Abstract
Here, we report the draft genome sequence of Corynebacterium variabile Mu292, which was originally isolated from the surface of Munster, a French smear-ripened cheese. This genome investigation will improve our knowledge on the molecular determinants potentially involved in the adaptation of this strain during the Munster-type cheese manufacturing process.
GENOME ANNOUNCEMENT
Smear-ripened cheeses harbor complex microbial consortia that are mainly responsible for the production of typical sensory properties (1). Their activities are influenced by the technological processes and manufacturing environment. Corynebacterium species are commonly involved in the cheese-ripening process (2–4) and contribute to the flavor and texture of the final product. Three sequenced genomes are currently available for cheese isolates belonging to the Corynebacterium genus. Two are affiliated with Corynebacterium casei and were isolated from a French (5) and an Irish smear-ripened cheese (6), respectively. The third one is affiliated with Corynebacterium variabile and was isolated from Gubbeen (7).
We report here the genome sequence of Corynebacterium variabile Mu292, isolated in 1989 from Munster, a soft smear-ripened cheese. Sequencing was performed using Illumina MiSeq technology. After filtering, a total of 1,169,642 paired-ends reads of 250 bp in length were generated and merged using FLASH (8). De novo assembly was performed using SPAdes (version 3.1.1, with default parameters) (9), which generated 66 large contigs (≥1,000 bp), with an average sequencing coverage of 100-fold. The unclosed draft genome is 3,185,550 bp in length and has a G+C content of 67.3%. Gene prediction and annotation were performed using the IMG system, as described previously (10). This genome encompasses 3,007 genes, including 2,942 coding DNA sequences, 7 rRNAs, and 58 tRNAs.
Comparative analysis of the genome of C. variabile Mu292 with the genome of C. variabile strain DSM 44702, isolated from Gubbeen cheese (7), will provide valuable insights into the adaptation of C. variabile strains to different cheese technologies. Indeed, Gubbeen and Munster cheeses are differentiated by their technological characters, such as pH of the curd and NaCl and dry-matter contents (11, 12). Interestingly, the presence of a type I restriction-modification system in the genome of C. variabile Mu292 might explain why it is devoid of the phage-related chromosomal island of C. variabile DSM 44702 (7, 13).
Another feature in the genome of C. variabile Mu292 is the presence of a gene coding for a putative arylsulfatase (EC 3.1.6.1), sharing 82% sequence identity (protein level) with the sequence of Corynebacterium terpenotabidum Y-11T (NCBI accession no. WP_020440046), a bacterium isolated from soil and which is phylogenetically close to C. variabile (14, 15). This enzyme has been previously described in various soil bacteria and is considered as a key enzyme in sulfur metabolism (16, 17). In the cheese habitat, arylsulfatase may be involved in the release of molecules conjugated with sulfate, such as alkylphenols, which contribute to sheep-like flavors of the cheeses manufactured from sheep’s milk (18). Thus, this specificity found in the genome of C. variabile Mu292 might be of interest for understanding sulfur metabolism in cheese, which is of great importance for the cheese-making process (19).
This second genome sequence of Corynebacterium variabile will allow deeper comparative genomic studies among Corynebacterium species and other Actinobacteria, provides new elements for understanding the adaptation strategies of cheese bacteria to the cheese habitat, and potentially aids in discovering novel technological properties for the food industry.
Nucleotide sequence accession numbers.
The draft genome sequences of Corynebacterium variabile Mu292 have been deposited at the EMBL database under accession numbers FAUH01000001 to FAUH01000066.
ACKNOWLEDGMENTS
We thank the INRA GeT-PlaGE platform (http://get.genotoul.fr) for sequencing the genome, the INRA MIGALE bioinformatics platform (http://migale.jouy.inra.fr) for providing computational resources and support, and the IMG-ER pipeline for generating annotations.
Footnotes
Citation Dugat-Bony E, Sarthou A-S, Loux V, Vidal M, Bonnarme P, Irlinger F, Layec S. 2016. Draft genome sequence of Corynebacterium variabile Mu292, isolated from Munster, a French smear-ripened cheese. Genome Announc 4(4):e00669-16. doi:10.1128/genomeA.00669-16.
REFERENCES
- 1.Irlinger F, Layec S, Hélinck S, Dugat-Bony E. 2015. Cheese rind microbial communities: diversity, composition and origin. FEMS Microbiol Lett 362:1–11. doi: 10.1093/femsle/fnu015. [DOI] [PubMed] [Google Scholar]
- 2.Feurer C, Irlinger F, Spinnler HE, Glaser P, Vallaeys T. 2004. Assessment of the rind microbial diversity in a farmhouse-produced vs a pasteurized industrially produced soft red-smear cheese using both cultivation and rDNA-based methods. J Appl Microbiol 97:546–556. doi: 10.1111/j.1365-2672.2004.02333.x. [DOI] [PubMed] [Google Scholar]
- 3.Brennan NM, Ward AC, Beresford TP, Fox PF, Goodfellow M, Cogan TM. 2002. Biodiversity of the bacterial flora on the surface of a smear cheese. Appl Environ Microbiol 68:820–830. doi: 10.1128/AEM.68.2.820-830.2002. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 4.Monnet C, Correia K, Sarthou A-S, Irlinger F. 2006. Quantitative detection of Corynebacterium casei in cheese by real-time PCR. Appl Environ Microbiol 72:6972–6979. doi: 10.1128/AEM.01303-06. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 5.Monnet C, Loux V, Bento P, Gibrat J-F, Straub C, Bonnarme P, Landaud S, Irlinger F. 2012. Genome sequence of Corynebacterium casei UCMA 3821, isolated from a smear-ripened cheese. J Bacteriol 194:738–739. doi: 10.1128/JB.06496-11. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 6.Walter F, Albersmeier A, Kalinowski J, Rückert C. 2014. Complete genome sequence of Corynebacterium casei LMG S-19264T (=DSM 44701T), isolated from a smear-ripened cheese. J Biotechnol 189:76–77. doi: 10.1016/j.jbiotec.2014.08.038. [DOI] [PubMed] [Google Scholar]
- 7.Schröder J, Maus I, Trost E, Tauch A. 2011. Complete genome sequence of Corynebacterium variabile DSM 44702 isolated from the surface of smear-ripened cheeses and insights into cheese ripening and flavor generation. BMC Genomics 12:545. doi: 10.1186/1471-2164-12-545. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 8.Magoč T, Salzberg SL. 2011. FLASH: fast length adjustment of short reads to improve genome assemblies. Bioinformatics 27:2957–2963. doi: 10.1093/bioinformatics/btr507. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 9.Bankevich A, Nurk S, Antipov D, Gurevich AA, Dvorkin M, Kulikov AS, Lesin VM, Nikolenko SI, Pham S, Prjibelski AD, Pyshkin AV, Sirotkin AV, Vyahhi N, Tesler G, Alekseyev MA, Pevzner PA. 2012. SPAdes: a new genome assembly algorithm and its applications to single-cell sequencing. J Comput Biol 19:455–477. doi: 10.1089/cmb.2012.0021. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 10.Huntemann M, Ivanova NN, Mavromatis K, Tripp HJ, Paez-Espino D, Palaniappan K, Szeto E, Pillay M, Chen I-MA, Pati A, Nielsen T, Markowitz VM, Kyrpides NC. 2015. The standard operating procedure of the DOE-JGI microbial genome annotation pipeline (MGAP v.4). Stand Genomic Sci 10:86. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 11.Cogan TM, Goerges S, Gelsomino R, Larpin S, Hohenegger M, Bora N, Jamet E, Rea MC, Mounier J, Vancanneyt M, Guéguen M, Desmasures N, Swings J, Goodfellow M, Ward AC, Sebastiani H, Irlinger F, Chamba J-F, Beduhn R, Scherer S. 2014. Biodiversity of the surface microbial consortia from Limburger, Reblochon, Livarot, Tilsit, and Gubbeen cheeses. Microbiol Spectr 2:CM-0010-2012. doi: 10.1128/microbiolspec.CM-0010-2012. [DOI] [PubMed] [Google Scholar]
- 12.Leclercq-Perlat M-N, Spinnler H-E. 2010. The type of cheese curds determined the colouring capacity of Brevibacterium and Arthrobacter species. J Dairy Res 77:287–294. doi: 10.1017/S0022029910000245. [DOI] [PubMed] [Google Scholar]
- 13.Loenen WAM, Dryden DTF, Raleigh EA, Wilson GG. 2014. Type I restriction enzymes and their relatives. Nucleic Acids Res 42:20–44. doi: 10.1093/nar/gkt847. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 14.Rückert C, Albersmeier A, Al-Dilaimi A, Bednarz H, Niehaus K, Szczepanowski R, Kalinowski J. 2014. Genome sequence of the squalene-degrading bacterium Corynebacterium terpenotabidum type strain Y-11T (=DSM 44721T). Stand Genomic Sci 9:505–513. doi: 10.4056/sigs.4588337. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 15.Takeuchi M, Sakane T, Nihira T, Yamada Y, Imai K. 1999. Corynebacterium terpenotabidum sp. nov., a bacterium capable of degrading squalene. Int J Syst Bacteriol 49:223–229. doi: 10.1099/00207713-49-1-223. [DOI] [PubMed] [Google Scholar]
- 16.Cregut M, Piutti S, Vong P-C, Slezack-Deschaumes S, Crovisier I, Benizri E. 2009. Density, structure, and diversity of the cultivable arylsulfatase-producing bacterial community in the rhizosphere of field-grown rape and barley. Soil Biol Biochem 41:704–710. doi: 10.1016/j.soilbio.2009.01.005. [DOI] [Google Scholar]
- 17.Cregut M, Piutti S, Slezack-Deschaumes S, Benizri E. 2013. Compartmentalization and regulation of arylsulfatase activities in Streptomyces sp., Microbacterium sp. and Rhodococcus sp. soil isolates in response to inorganic sulfate limitation. Microbiol Res 168:12–21. doi: 10.1016/j.micres.2012.08.001. [DOI] [PubMed] [Google Scholar]
- 18.Kilic M, Lindsay RC. 2005. Enrichment of cheeses manufactured from cow’s and sheep’s milk blends with sheep-like species-related alkylphenols. J Agric Food Chem 53:1707–1712. doi: 10.1021/jf0484834. [DOI] [PubMed] [Google Scholar]
- 19.Landaud S, Helinck S, Bonnarme P. 2008. Formation of volatile sulfur compounds and metabolism of methionine and other sulfur compounds in fermented food. Appl Microbiol Biotechnol 77:1191–1205. doi: 10.1007/s00253-007-1288-y. [DOI] [PubMed] [Google Scholar]