Isolation and characterization of Paenibacillus phoenicis strain NCT27 from human skin microbiome

Abstract

As a part of academic lab in bacteriology, human skin microbiome was sampled and swabbed on to nutrient agar plates. The pure culture was preserved using glycerol stocking in triplicates. This glycerol stocked cultures were revived and used for further morphological and molecular characterization.

Full text

National College Culture Collection Centre
PG & Research Department of Biotechnology and Microbiology
National College, Tiruchirappalli- 620 001, Tamil Nadu, India
N4C
Isolation and characterization of Paenibacillus phoenicis strain NCT27 from human skin
microbiome
Background
As a part of academic lab in bacteriology, human skin microbiome was sampled and swabbed on
to nutrient agar plates. The pure culture was preserved using glycerol stocking in triplicates. This
glycerol stocked cultures were revived and used for further morphological and molecular
characterization.
Work plan
Bergey’s manual was used to determine the lowest possible taxonomic hierarchy and the strain
was commercially outsourced for bidirectional 16S rRNA gene sequencing for species level
resolution.
Results
Strain description: NCT27 grows well in nutrient agar plate at 37°C in 24 hours. Colonies are
small, greyish white, round with smooth edges. Cells are Gram positive, spore forming, motile

National College Culture Collection Centre
PG & Research Department of Biotechnology and Microbiology
National College, Tiruchirappalli- 620 001, Tamil Nadu, India
N4C
rods. Biochemical reaction implies Indole negative, methyl red positive, voges proskauer negative
and citrate positive. Triple Sugar Iron implies alkaline butt and slant with no gas production.
Catalase positive and oxidase positive. The strain NCT27 was sensitive to streptomycin (18mm),
neomycin (13mm).
16S rRNA gene sequencing:
Based on the colony and biochemical characteristics, the strain NCT27 was identified as
Paenibacillus phoenicis. NCT27 was 99.65% similar to Paenibacillus phoenicis (GenBank
accession number: NR_108292). 16S rRNA gene sequence of NCT27 can be globally accessed
through genbank accession number OQ271426.
The Neighbor-Joining method tree with
percentage of replicate trees in which
the associated taxa clustered together in
the bootstrap test (500 replicates) are
shown. The tree is drawn to scale, with
branch lengths in the same units as
those of the evolutionary distances used
to infer the phylogenetic tree. The
evolutionary distances were computed
using the Maximum Composite
Likelihood method and are in the units
of the number of base substitutions per
site. This analysis involved 11
nucleotide sequences. All ambiguous
positions were removed for each
sequence pair (pairwise deletion
option). There were a total of 1380
positions in the final dataset.
The strain NCT27 was deposited in
National College Culture Collection
Centre (ncccc) and are freely available
for academic and research purposes.
The strain can be obtained here
www.ncccc.in