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Synechocystis sp. PCC 6803
Biotechnology for Biofuels · 2018 ·Vol. 11 ·期 1 ·1-15 · IF 5.497

Adaptive laboratory evolution of cadmium tolerance in Synechocystis sp. PCC 6803

Xu Chunxiao,Sun Tao,Li Shubin,Chen Lei,Zhang Weiwen

Background Cadmium has been a significant threat to environment and human health due to its high toxicity and wide application in fossil-fuel burning and battery industry. Cyanobacteria are one of the most dominant prokaryotes, and the previous studies suggested that they could be valuable in removing Cd2+ from waste water. However, currently, the tolerance to cadmium is very low in cyanobacteria. To further engineer cyanobacteria for the environmental application, it is thus necessary to determine the mechanism that they respond to high concentration of cadmium.Results In this study, a robust strain of Synechocystis PCC 6803 (named ALE-9.0) tolerant to CdSO4 with a concentration up to 9.0 µM was successfully isolated via adaptive laboratory evolution over 802-day continuous passages under cadmium stress. Whole-genome re-sequencing was then performed and nine mutations were identified for the evolved strain compared to the wild-type strain. Among these mutations, a large fragment deletion in slr0454 encoding a cation or drug efflux system protein was found to contribute directly to the resistance to Cd2+ stress. In addition, five other mutations were also demonstrated related to the improved Cd2+ tolerance in ALE-9.0. Moreover, the evolved ALE-9.0 strain was found to obtain cross tolerance to some other heavy metals like zinc and cobalt as well as higher resistance to high light.Conclusion sThe work here identified six genes and their mutations related to Cd2+ tolerance in Synechocystis PCC 6803, and demonstrated the feasibility of adaptive laboratory evolution in tolerance modifications. This work also provided valuable information regarding the cadmium tolerance mechanism in Synechocystis PCC 6803, and useful insights for cyanobacterial robustness and tolerance engineering.

CAS / MDL: 7790-84-3
研究领域: 环境科学 · 微生物学 · 基因组学 · 生物技术
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Frontiers in Microbiology · 2020 · IF 4.236

Light-Driven Biosynthesis of myo-Inositol Directly From CO2 in Synechocystis sp. PCC 6803

Wang Xiaoshuai,Chen Lei,Liu Jing,Sun Tao,Zhang Weiwen

myo-inositol (MI) is an essential growth factor, nutritional source, and important precursor for many derivatives like D-chiro-inositol. In this study, attempts were made to achieve the “green biosynthesis” of MI in a model photosynthetic cyanobacterium Synechocystis sp. PCC 6803. First, several genes encoding myo-inositol-1-phosphate synthases and myo-inositol-1-monophosphatase, catalyzing the first or the second step of MI synthesis, were introduced, respectively, into Synechocystis. The results showed that the engineered strain carrying myo-inositol-1-phosphate synthase gene from Saccharomyces cerevisiae was able to produce MI at 0.97 mg L–1. Second, the combined overexpression of genes related to the two catalyzing processes increased the production up to 1.42 mg L–1. Third, to re-direct more cellular carbon flux into MI synthesis, an inducible small RNA regulatory tool, based on MicC-Hfq, was utilized to control the competing pathways of MI biosynthesis, resulting in MI production of ∼7.93 mg L–1. Finally, by optimizing the cultivation condition via supplying bicarbonate to enhance carbon fixation, a final MI production up to 12.72 mg L–1 was achieved, representing a ∼12-fold increase compared with the initial MI-producing strain. This study provides a light-driven green synthetic strategy for MI directly from CO2 in cyanobacterial chassis and represents a renewable alternative that may deserve further optimization in the future.

研究领域: 微生物学 · 生物技术 · 合成生物学
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BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS · 2023 · IF 3.1

Titanium dioxide nanoparticles enhance photocurrent generation of cyanobacteria

Yilan Li,Haowei Wang,Lingfang Tang,Huawei Zhu

Photosynthetic microorganisms such as cyanobacteria can convert photons into electrons, providing ideal eco-friendly materials for converting solar energy into electricity. However, the electrons are hardly transported outside the cyanobacterial cells due to the insulation feature of the cell wall/membrane. Various nanomaterials have been reported to enhance extracellular electron transfer of heterotrophic electroactive microorganisms, but its effect on intact photosynthetic microorganisms remains unclear. In this study, we investigated the effect of six different nanomaterials on the photocurrent generation of cyanobacterium Synechocystis sp. PCC 6803. Among the nanomaterials tested, titanium dioxide (TiO 2 ) nanoparticles increased the photocurrent generation of Synechocystis sp. PCC 6803 up to four-fold at the optimum concentration of 2 mg/mL. Transmission electron microscopy and scanning electron microscopy showed that TiO 2 bound to cyanobacterial cells and likely penetrated inside of cell membrane. Photochemical analyses for photosystems showed that TiO 2 blocked the electrons transfer downstream in PS I, implying a possible extracellular electron pathway mediated by TiO 2 . This study provides an alternative approach for enhancing the photocurrent generation of cyanobacteria, showing the potential of photosynthetic-nanomaterial hybrids.

研究领域: 生物技术 · 纳米技术 · 环境科学 · 微生物学
✓ 已验证 DOI PubMed 高影响因子
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Biotechnology for Biofuels · 2018 ·Vol. 11 ·期 1 ·1-16 · IF 5.497

Re-direction of carbon flux to key precursor malonyl-CoA via artificial small RNAs in photosynthetic Synechocystis sp. PCC 6803

Sun Tao,Li Shubin,Song Xinyu,Pei Guangsheng,Diao Jinjin,Cui Jinyu,Shi Mengliang,Chen Lei,Zhang Weiwen

Background Photosynthetic cyanobacteria have attracted a significant attention as promising chassis to produce renewable fuels and chemicals due to their capability to utilizing solar energy and CO 2 . Notably, the enhancing supply of key precursors like malonyl-CoA would benefit the production of many bio-compounds. Nevertheless, the lacking of genetic tools in cyanobacteria, especially the knockdown strategies for essential pathways, has seriously restricted the attempts to re-direct carbon flux from the central carbohydrate metabolism to the synthesis of bioproducts. Results Aiming at developing new genetic tools, two small RNA regulatory tools are reported for the model cyanobacterium Synechocystis sp. PCC6803, based on paired termini RNAs as well as the exogenous Hfq chaperone and MicC scaffold (Hfq-MicC) previously developed in Escherichia coli . Both regulatory tools functioned well in regulating exogenous reporter gene lacZ and endogenous glgC gene in Synechocystis sp. PCC6803, achieving a downregulation of gene expression up to 90% compared with wildtype. In addition, the Hfq-MicC tool was developed to simultaneously regulate multiple genes related to essential fatty acids biosynthesis, which led to decreased fatty acids content by 11%. Furthermore, aiming to re-direct the carbon flux, the Hfq-MicC tool was utilized to interfere the competing pathway of malonyl-CoA, achieving an increased intracellular malonyl-CoA abundance up to 41% (~ 698.3 pg/mL/OD 730 nm ) compared to the wildtype. Finally, the Hfq-MicC system was further modified into an inducible system based on the theophylline-inducible riboswitch. Conclusions In this study, two small RNA regulatory tools for manipulating essential metabolic pathways and re-directing carbon flux are reported for Synechocystis sp. PCC6803. The work introduces efficient and valuable metabolic regulatory strategies for photosynthetic cyanobacteria.

研究领域: 微生物学 · 代谢工程 · 合成生物学
✓ 已验证 DOI PubMed 高影响因子
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ACS Nano · 2024 · IF 15.8

Engineered Cyanobacteria-Based Living Materials for Bioremediation of Heavy Metals Both In Vitro and In Vivo

Tao Sun,Huaishu Huo,Yingying Zhang,Yaru Xie,Yize Li,Kungang Pan,Fenfang Zhang,Jing Liu,Yindong Tong,Weiwen Zhang,Lei Chen

The pollution caused by heavy metals (HMs) represents a global concern due to their serious environmental threat. Photosynthetic cyanobacteria have a natural niche and the ability to remediate HMs such as cadmium. However, their practical application is hindered by a low tolerance to HMs and issues related to recycling. In response to these challenges, this study focuses on the development and evaluation of engineered cyanobacteria-based living materials for HMs bioremediation. Genes encoding phytochelatins (PCSs) and metallothioneins (MTs) were introduced into the model cyanobacterium Synechocystis sp. PCC 6803, creating PM/6803. The strain exhibited improved tolerance to multiple HMs and effectively removed a combination of Cd2+, Zn2+, and Cu2+. Using Cd2+ as a representative, PM/6803 achieved a bioremediation rate of approximately 21 μg of Cd2+/OD750 under the given test conditions. To facilitate its controllable application, PM/6803 was encapsulated using sodium alginate-based hydrogels (PM/6803@SA) to create “living materials” with different shapes. This system was feasible, biocompatible, and effective for removing Cd2+ under simulated conditions of zebrafish and mice models. Briefly, in vitro application of PM/6803@SA efficiently rescued zebrafish from polluted water containing Cd2+, while in vivo use of PM/6803@SA significantly decreased the Cd2+ content in mice bodies and restored their active behavior. The study offers feasible strategies for HMs bioremediation using the interesting biomaterials of engineered cyanobacteria both in vitro and in vivo.

CAS / MDL: 7790-84-3 7601-90-3
研究领域: 环境科学 · 微生物学 · 生物技术 · 材料科学
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