Production of cyclic adenosine-3′,5′-monophosphate by whole cell catalysis using recombinant Escherichia coli overexpressing adenylate cyclase

Li, Nan; Ying, Hanjie; Chen, Yong; Chen, Xiaochun; Bai, Jianxin; Wu, Jinglan; Xie, Jingjing; Ying, Hanjie

doi:10.1007/s11814-012-0202-1

Cited by 5 publications

(3 citation statements)

References 27 publications

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“…Researches in recent years were conducted and demonstrated that many microorganisms could be used to produce cAMP. Report found that 14.93 g/L of cAMP was produced with a conversion rate of 91.5% when pyruvate sodium was used to be enzyme activator for 6 hr in Escherichia coli (Li et al, ). It also found that the addition of 0.4 g/L of sodium fluoride in Arthrobacter A302, the maximal cAMP concentration reached 11.04 g/L (Chen et al, ).…”

Section: Introductionmentioning

confidence: 99%

Construction of industrial baker’s yeast with high level of cAMP

Dong

Wang

et al. 2019

J Food Biochem

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Cyclic adenosine monophosphate (cAMP) plays an important role in modulating the activity of microbe cell. In this study, PKA (protein kinase A) activity was weakened through truncation of TPK2 promoter (−150 bp and −300 bp) and gene deletion of BCY1 (encodes the regulatory subunit of PKA), TPK1 and TPK3, generating strains BY9a‐T2‐150 and BY9a‐T2‐300, respectively. High‐performance liquid chromatography analysis showed cAMP levels in BY9a‐T2‐150 and BY9a‐T2‐300 were increased by 5‐ and 18‐fold, respectively, compared with that of parent strain, BY9a. The expression levels of TPK2 gene in two engineered strains were decreased by 95% and 97% compared with that of BY9a, respectively. The PKA activity reflected by heat resistance of engineered strains enhanced compared with parent strain BY9a. This study show a new method to increase the intracellular cAMP concentration in industrial yeast by fine‐tuning of PKA activity, without influence in growth and fermentation properties. Practical applications cAMP as the “second messenger,” is essential for plant, animal, and microorganisms and human life. But its synthesis is still limited by expensive cost and time‐consuming method. We constructed the industrial baker’s yeast with high level of cAMP and desired to be used to produce functional food for relaxing smooth muscle, expanding blood vessels, improving liver function, and promoting nerve regeneration and as a food additive for treating hyperthyreosis and hepatopathy. The methods of two step homologous recombination and backcross operated in this study eliminate the exogenous gene in engineered strains, made it safety to be used in food production. Fine‐tuning of PKA activity in engineered strains ensure produce high level of cAMP and exhibit normal growth performance in engineering strains. Therefore, this work is significant in functional foods product and has the potential to be used in practical application.

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Section: Introductionmentioning

confidence: 99%

Construction of industrial baker’s yeast with high level of cAMP

Dong

Wang

et al. 2019

J Food Biochem

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“…Because biotechnological applications of OPH require economical production of the enzyme with high activity, there have been many efforts to produce OPH in a soluble, active form using bacterial expression systems. A practical approach might be to make bacterial whole‐cell biocatalytic system for more economical use of the enzymes . Whole‐cell system holds great promise as a potential biocatalyst owing to several advantages such as relatively high stability, easy purification, low preparation cost, and efficient cofactor regeneration compared with free enzyme system .…”

Section: Introductionmentioning

confidence: 99%

“…A practical approach might be to make bacterial whole-cell biocatalytic system for more economical use of the enzymes. [6][7][8][9][10][11][12][13][14] Wholecell system holds great promise as a potential biocatalyst owing to several advantages such as relatively high stability, †Dong Gyun Kang and Jeong Hyun Seo contributed equally to this work.…”

Section: Introductionmentioning

confidence: 99%

Versatile signal peptide of Flavobacterium‐originated organophosphorus hydrolase for efficient periplasmic translocation of heterologous proteins in Escherichia coli

Kang

Seo

et al. 2016

Biotechnology Progress

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Organophosphorus hydrolase (OPH) from Flavobacterium species is a membrane-associated homodimeric metalloenzyme and has its own signal peptide in its N-terminus. We found that OPH was translocated into the periplasmic space when the original signal peptide-containing OPH was expressed in recombinant Escherichia coli even though its translocation efficiency was relatively low. To investigate the usability of this OPH signal peptide for periplasmic expression of heterologous proteins in an E. coli system, we employed green fluorescent protein (GFP) as a cytoplasmic folding reporter and alkaline phosphatase (ALP) as a periplasmic folding reporter. We found that the OPH signal peptide was able to use both twin-arginine translocation (Tat) and general secretory (Sec) machineries by switching translocation pathways according to the nature of target proteins in E. coli. These results might be due to the lack of Sec-avoidance sequence in the c-region and a moderate hydrophobicity of the OPH signal peptide. Interestingly, the OPH signal peptide considerably enhanced the translocation efficiencies for both GFP and ALP compared with commonly used TorA and PelB signal peptides that have Tat and Sec pathway dependences, respectively. Therefore, this OPH signal peptide could be successfully used in recombinant E. coli system for efficient periplasmic production of target protein regardless of the subcellular localization where functional folding of the protein occurs. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:848-854, 2016.

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Efficient production of cyclic adenosine monophosphate from adenosine triphosphate by the N-terminal half of adenylate cyclase from Escherichia coli

Wang

et al. 2020

Chinese Journal of Chemical Engineering

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Production of cyclic adenosine-3′,5′-monophosphate by whole cell catalysis using recombinant Escherichia coli overexpressing adenylate cyclase

Cited by 5 publications

References 27 publications

Construction of industrial baker’s yeast with high level of cAMP

Construction of industrial baker’s yeast with high level of cAMP

Versatile signal peptide of Flavobacterium‐originated organophosphorus hydrolase for efficient periplasmic translocation of heterologous proteins in Escherichia coli

Efficient production of cyclic adenosine monophosphate from adenosine triphosphate by the N-terminal half of adenylate cyclase from Escherichia coli

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