Sharpea azabuensis KH1P5

S. azabuensis KH1P5 in liquid culture exhibiting its dense layer of EPS. Source: Dr. Roy Rodriguez Carrero

Introduction

Sharpea azabuensis KH1P5 is a high-interest microbe to the Global Methane Hub. It produces a thick, mucoidal layer of extracellular polymeric substance (EPS) that requires removal via washing with chelating agents and detergent before work with the culture.

Global Methane Hub Metadata

Origin:	New Zealand, Kelly Lab
Sequenced by:	Sam Mahoney Kurpe
Assembled by:	William G. Alexander
Methylome called by:	William G. Alexander
Methyltransferases cloned by:	Roy Rodriguez Carrero
MIC and Culturing Conditions by:	Roy Rodriguez Carrero
Transformation by:	Roy Rodriguez Carrero

Genome

Circos plot of S. azabuensis KH1P5. Source: Dr. William G. Alexander

Methylome

While S. azabuensis KH1P5 possesses two putative Type II restriction endonucleases, no methylation is detectable in the isolate. Also, these two genes do not methylate in E. coli strains, suggesting that they're vestigial defense systems.

Methylating E. coli Strains

Use a strain of E. coli that lacks dam, dcm, and hsdRMS, such as AG5589.

Culturing Conditions

S. azabuensis KH1P5 grows well in stock RCM at 37 °C.

Antibiotic MIC Tests

MIC tests of S. azabuensis. Source: Dr. Roy Rodriguez Carrero

antibiotic	concentration (ug/mL)
chloramphenicol	0.38
erythromycin	1
kanamycin	insensitive
spectinomycin	96
tetracycline	0.23

Protocols

• Removal of extracellular material via chelation washing