• Microbiology and Biotechnology Letters
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• v.44 no.3
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• pp.341-348
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• 2016

This study aimed to develop a microbial process for producing aminolevulinic acid (ALA) using crude glycerol. In the culture of ALA-producing cells (Escherichia coli/pH-hemA) in a medium containing crude glycerol, the cell density and production were 1.8-fold and 1.2-fold lower than those obtained from pure glycerol, respectively. However, the cell growth and production were improved by supplementing the medium with trehalose (30 or 100 g/l). Engineered cells (E. coli/pH-hemA/pS-otsBA) were constructed to express otsBA and their culture performance was compared with that of control cells (E. coli/pH-hemA/ pSTV28). The effects of isopropyl β-D-1-thiogalactopyranoside (IPTG) concentration and the time of induction were examined to improve the cell growth and ALA production in engineered cells cultured using crude glycerol. When 0.6 mM of IPTG was added at the beginning of the exponential growth phase, the ALA produced by cells was 2,121 mg/l, which was comparable to that from pure glycerol. The results demonstrate that otsBA expression endowed cells with the capacity to tolerate the toxicity of crude glycerol for direct use.

• Microbiology and Biotechnology Letters
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• v.44 no.4
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• pp.497-503
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• 2016

This study aimed to optimize the culture conditions of recombinant Escherichia coli expressing otsBA using crude glycerol for the enhanced production of trehalose. The effects of culture temperature and isopropyl ${\beta}$-D-1-thiogalactopyranoside (IPTG)-induction were investigated. Trehalose production and cell growth were highest when cells were cultured at $37^{\circ}C$ and induced with IPTG. The concentrations of IPTG, validamycin A, and NaCl were optimized using Box-Behnken design. Statistical analyses of the experimental data revealed that the concentrations of IPTG and NaCl had significant effects on trehalose production, but that of validamycin A did not. Contour plot analysis and model calculation showed that the highest amount of trehalose could be produced at 298 mM NaCl and 0.1 mM IPTG. Under these optimal conditions, the optical density at 600 nm and trehalose production were $5.4{\pm}0.2$ and $304{\pm}15mg/l$, respectively.

• Journal of Microbiology and Biotechnology
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• v.23 no.11
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• pp.1560-1568
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• 2013

Salmonella, a main cause of foodborne diseases, encounters a variety of environmental stresses and overcomes the stresses by multiple resistance strategies. One of the general responses to hyperosmotic stress is to import or produce compatible solutes so that cells maintain fluid balance and protect proteins and lipids from denaturation. The ProP and ProU systems are the main transport systems for compatible solutes. The OsmU system, recently identified as a third osmoprotectant transport system, debilitates excessive growth as well by reducing production of trehalose. We studied a fourth putative osmoprotectant transport system, YehZYXW, with high sequence similarity with the OsmU system. A Salmonella strain lacking YehZ, a predicted substrate-binding protein, did not suffer from hyperosmolarity but rather grew more rapidly than the wild type regardless of glycine betaine, an osmoprotectant, suggesting that the YehZYXW system controls bacterial growth irrespective of transporting glycine betaine. However, the growth advantage of ${\Delta}yehZ$ was not attributable to an increase in OtsBA-mediated trehalose production, which is responsible for the outcompetition of the ${\Delta}osmU$ strain. Overexpressed YehZ in trans was capable of deaccelerating bacterial growth vice versa, supporting a role of YehZ in dampening growth. The expression of yehZ was increased in response to nutrient starvation, acidic pH, and the presence of glycine betaine under hyperosmotic stress. Identifying substrates for YehZ will help decipher the role of the YehZYXW system in regulating bacterial growth in response to environmental cues.