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Fig. 1 | Microbial Cell Factories

Fig. 1

From: Large-scale production of a thermostable Rhodothermus marinus cellulase by heterologous secretion from Streptomyces lividans

Fig. 1

Heterologous synthesis of CelA in E. coli and synthesis/secretion in S. lividans. a CelAHis6 was synthesized in BL21 (DE3) cells. Expression of CelAHis6 carried on a pET vector was induced by 0.2 mM IPTG for 3 h at 30 °C. b Synthesis in and secretion of CelA from S. lividans. Polypeptides (1–8 µg/lane) from culture supernatants (~ 0.1 mL) that is equivalent to 0.2 mg of dry cell mass. The samples from cells grown for 2 days in the indicated media (see “Methods”) were harvested by precipitation with TCA (25%), analyzed by SDS-PAGE and stained with Coomassie blue. Lane 1, molecular weight markers: β-galactosidase (116 kDa), bovine serum albumin (66.2 kDa), ovalbumin (45 kDa), lactate dehydrogenase (35 kDa), restriction endonuclease Bsp98I (25 kDa), β-lactoglobulin (18.4 kDa), lysozyme (14.4 kDa). Pha, phage medium; NB, nutrient broth; NB2X, nutrient broth double strength; MM, minimal medium; C5 and C15, casamino acids (5 or 15 g/L); TSB, tryptic soy broth; Ben, Bennet medium

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