Isolation and characterization of Paenibacillus pabuli strain NCT17 from human skin microbiome Background

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 pabuli strain NCT17 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: NCT17 grows well in nutrient agar plate at 37°C in 24 hours. Colonies are
small, white, round with smooth edges. Cells are Gram positive, spore forming, and motile 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 NCT17 was sensitive to streptomycin (13mm), and
resistant to
amoxicillin,
rifampicin,
penicillin and
ampicillin.
Carbohydrate
fermentation
test revealed

National College Culture Collection Centre
PG & Research Department of Biotechnology and Microbiology
National College, Tiruchirappalli- 620 001, Tamil Nadu, India
N4C
organic acid production during the utilization
of arabinose and maltose with gas
production.
16S rRNA gene sequencing:
Based on the colony and biochemical
characteristics, the strain NCT17 was
identified as Paenibacillus pabuli. NCT17 was
99.72% similar to Paenibacillus pabuli
(GenBank accession number: NR_040853).
16S rRNA gene sequence of NCT17 can be
globally accessed through genbank accession
number OQ271418.
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 1416
positions in the final dataset.
The strain NCT17 was deposited in National
College Culture Collection Centre (ncccc) and
are freely available for academic and research
purposes. The strain can be order here
www.ncccc.in