Isolation and characterization of a thermophilic Bacillus shackletonii K5 from a biotrickling filter for the production of polyhydroxybutyrate

Polyhydroxyalkanoates（PHAs） are aliphatic polyesters accumulated intracellularly by both Gram-negative and Gram-positive bacteria. However, compared to the PHAs of Gramnegative bacteria, few endotoxins（lipopolysaccharides, LPS）, which would be co-purified with PHAs and cause immunogenic reactions, are found in the PHAs produced by Gram-positive bacteria. A thermophilic Gram-positive bacterium K5, which exhibited good growth and polyhydroxybutyrate（PHB）-accumulating ability, has been isolated and characterized from a biotrickling filter designed for the removal of NOx from flue gas in a coal-fired power plant in China. Based on the biochemical characterization and 16 S rRNA gene sequence（Genbank accession no. JX437933）, the strain K5 has been identified as Bacillus shackletonii, which has rarely been reported in the literature, and this report is the first time that B. shackletonii has been found to accumulate PHB. The strain K5 was able to utilize glucose as carbon source to synthesize PHB at a broad range of temperatures（from 35 to 50 °C）, and the ideal temperature was 45 °C. The strain K5 could effectively yield PHB of up to 69.9% of its cell dry weight（CDW）（2.28 g/L） in flask experiments employing glucose as carbon source at 45 °C, followed by 56.8% and 52.3% of its CDW when using sodium succinate and glycerol as carbon source, respectively. For batch cultivation, the strain K5 was able to produce PHB of up to 72.6% of its cell dry weight（9.76 g/L） employing glucose as carbon source at 45 °C and pH 7.0.     

Physiological cellular responses and adaptations of Rhodococcus erythropolis IBBPo1 to toxic organic solvents

A new Gram-positive bacterium, Rhodococcus erythropolis IBBPo1（KF059972.1） was isolated from a crude oil-contaminated soil sample by enrichment culture method. R. erythropolis IBBPo1 was able to tolerate a wide range of toxic compounds, such as antibiotics（800–1000 μg/mL）,synthetic surfactants（50–200 μg/mL）, and organic solvents（40%–100%）. R. erythropolis IBBPo1 showed good tolerance to both alkanes（cyclohexane, n-hexane, n-decane） and aromatics（toluene, styrene, ethylbenzene） with logPOW（logarithm of the partition coefficient of the solvent in octanol–water mixture） values between 2.64 and 5.98. However, alkanes were less toxic for R. erythropolis IBBPo1 cells, compared with aromatics. The high organic solvent tolerance of R. erythropolis IBBPo1 could be due to the presence in their large genome of some catabolic（alkB, alkB1, todC1, todM, xylM）, transporter（HAE1） and trehalose-6-phosphate synthase（otsA1, KF059973.1） genes. Numerous and complex physiological cellular responses and adaptations involved in organic solvent tolerance were revealed in R. erythropolis IBBPo1 cells exposed 1 and 24 hr to 1% organic solvents. R. erythropolis IBBPo1 cells adapt to 1% organic solvents by changing surface hydrophobicity, morphology and their metabolic fingerprinting.Considerable modifications in otsA1 gene sequence were also observed in cells exposed to organic solvents（except ethylbenzene）.