Development of the first gene expression system for Salinicoccus strains with potential application in bioremediation of hypersaline wastewaters
Salinicoccus salsiraiae IM408 (=CGMCC13032) is a novel halophilic bacterium that we isolated from the saline soil of Da Gang Oilfield. It tolerates 60 g/l sodium chloride and up to 123 g/l (1.5 M) sodium acetate and has shown a potential application in bioremediation of wastewater with high salt and high chemical oxygen demand (COD). Two plasmids, pS408-1 and pS408-2, were identified in S. salsiraiae IM408, and the sequences and copy numbers of the plasmids were determined. Based on these plasmids, two shuttle vectors containing a replicon for Escherichia coli, ampicillin, and chloramphenicol resistance genes, as well as the replicon from pS408-1 or pS408-2, were constructed and named as pTCS101 and pTCS201, respectively. A suitable host strain, named S. salsiraiae PE01, was also developed from the wild-type by plasmid elimination. Using the plasmid pTCS101 as an expression vector, l-lactate dehydrogenase from Staphylococcus aureus was expressed successfully in S. salsiraiae PE01. This is the first gene expression system for the Salinicoccus genus. It has provided the potential for expression of desired proteins or for establishment of desired pathways in Salinicoccus strains, which would make these halophiles more advantageous in future biotechnological applications.
KeywordsSalinicoccus Plasmid Expression vector Plasmid-free strain Protein expression
We would like to express our gratitude to Dr. Rui Wang (Non-coding RNA and Drug Discovery Key Laboratory of Sichuan Province, Chengdu Medical College) and Dr. Guiming Liu (Technology Transfer Center, Institute of Microbiology, Chinese Academy of Sciences) for their help in sample collection and plasmid characterization during their study in our laboratory.
Compliance with ethical standards
This study was funded by the National Natural Science Foundation of China (grant number 31330001) and the Hundred Talents Program of the Chinese Academy of Sciences (to Hua Xiang).
Conflict of interest
The authors declare that they have no conflict of interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
- Hyun DW, Whon TW, Cho YJ, Chun J, Kim MS, Jung MJ, Shin NR, Kim JY, Kim PS, Yun JH, Lee J, Oh SJ, Bae JW (2013) Genome sequence of the moderately halophilic bacterium Salinicoccus carnicancri type strain CrmT (= DSM 23852T). Stand Genomic Sci 8:255–263. doi: 10.4056/sigs.3967649 CrossRefPubMedPubMedCentralGoogle Scholar
- Morimoto T, Kadoya R, Endo K, Tohata M, Sawada K, Liu S, Ozawa T, Kodama T, Kakeshita H, Kageyama Y, Manabe K, Kanaya S, Ara K, Ozaki K, Ogasawara N (2008) Enhanced recombinant protein productivity by genome reduction in Bacillus subtilis. DNA Res 15:73–81. doi: 10.1093/dnares/dsn002 CrossRefPubMedPubMedCentralGoogle Scholar
- Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory Press, New YorkGoogle Scholar