Journal: Microbial Cell Factories

Article Title: Improved productivity of poly (4-hydroxybutyrate) (P4HB) in recombinant Escherichia coli using glycerol as the growth substrate with fed-batch culture

doi: 10.1186/s12934-014-0131-2

Figure Lengend Snippet: Influence of various factors on growth and P4HB accumulation. E. coli JM109 (pKSSE5.3) was grown in 1 L bioreactors in modified E2 medium. The modified E2 medium containing 10 g L −1 xylose, 4 g L −1 Na-4HB, 1 g L −1 NZ-amines, and 1 mL L −1 trace elements was used as standard medium. Culture A : standard medium; Culture B : two times more xylose was added to the standard medium, leading to 20 g L −1 xylose; Culture C : five times more nitrogen source NaNH 4 HPO 4 · 4H 2 O was added to the standard medium, leading to a final NaNH 4 HPO 4 · 4H 2 O concentration of 17.5 g L −1 ; Culture D : NZ-amine amount was increased by 5 fold, leading to 5 g L −1 ; Culture E : three times more trace elements were added, leading to 3 mL L −1 ; Culture F : three times more magnesium was added, leading to 3 mM of MgSO 4 · 7H 2 O. The data are the average numbers of duplicates.

Article Snippet: E. coli JM109 (pKSSE5.3) cells were grown at 32°C in 1 L bioreactors (Infors AG, Bottmingen, Switzerland) containing modified E2 medium supplemented with 10 g L −1 xylose, 4 g L −1 Na-4HB, 1 g L −1 NZ-amines and 0.015 g L −1 thiamine.

Techniques: Modification, Concentration Assay

Journal: Microbial Cell Factories

Article Title: Improved productivity of poly (4-hydroxybutyrate) (P4HB) in recombinant Escherichia coli using glycerol as the growth substrate with fed-batch culture

doi: 10.1186/s12934-014-0131-2

Figure Lengend Snippet: E . coli JM109 (pKSSE5.3) grown in modified E2 medium supplemented with or without acetate in shake flasks. 10 g L −1 glycerol was used as the main carbon source. A : 2 g L −1 acetate was added to the culture; B : 1 g L −1 acetate was added to the culture. The data are the average numbers of duplicates.

Article Snippet: E. coli JM109 (pKSSE5.3) cells were grown at 32°C in 1 L bioreactors (Infors AG, Bottmingen, Switzerland) containing modified E2 medium supplemented with 10 g L −1 xylose, 4 g L −1 Na-4HB, 1 g L −1 NZ-amines and 0.015 g L −1 thiamine.

Techniques: Modification

Journal: Microbial Cell Factories

Article Title: Improved productivity of poly (4-hydroxybutyrate) (P4HB) in recombinant Escherichia coli using glycerol as the growth substrate with fed-batch culture

doi: 10.1186/s12934-014-0131-2

Figure Lengend Snippet: Fed-batch strategy using acetic acid as stimulator for P4HB synthesis in E . coli JM109 (pKSSE5.3) grown on modified M9 medium supplemented with 20 g L −1 glycerol, 6 g L −1 Na-4HB, 0.5 g L −1 NZ-amine, 0.015 g L −1 thiamine and 100 mg L −1 ampicillin. Acetic acid was added at different physiological states. For all cultures pulse-feeding started at 40 h of cultivation: T = 40.5 h, addition of 12 g L −1 glycerol and 6 g L −1 Na-4HB; T = 45.75 h, addition of 20 g L −1 glycerol; T = 63.75 h, addition of 10 g L −1 glycerol and 3 g L −1 Na-4HB; T = 72.25 h, addition of 20 g L −1 glycerol and 6 g L −1 Na-4HB; T = 76.75 h, addition of 10 g L −1 glycerol. Culture A , addition of 2 g L −1 acetic acid at the beginning of the cultivation; Culture B , addition of 1 g L −1 acetic acid at beginning and at the end of growth phase (66 h), respectively; Culture C , addition of 2 g L −1 acetic acid after 48 h cultivation; Culture D , no addition of acetic acid to the culture. Arrows represent the addition of acetic acid. The data are the average numbers of duplicates.

Article Snippet: E. coli JM109 (pKSSE5.3) cells were grown at 32°C in 1 L bioreactors (Infors AG, Bottmingen, Switzerland) containing modified E2 medium supplemented with 10 g L −1 xylose, 4 g L −1 Na-4HB, 1 g L −1 NZ-amines and 0.015 g L −1 thiamine.

Techniques: Modification

Journal: Microbial Cell Factories

Article Title: Improved productivity of poly (4-hydroxybutyrate) (P4HB) in recombinant Escherichia coli using glycerol as the growth substrate with fed-batch culture

doi: 10.1186/s12934-014-0131-2

Figure Lengend Snippet: Time course of cell dry weight (CDW), P4HB content and P4HB concentration during a pulse feeding fed-batch culture of E . coli JM109 (pKSSE5.3). Glycerol and 4-hydroxybutyric acid were used as carbon source and precursor, respectively. Cultivation was conducted at 32°C in a 1 L bioreactor with an initial volume of 600 mL of modified M9 medium plus 20 g L −1 glycerol, 6 g L −1 Na-4HB, 2 g L −1 acetic acid, 0.5 g L −1 NZ-amines, 0.015 g L −1 thiamine and 100 mg L −1 ampicillin. Feeding solution was added to the bioreactor when glycerol in the medium was close to depletion, indicated by pO 2 signal. Arrow #1, feeding of 12.5 g L −1 glycerol + 2.5 g L −1 Na-4HB + 1.1 g L −1 acetate at 30 h; Arrow #2, feeding of 25.0 g L −1 glycerol + 2.1 g L −1 acetate at 39 h; Arrow #3, feeding of 2.5 g L −1 Na-4HB at 41 h; Arrow #4, feeding of 25.0 g L −1 glycerol + 2.1 g L −1 acetate + 2.5 g L −1 Na-4HB at 44 h; Arrow #5, feeding of 25.0 g L −1 glycerol + 2.1 g L −1 acetate + 6 g L −1 Na-4HB at 49.5 h. The data are the average numbers of duplicates.

Article Snippet: E. coli JM109 (pKSSE5.3) cells were grown at 32°C in 1 L bioreactors (Infors AG, Bottmingen, Switzerland) containing modified E2 medium supplemented with 10 g L −1 xylose, 4 g L −1 Na-4HB, 1 g L −1 NZ-amines and 0.015 g L −1 thiamine.

Techniques: Concentration Assay, Modification

Journal: Microbial Cell Factories

Article Title: Improved productivity of poly (4-hydroxybutyrate) (P4HB) in recombinant Escherichia coli using glycerol as the growth substrate with fed-batch culture

doi: 10.1186/s12934-014-0131-2

Figure Lengend Snippet: High cell density cultivation with linear feeding mode of E . coli JM109 (pKSSE5.3). The feeding solution contained 200 g L −1 acetic acid and 200 g L −1 glycerol for 65 h and then 100 g L −1 acetic acid and 200 g L −1 glycerol. Panel A : feeding rate of 1 mL h −1 ; B : feeding rate of 2 mL h −1 . Arrows represent the start of feeding. The data are the average numbers of duplicates.

Article Snippet: E. coli JM109 (pKSSE5.3) cells were grown at 32°C in 1 L bioreactors (Infors AG, Bottmingen, Switzerland) containing modified E2 medium supplemented with 10 g L −1 xylose, 4 g L −1 Na-4HB, 1 g L −1 NZ-amines and 0.015 g L −1 thiamine.

Techniques: