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      Improved Salt Tolerance and Metabolomics Analysis of Synechococcus elongatus UTEX 2973 by Overexpressing Mrp Antiporters

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          Abstract

          The fast-growing cyanobacterium Synechococcus elongatus UTEX 2973 (Syn2973) is a promising candidate for photosynthetic microbial factory. Seawater utilization is necessary for large-scale cultivation of Syn2973 in the future. However, Syn2973 is sensitive to salt stress, making it necessary to improve its salt tolerance. In this study, 21 exogenous putative transporters were individually overexpressed in Syn2973 to evaluate their effects on salt tolerance. The results showed the overexpression of three Mrp antiporters significantly improved the salt tolerance of Syn2973. Notably, overexpressing the Mrp antiporter from Synechococcus sp. PCC 7002 improved cell growth by 57.7% under 0.4 M NaCl condition. In addition, the metabolomics and biomass composition analyses revealed the possible mechanisms against salt stress in both Syn2973 and the genetically engineered strain. The study provides important engineering strategies to improve salt tolerance of Syn2973 and is valuable for understanding mechanisms of salt tolerance in cyanobacteria.

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          Carbohydrate analysis by a phenol-sulfuric acid method in microplate format.

          Tatsuya Masuko, Akio Minami, Norimasa Iwasaki … (2005)
          Among many colorimetric methods for carbohydrate analysis, the phenol-sulfuric acid method is the easiest and most reliable method. It has been used for measuring neutral sugars in oligosaccharides, proteoglycans, glycoproteins, and glycolipids. This method is used widely because of its sensitivity and simplicity. In its original form, it required 50-450 nmol of monosaccharides or equivalent for analysis and thus is inadequate for precious samples. A scaled-down version requiring only 10-80 nmol of sugars was reported previously. We have now modified and optimized this method to use 96-well microplates for high throughput, to gain greater sensitivity, and to economize the reagents. This modified and optimized method allows longer linear range (1-150 nmol for Man) and excellent sensitivity. Moreover, our method is more convenient, requiring neither shaking nor covering, and takes less than 15 min to complete. The speed and simplicity of this method would make it most suitable for analyses of large numbers of samples such as chromatographic fractions.
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            Phototrophic prokaryotes: the cyanobacteria.

            R Y Stanier, G Cohen-Bazire (1977)
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              Synechococcus elongatus UTEX 2973, a fast growing cyanobacterial chassis for biosynthesis using light and CO2

              Jingjie Yu, Michelle Liberton, Paul Cliften … (2015)
              Photosynthetic microbes are of emerging interest as production organisms in biotechnology because they can grow autotrophically using sunlight, an abundant energy source, and CO2, a greenhouse gas. Important traits for such microbes are fast growth and amenability to genetic manipulation. Here we describe Synechococcus elongatus UTEX 2973, a unicellular cyanobacterium capable of rapid autotrophic growth, comparable to heterotrophic industrial hosts such as yeast. Synechococcus UTEX 2973 can be readily transformed for facile generation of desired knockout and knock-in mutations. Genome sequencing coupled with global proteomics studies revealed that Synechococcus UTEX 2973 is a close relative of the widely studied cyanobacterium Synechococcus elongatus PCC 7942, an organism that grows more than two times slower. A small number of nucleotide changes are the only significant differences between the genomes of these two cyanobacterial strains. Thus, our study has unraveled genetic determinants necessary for rapid growth of cyanobacterial strains of significant industrial potential.
              Article 0 comments Cited 132 times – based on 0 reviews     Review now
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                Author and article information

                Contributors
                Journal
                Journal ID (nlm-ta): Front Bioeng Biotechnol
                Journal ID (iso-abbrev): Front Bioeng Biotechnol
                Journal ID (publisher-id): Front. Bioeng. Biotechnol.
                Title: Frontiers in Bioengineering and Biotechnology
                Publisher: Frontiers Media S.A.
                ISSN (Electronic): 2296-4185
                Publication date (Electronic): 26 May 2020
                Publication date Collection: 2020
                Volume: 8
                Electronic Location Identifier: 500
                Affiliations
                [1] 1Laboratory of Synthetic Microbiology, School of Chemical Engineering and Technology, Tianjin University , Tianjin, China
                [2] 2Key Laboratory of Systems Bioengineering, Frontier Science Center for Synthetic Biology, Ministry of Education of China , Tianjin, China
                [3] 3Collaborative Innovation Center of Chemical Science and Engineering , Tianjin, China
                [4] 4Center for Biosafety Research and Strategy, Tianjin University , Tianjin, China
                Author notes

                Edited by: Zhanglin Lin, South China University of Technology, China

                Reviewed by: Xuefeng Lu, Qingdao Institute of Bioenergy and Bioprocess Technology (CAS), China; Konstantinos Vavitsas, National and Kapodistrian University of Athens, Greece; Anne M. Ruffing, Sandia National Laboratories (SNL), United States

                *Correspondence: Fangzhong Wang fangzhong.wang@ 123456tju.edu.cn

                This article was submitted to Synthetic Biology, a section of the journal Frontiers in Bioengineering and Biotechnology

                Article
                DOI: 10.3389/fbioe.2020.00500
                PMC ID: 7264159
                PubMed ID: 32528943
                SO-VID: 7bb36cfb-4c85-4b71-9257-7a47bdb5c6e0
                Copyright © Copyright © 2020 Cui, Sun, Li, Xie, Song, Wang, Chen and Zhang.
                License:

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                Date received : 18 December 2019
                Date accepted : 29 April 2020
                Page count
                Figures: 4, Tables: 3, Equations: 0, References: 62, Pages: 11, Words: 8398
                Funding
                Funded by: National Natural Science Foundation of China 10.13039/501100001809
                Categories
                Subject: Bioengineering and Biotechnology
                Subject: Original Research

                Keywords: cyanobacteria,mrp antiporter,metabolomic analysis,salt tolerance,tolerance engineering
                Data availability:
                Keywords: cyanobacteria, mrp antiporter, metabolomic analysis, salt tolerance, tolerance engineering

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