RESEARCH ARTICLE

Complete genome sequence of Bacillus coagulans CACC834 isolated from canine

Jung-Ae Kim1,2https://orcid.org/0000-0002-0694-477X, Dae-Hyuk Kim1,3https://orcid.org/0000-0002-9948-5313, Yangseon Kim1,*https://orcid.org/0000-0002-8285-3407
Author Information & Copyright
1Department of Research and Development, Center for Industrialization of Agricultural and Livestock Microorganisms, Jeongeup 56212, Korea
2Department of Bioactive Material Science, Jeonbuk National University, Jeonju 54896, Korea
3Department of Molecular Biology, Department of Bioactive Material Science, Institute for Molecular Biology and Genetics, Jeonbuk National University, Jeonju 54896, Korea
*Corresponding author: Yangseon Kim, Department of Research and Development, Center for Industrialization of Agricultural and Livestock Microorganisms, Jeongeup 56212, Korea., Tel: +82-63-536-6712, E-mail: yangseon@cialm.or.kr

© Copyright 2021 Korean Society of Animal Science and Technology. This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Received: Jun 21, 2021; Revised: Sep 15, 2021; Accepted: Sep 25, 2021

Published Online: Nov 30, 2021

Abstract

Bacillus coagulans CACC 834 was isolated from canine feces, and its potential probiotic properties were characterized by functional genome analysis. Whole-genome sequencing of B. coagulans CACC 834 was performed using the PacBio RSII platforms. The complete genome assembly consisted of one circular chromosome (3.1 Mb) with guanine (G) + cytosine (C) content of 47.1%. Annotation revealed 3,181 protein-coding sequences (CDSs), 30 rRNAs, and 83 tRNAs. Gene associated 11% of the genes were involved in replication, recombination, and repair. We also annotated various stress-related, acid resistance, bile salt resistance and adhesion-related domains in this strain, which likely provide support in exerting probiotic action by survival under gastrointestinal tract. These results add to our comprehensive understanding of B. coagulans and suggest potential mammal-related industrial applications.

Keywords: Bacillus coagulans; Canine; Whole-genome sequencing

Bacillus coagulans strains are gram-positive, spore-forming, and produce lactic acid, which possesses the capacity to balance intestinal gut microbiota, ultimately promoting the growth of animals and improving immunity [1,2]. In addition, B. coagulans is resistant to high temperatures because of its probiotic activity, modulates and strengthens the immune system to protect against infections, and minimizes inflammation-related tissue damage [3].

We isolated B. coagulans CACC 834 (KACC 22145) from the feces of a female 3-year-old Boston terrier in Korea. The samples were incubated under anaerobic conditions at 37°C [4].

Whole-genome sequencing was performed using the Pacific Biosciences (PacBio) RS II Single Molecule Real Time (SMRT) platform with a 20 kb SMRTbellTM template library at ChunLab. The reads were assembled using the PacBio SMAR Analysis 2.3.0. [5].

The sequences were annotated using the combined results of the automatic National Center for Biotechnology Information (NCBI) Prokaryotic Genome Annotation Pipeline (PGAP) and Rapid Annotations using Subsystems Technology (RAST) [6].

The complete genome of B. coagulans CACC 834 was composed of a 3,077,319 bp circular chromosome with 47.1% guanine (G) + cytosine (C) content. The genome contained 3,181 protein-coding sequences (CDS), 30 rRNAs, and 83 tRNAs (Table 1). The genome features of B. coagulans CACC 834 are summarized in Fig. 1. Among these CDS, 2,909 genes were classified into 20 clusters of orthologous groups (COG) functional categories (Fig. 1B). More than 41% of genes were involved in transport and metabolism, including amino acid, carbohydrate, inorganic ion, nucleotide, coenzyme, and lipid. Interestingly, 11% of the genes were involved in replication, recombination, and repair (Fig. 1B).

Table 1. Genome overview of Bacillus coagulas CACC834
Feature Values
Genome size (bp) 3,077,319
No. of contigs 1
GC content 47.1%
Protein-coding genes (CDSs) 684
rRNA 30
tRNA 83
plasmid 0
Genbank Acession No. CP076597
Download Excel Table
jast-63-6-1464-g1
Fig. 1. Genome features of Bacillus coagulans CACC834. (A) Circular genome maps of B. coagulans CACC834 chromosome. Circles from the outside to the center denote rRNA and tRNA gene, reverse strand CDS, forward strand CDS, GC skew, and GC content. (B) Genome number of COG categorie. CDS, protein-coding genes; COG, clusters of orthologous group; G, guanine; C, cytosine.
Download Original Figure

The genome of B. coagulans CACC 834 possessed hsp20, hsp60, dnaK, dnaJ and grpE operons, encoding enzymes and proteins for heat shock. Also, B. coagulans CACC 834 has heat shock induced genes, such as clpB, clpP, mcsB, and repair-related genes, such as recA and uvrABC. The expression of these genes is expected to help promote cell recovery from heat shock by limiting damage caused by stress [79]. Furthermore, B. coagulans CACC 834 carried genes known to be involved in lactate synthesis, adhesion, acid resistance, and bile resistance (Table 2).

Table 2. Predicted genes involved in probiotic potency in B. coagulans CACC834
Predicted function B. coagulans CACC834
Predicted genes Start position End position Length (bp)
Lactate synthesis ldh 1,433,259 1,434,242 984
Adhesion protein EpsD 393,741 394,838 1,098
FliD 724,445 726,531 2,088
Acid resistance-related
 Protection or repair of macromolecules dnaK 1,737,678 1,737,546 1,827
dnaJ 1,736,410 1,737,546 1,137
grpE 1,739,540 1,740,202 663
recA 125,046 1,226,113 1,068
uvrABC 765,135 767,135 2,001
clpB 108,312 110,762 2,451
clpP 793,689 794,279 591
mcsB 108,312 110,762 2,451
 Fatty acid synthesis fabF 870,816 871,754 1,242
fabH 871,824 873,065 939
fadD 260,088 261,242 1,155
fabI 903,131 903,910 780
 Acid shock response aspS 1,804,625 180,696 1,773
 Metabolic rearrangements alsD 2,318,874 2,319,623 750
 Gylcine betain ABC transport system opuCC 467,919 468,833 915
Bile salt resistance bass 1,090,675 1,091,145 471
Download Excel Table

This study on complete genome sequence of B. coagulans CACC 834 may increase our understanding of the probiotic effects in host healthcare and extend its potential application as an industrial strain.

The complete genome of B. coagulans strain CACC 834 determined in this study has been deposited in the NCBI GenBank database under accession number CP076597.

Competing interests

No potential conflict of interest relevant to this article was reported.

Funding sources

This research was supported by the Strategic Initiative for Microbiomes in Agriculture and Food grant(no.918002-4) and Ministry of Agriculture, Food and Rural Affairs (no.321094-2), Korea.

Acknowledgements

Not applicable.

Availability of data and material

Upon reasonable request, the datasets of this study can be available from the corresponding author.

Authors’ contributions

Conceptualization: Kim Y.

Data curation: Kim JA.

Formal analysis: Kim JA.

Methodology: Kim JA.

Software: Kim JA.

Validation: Kim JA, Kim DH.

Investigation: Kim Y.

Writing - original draft: Kim Y.

Writing - review & editing: Kim JA, Kim DH, Kim Y.

Ethics approval and consent to participate

This article does not require IRB/IACUC approval because there are no human and animal participants.

REFERENCES

1.

Su F, Xu P. Genomic analysis of thermophilic Bacillus coagulans strains: efficient producers for platform bio-chemicals. Sci Rep. 2014; 4:3926

2.

Zhou Y, Zeng Z, Xu Y, Ying J, Wang B, Majeed M, et al. Application of Bacillus coagulans in animal husbandry and its underlying mechanisms. Animals. 2020; 10:454

3.

Konuray G, Erginkaya Z. Potential use of Bacillus coagulans in the food industry. Foods. 2018; 7:92

4.

Jang HJ, Son S, Kim JA, Jung MY, Choi Y, Kim DH, et al. Characterization and functional test of canine probiotics. Front Microbiol. 2021; 12:625562

5.

Lee JE, Heo S, Kim GB. Complete genome sequence of Streptococcus hyointestinalis B19, a strain producing bacteriocin, isolated from chicken feces. J Anim Sci Technol. 2020; 62:420-2

6.

Aziz RK, Bartels D, Best AA, DeJongh M, Disz T, Edwards RA, et al. The RAST Server: rapid annotations using subsystems technology. BMC Genomics. 2008; 9:75

7.

Palop A, Sala FJ, Condon S. Occurrence of a highly heat-sensitive spore subpopulation of Bacillus coagulans STCC 4522 and its conversion to a more heat-stable form. Appl Environ Microbiol. 1997; 63:2246-51

8.

Guzzo J. Biotechnical applications of small heat shock proteins from bacteria. Int J Biochem Cell Biol. 2012; 44:1698-705

9.

Li P, Tian W, Jiang Z, Liang Z, Wu X, Du B. Genomic characterization and probiotic potency of Bacillus sp. DU-106, a highly effective producer of L-lactic acid isolated from fermented yogurt. Front Microbiol. 2018; 9:2216