

GenBank Contig Range

EC Number

- GK0704 BAD74989.1 0 - 1275 (+) BA000043.1:726823-728098 -
- GK0705 BAD74990.1 1344 - 2625 (+) BA000043.1:728167-729448 -
- GK0706 BAD74991.1 2624 - 3467 (+) BA000043.1:729447-730290 -
- GK0707 BAD74992.1 3532 - 5074 (+) BA000043.1:730355-731897 3.2.1.1
- GK0708 BAD74993.1 5092 - 6112 (+) BA000043.1:731915-732935 -

Cluster number

1

Gene name

Gene position

Gene type

Found by CGCFinder?

- 1 - 1275 (+) TC: gnl|TC-DB|O07009|3.A.1.1.2 Yes
- 1345 - 2625 (+) TC: gnl|TC-DB|O32261|3.A.1.1.2 Yes
- 2625 - 3467 (+) TC: gnl|TC-DB|O07011|3.A.1.1.2 Yes
- 3533 - 5074 (+) CAZyme: GH13_1|GH13 Yes
- 5093 - 6112 (+) STP: STP|LacI,STP|Peripla_BP_3 No

PUL ID

PUL0268

PubMed

23793634, Appl Environ Microbiol. 2013 Sep;79(17):5151-8. doi: 10.1128/AEM.01506-13. Epub 2013 Jun 21.

Title

Polysaccharide-degrading thermophiles generated by heterologous gene expression in Geobacillus kaustophilus HTA426.

Author

Suzuki H, Yoshida K, Ohshima T

Abstract

Thermophiles have important advantages over mesophiles as host organisms for high-temperature bioprocesses, functional production of thermostable enzymes, and efficient expression of enzymatic activities in vivo. To capitalize on these advantages of thermophiles, we describe here a new inducible gene expression system in the thermophile Geobacillus kaustophilus HTA426. Six promoter regions in the HTA426 genome were identified and analyzed for expression profiles using beta-galactosidase reporter assay. This analysis identified a promoter region upstream of a putative amylose-metabolizing gene cluster that directed high-level expression of the reporter gene. The expression was >280-fold that without a promoter and was further enhanced 12-fold by maltose addition. In association with a multicopy plasmid, this promoter region was used to express heterologous genes. Several genes, including a gene whose product was insoluble when expressed in Escherichia coli, were successfully expressed as soluble proteins, with yields of 0.16 to 59 mg/liter, and conferred new functions to G. kaustophilus strains. Remarkably, cellulase and alpha-amylase genes conferred the ability to degrade cellulose paper and insoluble starch at high temperatures, respectively, generating thermophiles with the potential to degrade plant biomass. Our results demonstrate that this novel expression system expands the potential applications of G. kaustophilus.