Abstract
Salinivibrio proteolyticus M318, a halophilic bacterium isolated from fermented shrimp paste, is able to produce polyhydroxyalkanoate (PHA) from different carbon sources. In this study, we report the whole-genome sequence of strain M138, which comprises 2 separated chromosomes and 2 plasmids, and the complete genome contains 3,605,935 bp with an average GC content of 49.9%. The genome of strain M318 contains 3341 genes, 98 tRNA genes, and 28 rRNA genes. The 16S rRNA gene sequence and average nucleotide identity analysis associated with morphological and biochemical tests showed that this strain has high homology to the reference strain Salinivibrio proteolyticus DSM 8285. The genes encoding key enzymes for PHA and ectoine synthesis were identified from the bacterial genome. In addition, the TeaABC transporter responsible for ectoine uptake from the environment and the operon doeABXCD responsible for the degradation of ectoine were also detected. Strain M318 was able to produce poly(3-hydroxybutyrate) [P(3HB)] from different carbon sources such as glycerol, maltose, glucose, fructose, and starch. The ability to produce ectoines at different NaCl concentrations was investigated. High ectoine content of 26.2% of cell dry weight was obtained by this strain at 18% NaCl. This report provides genetic information regarding adaptive mechanisms of strain M318 to stress conditions, as well as new knowledge to facilitate the application of this strain as a bacterial cell factory for the production of PHA and ectoine.
Similar content being viewed by others
References
Amoozegar MA, Schumann P, Hajighasemi M, Fatemi AZ, Karbalaei-Heidari HR (2008) Salinivibrio proteolyticus sp. nov., a moderately halophilic and proteolytic species from a hypersaline lake in Iran. Int J Syst Evol Microbiol 58:1159–1163
Anh DBQ, Mi NTT, Huy DNA, Hung PV (2015) Isolation and optimization of growth condition of Bacillus sp. from fermented shrimp paste for high fibrinolytic enzyme production. Arab J Sci Eng 40:23–28
Chamroensaksri N, Tanasupawat S, Akaracharanya A, Visessanguan W, Kudo T, Itoh T (2009) Salinivibrio siamensis sp. nov., from fermented fish (pla-ra) in Thailand. Int J Syst Evol Microbiol 59:880–885
Czech L, Hermann L, Stöveken N, Richter AA, Höppner A, Smits SHJ, Heider J, Bremer E (2018) Role of the extremolytes ectoine and hydroxyectoine as stress protectants and nutrients: genetics, phylogenomics, biochemistry, and structural analysis. Genes 9:177
de la Haba RR, López-Hermoso C, Sánchez-Porro C, Konstantinidis KT, Ventosa A (2019) Comparative genomics and phylogenomic analysis of the genus Salinivibrio. Front Microbiol 10:2104
Favaro L, Basaglia M, Casella S (2019) Improving polyhydro-xyalkanoate production from inexpensive carbon sources by genetic approaches: a review. Biofuels Bioprod Biorefin 13:208–227
Galisteo C, Sánchez-Porro C, de la Haba RR, López-Hermoso C, Fernández AB, Farias ME, Ventosa A (2019) Characterization of Salinivibrio socompensis sp. nov., a new halophilic bacterium isolated from the high-altitude hypersaline lake Socompa, Argentina. Microorganisms 7:241
Goris J, Konstantinidis KT, Klappenbach JA, Coenye T, Vandamme P, Tiedje JM (2007) DNA-DNA hybridization values and their relationship to whole genome sequence similarities. Int J Syst Evol Microbiol 57:81–91
Grammann K, Volke A, Kunte HJ (2002) New type of osmoregulated solute transporter identified in halophilic members of the bacteria domain: TRAP transporter TeaABC mediates uptake of ectoine and hydroxyectoine in Halomonas elongata DSM 2581T. J Bacteriol 184:3078–3085
Guzmán C, Hurtado A, Carreno C, Casos I (2017) Production of polyhydroxyalkanoates by native halophilic bacteria using Solanum tuberosum L. shell starch. Sci Agropecu 8:109–118
Huijberts GNM, van der Wal H, Wilkinson C, Eggink G (1994) Gas-chromatographic analysis of poly(3-hydroxyalkanoates) in bacteria. Biotechnol Tech 8:187–192
Huu-Phong T, Van-Thuoc D, Sudesh K (2016) Biosynthesis of poly(3-hydroxybutyrate) and its copolymer by Yangia sp. ND199 from different carbon sources. Int J Biol Macromol 84:361–266
Kunte HJ, Galinski EA, Trüper HG (1993) A modified FMOC-method for the detection of aminoacid type osmolytes and tetrahy-dropyrimidines (ectoines). J Microbiol Methods 17:129–136
Kunte HJ, Lentzen G, Galinski EA (2014) Industrial production of the cell protectant ectoine: production mechanisms, processes, and products. Curr Biotechnol 3:10–25
Lee I, Kim YO, Park SC, Chun J (2016) OrthoANI: an improved algorithm and software for calculating average nucleotide identity. Int J Syst Evol Microbiol 66:1100–1103
Lentzen G, Schwarz T (2006) Extremolytes: natural compounds from extremophiles for versatile applications. Appl Microbiol Biotechnol 72:623–634
Lippert G, Galinski EA (1992) Enzyme stabilization by ectoine-type compatible solutes: protection against heating, freezing and drying. Appl Microbiol Biotechnol 37:61–65
López-Hermoso C, de la Haba RR, Sánchez-Porro C, Ventosa A (2018a) Emended description of Salinivibrio proteolyticus, including Salinivibrio costicola subsp. vallismortis and five new isolates. Int J Syst Evol Microbiol 68:1599–1607
López-Hermoso C, de la Haba RR, Sánchez-Porro C, Ventosa A (2018b) Salinivibrio kushneri sp. nov., a moderately halophilic bacteriumisolated from salterns. Syst Appl Microbiol 41:159–166
Mellado E, Moore ERB, Nieto JJ, Ventosa A (1996) Analysis of 16S rRNA gene sequences of Vibrio costicola strains: description of Salinivibrio costicola gen. nov., comb. nov. Int J Syst Bacteriol 46:817–821
Mohandas SP, Balan L, Lekshmi N, Cubelio SS, Philip R, Bright Singh IS (2016) Production and characterization of polyhydroxybutyrate from Vibrio harveyi MCCB 284 utilizing glycerol as carbon source. J Appl Microbiol 122:698–707
Müller D, Lindemann T, Shah-Hosseini K, Scherner O, Hnop M, Bilstein A, Mösges R (2016) Efficacy and tolerability of an ectoine mouth and throat spray compared with those of saline lozenges in the treatment of acute pharyngitis and/or laryngitis: a perspective, controlled, observational clinical trial. Eur Arch Otorhinolaryngol 273:2591–2597
Obruca S, Sedlacek P, Slaninova E, Fritz I, Daffert C, Meixner K, Sedrlova Z, Koller M (2020) Novel unexpected functions of PHA gralunes. Appl Microbiol Biotechnol 104:4795–4810
Oren A (2008) Microbial life at high salt concentrations: phylogenetic and metabolic diversity. Saline Systems 4:2
Pongsetkul J, Benjakul S, Vongkamjan K, Sumpavapol P, Osaka K (2017) Microbiological and chemical changes of shrimp Acetes vulgaris during Kapi production. J Food Sci Technol 54:3473–3482
Poolman B, Glaasker E (1998) Regulation of compatible solute accumulation in bacteria. Mol Microbiol 29:397–407
Quillaguamán J, Guzmán H, Van-Thuoc D, Hatti-Kaul R (2010) Synthesis and production of polyhydroxyalkanoates by halophiles: current potential and future prospects. Appl Microbiol Biotechnol 85:1687–1696
Raza ZA, Abid S, Banat IM (2018) Polyhydroxyalkanoates: characteristics, production, recent developments and applications. Int Biodeterior Biodegradation 126:45–56
Roberts MF (2005) Organic compatible solutes of halotolerant and halophilic microorganism. Saline Systems 1:5
Romano I, Gambacorta A, Lama L, Nicolaus B, Giordano A (2005) Salinivibrio costicola subsp. alcaliphilus subsp. nov., a halo-alkaliphilic aerobe from Campania Region (Italy). Syst Appl Microbiol 28:34–42
Romano I, Orlando P, Gambacorta A, Nicolaus B, Dipasquale L, Pascual J, Giordano A, Lama L (2011) Salinivibrio sharmensis sp. nov., a novel haloalkaliphilic bacterium from a saline lake in Ras Mohammed Park (Egypt). Extremophiles 15:213–220
Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425
Sauer T, Galinski EA (1998) Bacterial milking: a novel bioprocess for production of compatible solutes. Biotechnol Bioeng 57:306–313
Schwibbert K, Marin-Sanguino A, Bagyan I, Heidrich G, Lentzen G, Seitz H, Rampp M, Schuster SC, Klenk HP, Pfeiffer F, Oesterhelt D, Kunte HJ (2011) A blueprint of ectoine metabolism from the genome of the industrial producer Halomonas elongata DSM 2581T. Environ Microbiol 13:1973–1994
Sudesh K, Abe H, Doi Y (2000) Synthesis, structure and properties of polyhydroxyalkanoates: biological polyesters. Prog Polym Sci 25:1503–1555
Tamura K, Stecher G, Peterson D, Filipski A (2013) MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30:2725–2729
Tan D, Xue YS, Aibaidula G, Chen GQ (2011) Unsterile and continuous production of polyhydroxybutyrate by Halomonas TD01. Bioresour Technol 102:8130–8136
Tatusova T, DiCuccio M, Badretdin A, Chetvernin V, Nawrocki EP, Zaslavsky L, Lomsadze A, Pruitt KD, Borodovsky M, Ostell J (2016) NCBI prokaryotic genome annotation pipeline. Nucleic Acids Res 44:6614–6624
Thuoc DV, Hien TT, Sudesh K (2019a) Identification and characterization of ectoine-producing bacteria isolated from Can Gio mangrove soil in Vietnam. Ann Microbiol 69:819–828
Thuoc DV, My DN, Loan TT, Sudesh K (2019b) Utilization of waste fish oil and glycerol as carbon sources for polyhydroxyalkanoate (PHA) production by Salinivibrio sp. M318. Int J Biol Macromol 141:885–892
Van-Thuoc D, Guzmán H, Quillaguamán J, Hatti-Kaul R (2010) High productivity of ectoines by Halomonas boliviensis using a combined two-step fed-batch culture and milking process. J Biotechnol 147:46–51
Funding
This research was supported by the Vietnam National Foundation for Science and Technology Development (Nafosted) (Grant 106-NN.04-2016.11).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of Interest
The authors declare that they have no conflict of interest.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic Supplementary Material
ESM 1
(DOCX 269 kb)
Rights and permissions
About this article
Cite this article
Van Thuoc, D., Loan, T.T., Trung, T.A. et al. Genome Mining Reveals the Biosynthetic Pathways of Polyhydroxyalkanoate and Ectoines of the Halophilic Strain Salinivibrio proteolyticus M318 Isolated from Fermented Shrimp Paste. Mar Biotechnol 22, 651–660 (2020). https://doi.org/10.1007/s10126-020-09986-z
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10126-020-09986-z