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  • Metabolomic basis of laboratory evolution of butanol tolerance in photosynthetic Synechocystis sp. PCC 6803.

Metabolomic basis of laboratory evolution of butanol tolerance in photosynthetic Synechocystis sp. PCC 6803.

Microbial cell factories (2014-11-05)
Yaxing Wang, Mengliang Shi, Xiangfeng Niu, Xiaoqing Zhang, Lianju Gao, Lei Chen, Jiangxin Wang, Weiwen Zhang
ABSTRACT

Recent efforts demonstrated the potential application of cyanobacteria as a "microbial cell factory" to produce butanol directly from CO2. However, cyanobacteria have very low tolerance to the toxic butanol, which limits the economic viability of this renewable system. Through a long-term experimental evolution process, we achieved a 150% increase of the butanol tolerance in a model cyanobacterium Synechocystis sp. PCC 6803 after a continuous 94 passages for 395 days in BG11 media amended with gradually increased butanol concentration from 0.2% to 0.5% (v/v). To decipher the molecular mechanism responsible for the tolerance increase, we employed an integrated GC-MS and LC-MS approach to determine metabolomic profiles of the butanol-tolerant Synechocystis strains isolated from several stages of the evolution, and then applied PCA and WGCNA network analyses to identify the key metabolites and metabolic modules related to the increased tolerance. The results showed that unstable metabolites of 3-phosphoglyceric acid (3PG), D-fructose 6-phosphate (F6P), D-glucose 6-phosphate (G6P), NADPH, phosphoenolpyruvic acid (PEP), D-ribose 5-phosphate (R5P), and stable metabolites of glycerol, L-serine and stearic acid were differentially regulated during the evolution process, which could be related to tolerance increase to butanol in Synechocystis. The study provided the first time-series description of the metabolomic changes related to the gradual increase of butanol tolerance, and revealed a metabolomic basis important for rational tolerance engineering in Synechocystis.

MATERIALS
Product Number
Brand
Product Description

Supelco
1-Butanol, analytical standard
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1-Butanol, for molecular biology, ≥99%
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Supelco
1-Butanol, Pharmaceutical Secondary Standard; Certified Reference Material
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1-Butanol, 99.9%
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1-Butanol, ACS reagent, ≥99.4%
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1-Butanol, suitable for HPLC, ≥99.7%
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Butyl alcohol, natural, ≥99.5%, FCC, FG