A chimeric vacuolar Na+/H+ antiporter gene evolved by DNA family shuffling confers increased salt tolerance in yeast

Wu, Guangxia; Wang, Gang; Ji, Jing; Li, Yong; Gao, Hailing; Jing, Wu; Guan, Wenzhu

doi:10.1016/j.jbiotec.2015.02.033

Cited by 13 publications

(2 citation statements)

References 43 publications

(42 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Further, salt tolerance in yeast was improved using two homologous Na + /H + antiporters from halophytes Salicornia europaea (SeNHX1) and Suaeda salsa (SsNHX1). The mutant exhibited up to 46% salt tolerance compared to parent strain ( Wu et al, 2015 ).…”

Section: Improvement Of the Existing Enzymes Or Proteinsmentioning

confidence: 98%

Current Technological Improvements in Enzymes toward Their Biotechnological Applications

et al. 2016

View full text Add to dashboard Cite

Enzymes from extremophiles are creating interest among researchers due to their unique properties and the enormous power of catalysis at extreme conditions. Since community demands are getting more intensified, therefore, researchers are applying various approaches viz. metagenomics to increase the database of extremophilic species. Furthermore, the innovations are being made in the naturally occurring enzymes utilizing various tools of recombinant DNA technology and protein engineering, which allows redesigning of the enzymes for its better fitment into the process. In this review, we discuss the biochemical constraints of psychrophiles during survival at the lower temperature. We summarize the current knowledge about the sources of such enzymes and their in vitro modification through mutagenesis to explore their biotechnological potential. Finally, we recap the microbial cell surface display to enhance the efficiency of the process in cost effective way.

show abstract

Section: Improvement Of the Existing Enzymes Or Proteinsmentioning

confidence: 98%

Current Technological Improvements in Enzymes toward Their Biotechnological Applications

et al. 2016

View full text Add to dashboard Cite

show abstract

“…A Na + /H + antiporter gene SseNHX1 , which is a chimeric gene of SsNHX1 and SeNHX1 (from halophytes Suaeda salsa and Salicornia europaea , respectively), was obtained by DNA family shuffling. The gene enhanced salt tolerance by 146% and 122% more than the template, respectively [8]. …”

Section: Introductionmentioning

confidence: 99%

Directed Evolution of Dunaliella salina Ds-26-16 and Salt-Tolerant Response in Escherichia coli

Guo

Dong

Xiao

et al. 2016

IJMS

View full text Add to dashboard Cite

Identification and evolution of salt tolerant genes are crucial steps in developing salt tolerant crops or microorganisms using biotechnology. Ds-26-16, a salt tolerant gene that was isolated from Dunaliella salina, encodes a transcription factor that can confer salt tolerance to a number of organisms including Escherichia coli (E. coli), Haematococcus pluvialis and tobacco. To further improve its salt tolerance, a random mutagenesis library was constructed using deoxyinosine triphosphate-mediated error-prone PCR technology, and then screened using an E. coli expression system that is based on its broad-spectrum salt tolerance. Seven variants with enhanced salt tolerance were obtained. Variant EP-5 that contained mutation S32P showed the most improvement with the E. coli transformant enduring salt concentrations up to 1.54 M, in comparison with 1.03 M for the wild type gene. Besides, Ds-26-16 and EP-5 also conferred E. coli transformant tolerance to freezing, cold, heat, Cu2+ and alkaline. Homology modeling revealed that mutation S32P in EP-5 caused the conformational change of N- and C-terminal α-helixes. Expression of Ds-26-16 and EP-5 maintained normal cellular morphology, increased the intracellular antioxidant enzymatic activity, reduced malondialdehyde content, and stimulated Nitric Oxide synthesis, thus enhancing salt tolerance to E. coli transformants.

show abstract

Establishment of a gene function analysis system for the euhalophyte Salicornia europaea L.

Tai

Jiang

et al. 2017

Plant Cell Rep

View full text Add to dashboard Cite

A Salicornia europaea L. in vitro cell transformation system was developed and further applied to SeNHX1 function investigation. The exploration of salt-tolerant genes from halophyte has seriously been limited by the lack of self-dependent transformation system. Here, an Agrobacterium tumefaciens-mediated in vitro cell transformation system of euhalophyte Salicornia europaea L. was developed. Calli derived from hypocotyl of S. europaea were co-cultured for 3 days with Agrobacterium at OD ranging from 1.0 to 1.5 and then selected with 25 mg/L hygromycin (Hyg). The transformed cells were identified from Hyg positive calli by GUS assay and qRT-PCR, and the transformation efficiency was up to 74.4%. The practicality of this system was further tested via genetic manipulation of S. europaea Na/H antiporter 1 (SeNHX1) gene by creating the overexpressing, silencing, and empty vector cells. Survival ratio and Na distribution under salt treatment showed obvious differences in SeNHX1-overexpressing, -silencing, and empty vector cells, indicating the feasibility of this system to analyze gene function. This investigation is enlightening for studies in other non-model plants lacking of self-dependent transformation system.

show abstract

A chimeric vacuolar Na+/H+ antiporter gene evolved by DNA family shuffling confers increased salt tolerance in yeast

Cited by 13 publications

References 43 publications

Current Technological Improvements in Enzymes toward Their Biotechnological Applications

Current Technological Improvements in Enzymes toward Their Biotechnological Applications

Directed Evolution of Dunaliella salina Ds-26-16 and Salt-Tolerant Response in Escherichia coli

Establishment of a gene function analysis system for the euhalophyte Salicornia europaea L.

Contact Info

Product

Resources

About