Whole genome sequencing of bacillus subtilis Natto3 and cloning and expression of ywtA, a pivotal gene of poly-γ-glutamic acid biosynthesis operon

Pandey Tiwari Deepika, Mishra Chatterjee Poonam, Raval Ritu, Dubey Ashok Kumar

Research output: Contribution to journalArticle

Abstract

Poly-γ-glutamic acid (PGA) is produced via PGA synthetase, a membrane-associated enzyme complex encoded by pgs operon consisting of ywsC and ywtABC genes. In our earlier report, we had studied the unique expression pattern of pgs operon in Bacillus subtilis (B. subtilis) Natto3 strain. This motivated us to perform whole genome sequencing of this strain and analyse the sequence of the genes involved in PGA biosynthesis. B. subtilis Natto3 genome has 3,637,088 bp with 44.08% G-C content and no plasmid. It contains 3913 coding sequences (CDSs) with average gene length of 843.33 bp with 57 CDSs lacking matches. A recent study emphasised the conserved nature of ywtA gene in pgs operon. This gene may provide further insights into the role of individual genes in PGA biosynthesis. In this study, we have successfully cloned the ywtA gene in E. coli DH5α and expressed in BL21 (DE3) strain. The purified protein was ~16 kDa which was in congruence with the existing studies. Protein identification using Nano LC-MS/MS showed homology to CapC protein of B. anthracis with a match of 15%.

Original languageEnglish
Pages (from-to)156-166
Number of pages11
JournalResearch Journal of Biotechnology
Volume14
Issue number7
Publication statusPublished - 01-07-2019

Fingerprint

Cloning
Biosynthesis
Bacilli
Operon
Bacillus subtilis
Organism Cloning
Glutamic Acid
Genes
Genome
Acids
Proteins
Base Composition
Ligases
Sequence Analysis
Plasmids
Escherichia coli
Enzymes
Membranes

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Cite this

@article{c797585c3f0148bb99ff2755ae52a7e4,
title = "Whole genome sequencing of bacillus subtilis Natto3 and cloning and expression of ywtA, a pivotal gene of poly-γ-glutamic acid biosynthesis operon",
abstract = "Poly-γ-glutamic acid (PGA) is produced via PGA synthetase, a membrane-associated enzyme complex encoded by pgs operon consisting of ywsC and ywtABC genes. In our earlier report, we had studied the unique expression pattern of pgs operon in Bacillus subtilis (B. subtilis) Natto3 strain. This motivated us to perform whole genome sequencing of this strain and analyse the sequence of the genes involved in PGA biosynthesis. B. subtilis Natto3 genome has 3,637,088 bp with 44.08{\%} G-C content and no plasmid. It contains 3913 coding sequences (CDSs) with average gene length of 843.33 bp with 57 CDSs lacking matches. A recent study emphasised the conserved nature of ywtA gene in pgs operon. This gene may provide further insights into the role of individual genes in PGA biosynthesis. In this study, we have successfully cloned the ywtA gene in E. coli DH5α and expressed in BL21 (DE3) strain. The purified protein was ~16 kDa which was in congruence with the existing studies. Protein identification using Nano LC-MS/MS showed homology to CapC protein of B. anthracis with a match of 15{\%}.",
author = "Deepika, {Pandey Tiwari} and Poonam, {Mishra Chatterjee} and Raval Ritu and Kumar, {Dubey Ashok}",
year = "2019",
month = "7",
day = "1",
language = "English",
volume = "14",
pages = "156--166",
journal = "Research Journal of Biotechnology",
issn = "0973-6263",
publisher = "Research Journal of BioTechnology",
number = "7",

}

Whole genome sequencing of bacillus subtilis Natto3 and cloning and expression of ywtA, a pivotal gene of poly-γ-glutamic acid biosynthesis operon. / Deepika, Pandey Tiwari; Poonam, Mishra Chatterjee; Ritu, Raval; Kumar, Dubey Ashok.

In: Research Journal of Biotechnology, Vol. 14, No. 7, 01.07.2019, p. 156-166.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Whole genome sequencing of bacillus subtilis Natto3 and cloning and expression of ywtA, a pivotal gene of poly-γ-glutamic acid biosynthesis operon

AU - Deepika, Pandey Tiwari

AU - Poonam, Mishra Chatterjee

AU - Ritu, Raval

AU - Kumar, Dubey Ashok

PY - 2019/7/1

Y1 - 2019/7/1

N2 - Poly-γ-glutamic acid (PGA) is produced via PGA synthetase, a membrane-associated enzyme complex encoded by pgs operon consisting of ywsC and ywtABC genes. In our earlier report, we had studied the unique expression pattern of pgs operon in Bacillus subtilis (B. subtilis) Natto3 strain. This motivated us to perform whole genome sequencing of this strain and analyse the sequence of the genes involved in PGA biosynthesis. B. subtilis Natto3 genome has 3,637,088 bp with 44.08% G-C content and no plasmid. It contains 3913 coding sequences (CDSs) with average gene length of 843.33 bp with 57 CDSs lacking matches. A recent study emphasised the conserved nature of ywtA gene in pgs operon. This gene may provide further insights into the role of individual genes in PGA biosynthesis. In this study, we have successfully cloned the ywtA gene in E. coli DH5α and expressed in BL21 (DE3) strain. The purified protein was ~16 kDa which was in congruence with the existing studies. Protein identification using Nano LC-MS/MS showed homology to CapC protein of B. anthracis with a match of 15%.

AB - Poly-γ-glutamic acid (PGA) is produced via PGA synthetase, a membrane-associated enzyme complex encoded by pgs operon consisting of ywsC and ywtABC genes. In our earlier report, we had studied the unique expression pattern of pgs operon in Bacillus subtilis (B. subtilis) Natto3 strain. This motivated us to perform whole genome sequencing of this strain and analyse the sequence of the genes involved in PGA biosynthesis. B. subtilis Natto3 genome has 3,637,088 bp with 44.08% G-C content and no plasmid. It contains 3913 coding sequences (CDSs) with average gene length of 843.33 bp with 57 CDSs lacking matches. A recent study emphasised the conserved nature of ywtA gene in pgs operon. This gene may provide further insights into the role of individual genes in PGA biosynthesis. In this study, we have successfully cloned the ywtA gene in E. coli DH5α and expressed in BL21 (DE3) strain. The purified protein was ~16 kDa which was in congruence with the existing studies. Protein identification using Nano LC-MS/MS showed homology to CapC protein of B. anthracis with a match of 15%.

UR - http://www.scopus.com/inward/record.url?scp=85069515366&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85069515366&partnerID=8YFLogxK

M3 - Article

AN - SCOPUS:85069515366

VL - 14

SP - 156

EP - 166

JO - Research Journal of Biotechnology

JF - Research Journal of Biotechnology

SN - 0973-6263

IS - 7

ER -