Isolation and characterization of Micrococcus lylae strain NCT147 from a human microbiome Background

Abstract

As a part of academic lab in microbiology, human skin microbiome samples were swabbed onto a nutrient agar media. Morphologically distinct colonies were purified using quadrant streaking. The pure culture was preserved using glycerol stocking in triplicates. One of these glycerol stocks was revived and used for further morphological and molecular characterization.

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National College Culture Collection Centre www.ncccc.in
PG & Research Department of Biotechnology and Microbiology
National College, Tiruchirappalli- 620 001, Tamil Nadu, India
1
N4C
Isolation and characterization of Micrococcus lylae strain NCT147 from a human
microbiome
Background
As a part of academic lab in microbiology, human skin microbiome samples were swabbed onto
a nutrient agar media. Morphologically distinct colonies were purified using quadrant streaking.
The pure culture was preserved using glycerol stocking in triplicates. One of these glycerol stocks
was 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.

National College Culture Collection Centre www.ncccc.in
PG & Research Department of Biotechnology and Microbiology
National College, Tiruchirappalli- 620 001, Tamil Nadu, India
2
N4C
Strain description: NCT147 grows stringently in nutrient agar plate at 37°C in 24 hours. Colonies
are small, round, milky white, shiny with smooth edges and swarming (in flanking streak lines in
every quadrant). Cells are Gram positive, non-motile cocci. Biochemical reaction implies Indole
negative, methyl red negative, voges proskauer negative and citrate positive. Triple Sugar Iron
implies weak acid butt and alkaline slant with no gas production. Catalase positive and oxidase
negative. The strain NCT72 was sensitive to rifampicin, penicillin and streptomycin and resistant
to erythromycin.
16S rRNA gene sequencing:
Based on the colony, biochemical and molecular characteristics, the strain NCT147 was identified
as Micrococcus lylae. The 16S rRNA gene
sequences of NCT147 was 98.79%
similar Micrococcus lylae (GenBank
accession number: NR_026200) and can
be globally accessed through genbank
accession number OQ255788.
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 1315 positions in the final dataset.
The strain NCT147 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