Parallel substrate supply and pH stabilization for optimal screening of E. coli with the membrane-based fed-batch shake flask

Abstract Background: R-(-)-linalool is a versatile acyclic monoterpene alcohol with applications in the flavor and fragrance, pharmaceutical, and agrochemical industries. However, plant extraction furnishes only limited and unstable R-(-)-linalool yields that do not satisfy market demand. Therefore, a sustainable yet efficient and productive method of R-(-)-linalool synthesis is urgently needed. Results: To induce the R-(-)-linalool biosynthesis pathway in E. coli , we expressed heterologous (3R)-linalool synthase (LIS) from Lavandula angustifolia (laLIS). We then enhanced R-(-)-linalool production in the cells using a suitable LIS from Streptomyces clavuligerus (bLIS). The bLIS expression was markedly elevated by using optimized ribosomal binding sites (RBSs) and protein fusion tags. R-(-)-linalool output rose from 4.8 mg L -1 to 33.4 mg L -1 . To increase the geranyl diphosphate (GPP) content in E. coli , we tested various alterations in geranyl diphosphate synthases (GPPSs) and their mutants. The final E. coli strain harboring GPPS from Abies grandis ( Ag GPPS) accumulated ≤ 100.1 mg L -1 R-(-)-linalool after 72 h shake-flask fermentation. This yield gain constitutes a 60.7-fold improvement in R-(-)-linalool biosynthesis over the parent strain. Fed-batch cultivation of the engineered strain in a 1.3-L fermenter yielded 1,027.3 mg L -1 R-(-)-linalool. Conclusions: In this study, an efficient R-(-)-linalool production pathway was induced in E. coli via the heterologous MVA pathway, AgGPPS, and (3R)-linalool synthase (bLIS). By overexpressing the key enzyme in the engineered E. coli strain, R-(-)-linalool production reached 100.1 mg L -1 and 1,027.3 mg L -1 under shake flask- and fed-batch fermentation conditions, respectively. The latter is the highest reported R-(-)-linalool yield to date using an engineered E. coli strain. The strategies of key enzyme overexpression and mutation could lay theoretical and empirical foundations for engineering terpenoid pathways and optimizing other metabolic pathways.

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A Novel Method for Preparation of Antimicrobial Cellulose Textile by Surface Carbamatization and N-Chlorination

Advanced Materials Research ◽

10.4028/www.scientific.net/amr.399-401.1088 ◽

2011 ◽

Vol 399-401 ◽

pp. 1088-1092

Author(s):

Liu Yang Wang ◽

Fang Ying Wang ◽

Biao Bing Wang

Keyword(s):

Nitrogen Content ◽

Shake Flask ◽

Antimicrobial Activities ◽

Sodium Hypochlorite Solution ◽

E Coli ◽

Cellulose Fabric ◽

Carbamate Group ◽

Novel Method ◽

Cellulose Fabrics ◽

Temperature Time

Surface carbamatization of cellulose fabric was accomplished through co-heating with urea. The carbamate group on textile surface was transformed to N-chlorocarbamate after exposure to sodium hypochlorite solution. Effect of carbamatization temperature, time on Nitrogen content of fabrics was investigated, and the effect of bleach technology on chlorine content on fabrics was also studied. Using shake flask method antimicrobial activities against E. coli and S. aureus were studied. The results showed that when carbamatization temperature was 138°C and the time was 3 hours, nitrogen content of cellulose fabric was the highest: 1.47%. Antibacterial ability of cellulose fabrics with 0.81mg/g chlorine against E. coli and S. aureus was 99.9999% and 100%, respectively.

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DESIGN OF ON-LINE STATE ESTIMATORS FOR A RECOMBINANT E. COLI FED-BATCH FERMENTATION

IFAC Proceedings Volumes ◽

10.3182/20050703-6-cz-1902.02215 ◽

2005 ◽

Vol 38 (1) ◽

pp. 67-72 ◽

Cited By ~ 2

Author(s):

Ana C.A. Veloso ◽

I. Rocha ◽

E.C. Ferreira

Keyword(s):

Batch Fermentation ◽

Fed Batch ◽

E Coli ◽

State Estimators ◽

Fed Batch Fermentation ◽

On Line

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Different induction regimes in recombinant E. coli fed batch cultivations to modulate inclusion body formation, disassembly or prevention

New Biotechnology ◽

10.1016/j.nbt.2009.06.165 ◽

2009 ◽

Vol 25 ◽

pp. S213

Author(s):

M. Cserjan-Puschmann ◽

M. Luchner ◽

F. Clementschitsch ◽

G. Striedner ◽

K. Bayer

Keyword(s):

Inclusion Body ◽

Fed Batch ◽

Body Formation ◽

E Coli ◽

Inclusion Body Formation

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Differential expression of small RNAs under chemical stress and fed-batch fermentation in E. coli

BMC Genomics ◽

10.1186/s12864-015-2231-8 ◽

2015 ◽

Vol 16 (1) ◽

Cited By ~ 14

Author(s):

Martin Holm Rau ◽

Klara Bojanovič ◽

Alex Toftgaard Nielsen ◽

Katherine S. Long

Keyword(s):

Differential Expression ◽

Small Rnas ◽

Batch Fermentation ◽

Chemical Stress ◽

Fed Batch ◽

E Coli ◽

Fed Batch Fermentation

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Validation of the transferability of membrane-based fed-batch shake flask cultivations to stirred-tank reactor using three different protease producing Bacillus strains

Journal of Bioscience and Bioengineering ◽

10.1016/j.jbiosc.2019.05.003 ◽

2019 ◽

Vol 128 (5) ◽

pp. 599-605 ◽

Cited By ~ 2

Author(s):

Janina Müller ◽

Anne Hütterott ◽

Tobias Habicher ◽

Nina Mußmann ◽

Jochen Büchs

Keyword(s):

Shake Flask ◽

Stirred Tank ◽

Fed Batch ◽

Stirred Tank Reactor ◽

Tank Reactor ◽

Bacillus Strains

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The Rocky Road From Fed-Batch to Continuous Processing With E. coli

Frontiers in Bioengineering and Biotechnology ◽

10.3389/fbioe.2019.00328 ◽

2019 ◽

Vol 7 ◽

Cited By ~ 6

Author(s):

Julian Kopp ◽

Christoph Slouka ◽

Oliver Spadiut ◽

Christoph Herwig

Keyword(s):

Continuous Processing ◽

Fed Batch ◽

E Coli

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Co-Expression of Threonine Dehydratase and Acetolactate Synthase in Escherichia Coli for L-Isoleucine Production

Advanced Materials Research ◽

10.4028/www.scientific.net/amr.989-994.997 ◽

2014 ◽

Vol 989-994 ◽

pp. 997-1002 ◽

Cited By ~ 1

Author(s):

Jian Wang ◽

Jia Kai Sun ◽

Qing Yang Xu

Keyword(s):

Escherichia Coli ◽

Corynebacterium Glutamicum ◽

Carbon Flux ◽

Batch Fermentation ◽

Acetolactate Synthase ◽

Fed Batch ◽

Threonine Dehydratase ◽

E Coli ◽

Fed Batch Fermentation ◽

Fermentation Period

Metabolic engineering ofCorynebacterium glutamicumhas sought to divert carbon into L-isoleucine. However, the fermentation period of this strain is long. TheC.glutamicumYILW strain (LeuL, AHVr, SGr, Leu-MEr) was previously derived by repeated compound mutagenesis which could accumulate 20.2 g/L L-isoleucine in a 5-L jar fermentor. Overexpression of the threonine dehydratase gene (ilvA) fromCorynebacterium glutamicumYILW and coexpression of threonine dehydratase and acetolactate synthase (ilvBN) from it were employed to divert carbon flux toward L-isoleucine. The strainE. coliTRFC with the expression ofilvA could accumulate L-isoleucine of 6.8 g/L without accumulation of any L-threonine by fed-batch fermentation in a 5-L jar fermentor. However, the production of L-isoleucine by the strainE.coliTRFC with the co-expression ofilvA andilvBN was decreased by 19.1%, and the production of L-valine was increased by 40% compared with that ofE. coliTRFC with the expression ofilvA.

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