Journal: BMC Biotechnology

Article Title: High yield production and purification of two recombinant thermostable phosphotriesterase-like lactonases from Sulfolobus acidocaldarius and Sulfolobus solfataricus useful as bioremediation tools and bioscavengers

doi: 10.1186/s12896-018-0427-0

Figure Lengend Snippet: Shake flask growth curves of E. coli-Sacpox ( a ) and E. coli-Ssopox 3 M ( b ) induced with 0.01, 0.1, 0.5, 1.0 mM IPTG, 5.0 mM or 10.0 mM galactose at about 1.0 Abs 600nm , as indicated by the arrows, compared with a not-induced growth; comparison of Sac Pox and Sso Pox 3 M enzyme production (U·g cww −1 ) in the different shake flask experiments ( c ). [*p < 0.05 compared to the not- induced shake flask; **p < 0.05 compared to the IPTG induced shake flask]

Article Snippet: The strains E. coli BL21 (DE3)- Sacpox and E. coli BL21 (DE3)- Ssopox 3 M (C258L/I261F/W263A) were obtained by Prof. Manco by employing genetic engineering strategies, as previously described [ , ], and they were stored at − 80 °C in 20% ( v /v) glycerol stock solutions.

Techniques: Comparison

Journal: BMC Biotechnology

Article Title: High yield production and purification of two recombinant thermostable phosphotriesterase-like lactonases from Sulfolobus acidocaldarius and Sulfolobus solfataricus useful as bioremediation tools and bioscavengers

doi: 10.1186/s12896-018-0427-0

Figure Lengend Snippet: Batch experiments (2.5 L) of E. coli-Sacpox and E. coli-Ssopox 3 M induced with 1.0 mM IPTG, 5.0 or 10.0 mM galactose at around 6.0 Abs 600nm , as indicated by the arrows: growth curves, IPTG or galactose up-take ( a - b ); Sac Pox and Sso Pox 3 M enzyme production (U·L − 1 ) in the different batch experiments ( c - d )

Article Snippet: The strains E. coli BL21 (DE3)- Sacpox and E. coli BL21 (DE3)- Ssopox 3 M (C258L/I261F/W263A) were obtained by Prof. Manco by employing genetic engineering strategies, as previously described [ , ], and they were stored at − 80 °C in 20% ( v /v) glycerol stock solutions.

Techniques:

Journal: BMC Biotechnology

Article Title: High yield production and purification of two recombinant thermostable phosphotriesterase-like lactonases from Sulfolobus acidocaldarius and Sulfolobus solfataricus useful as bioremediation tools and bioscavengers

doi: 10.1186/s12896-018-0427-0

Figure Lengend Snippet: Fed-batch experiments in 2.5 and 22.0-L vessels of E. coli Sacpox and E. coli Ssopox 3 M induced with 10.0 mM galactose at about 40.0 Abs 600nm , as indicated by the arrows: growth curves, galactose up-take, glycerol consumption, acetic acid formation and feeding profile ( a - b ). Sac Pox and Sso Pox 3 M enzyme production (U·L − 1 ) in the 2.5 and 22.0-L fed-batch experiments ( c - d )

Article Snippet: The strains E. coli BL21 (DE3)- Sacpox and E. coli BL21 (DE3)- Ssopox 3 M (C258L/I261F/W263A) were obtained by Prof. Manco by employing genetic engineering strategies, as previously described [ , ], and they were stored at − 80 °C in 20% ( v /v) glycerol stock solutions.

Techniques:

Journal: Journal of Advanced Research

Article Title: New insight into the mechanism by which antifreeze peptides regulate the physiological function of Streptococcus thermophilus subjected to freezing stress

doi: 10.1016/j.jare.2022.05.002

Figure Lengend Snippet: Expression and identification of recombinant snow flea antifreeze peptide (rsfAFP) in E. coli BL21 (DE3). (a) Target protein purification profile. Lanes 1–2: loaded sample; lane 3: flow through; lanes 4–6: fractions eluted with 20 mM TBS buffer with 20, 50, and 250 mM imidazole, respectively; lanes 7–8: 2 μg and 4 μg BSA (66.4 kDa), respectively; (b) Enzymatic hydrolysis of fusion proteins with SUMO protease. L1: SUMO; L2: SUMO-rsfAFP; L3: Marker; (c) Thermal hysteresis activity of rsfAFP at different concentrations; (d) Optical images show the completely different growth behaviors and shapes of ice crystals with or without the addition of 0.5 mg/mL rsfAFP in PBS (20 mM, pH 7.4). The scale is 100 μm; (e) Time-dependent NMR microimaging of frozen aqueous solutions during melting as labeled in the top left frame. Solutions containing rsfAFP at concentration of 0.5 mg/mL dissolved in 20 mM TBS (labeled rsfAFP1 in the figure), 5 mM TBS (labeled rsfAFP2) and pure water (labeled rsfAFP3) are shown. Pure water was used as the negative control, and 1.0 mg/mL arginine and 10% glycerol were used as positive controls. In the proton density images, black represents solid ice, and white represents areas with high densities of mobile water.

Article Snippet: The E. coli BL21 (DE3) expression strain was purchased from MoBiTec (Göettingen, Germany).

Techniques: Expressing, Recombinant, Protein Purification, Marker, Activity Assay, Labeling, Concentration Assay, Negative Control