Journal: International Journal of Molecular Sciences

Article Title: Semi-Biosynthetic Production of Surface-Binding Adhesive Antimicrobial Peptides Using Intein-Mediated Protein Ligation

doi: 10.3390/ijms232315202

Figure Lengend Snippet: Fed-batch fermentation of E. coli BL21 (DE3)-pET21b-HNG. ( a ) E. coli fed-batch fermentation kinetics for the production of HNG. Cells were cultivated in batch mode until the initially present glucose was depleted. After 5 h, glucose feeding was initiated at a predefined rate. At an OD 600 of 62, protein expression was induced with 0.1 mM IPTG. Samples were collected to monitor cell OD 600 (black) and medium glucose content (red). ( b ) Culture samples collected at induction times of 9, 12, 15, 18, 21, and 24 h were analyzed using SDS-PAGE and Western blotting with an anti-His antibody. Lane M: protein molecular weight marker; Lane W: whole cell lysate; Lane S: soluble fraction; Lane I: insoluble fraction. The target protein is indicated by arrows.

Article Snippet: E. coli strains DH5α and BL21 (DE3) (RBC Bioscience, Taipei, Taiwan) were used as hosts for cloning and HNG fusion protein expression, respectively. pET21b (Novagen, Madison, WI, USA) was used to construct expression plasmids.

Techniques: Expressing, SDS Page, Western Blot, Molecular Weight, Marker

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