Document Detail


Highly efficient biotransformation of eugenol to ferulic acid and further conversion to vanillin in recombinant strains of Escherichia coli.
MedLine Citation:
PMID:  14602615     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The vaoA gene from Penicillium simplicissimum CBS 170.90, encoding vanillyl alcohol oxidase, which also catalyzes the conversion of eugenol to coniferyl alcohol, was expressed in Escherichia coli XL1-Blue under the control of the lac promoter, together with the genes calA and calB, encoding coniferyl alcohol dehydrogenase and coniferyl aldehyde dehydrogenase of Pseudomonas sp. strain HR199, respectively. Resting cells of the corresponding recombinant strain E. coli XL1-Blue(pSKvaomPcalAmcalB) converted eugenol to ferulic acid with a molar yield of 91% within 15 h on a 50-ml scale, reaching a ferulic acid concentration of 8.6 g liter(-1). This biotransformation was scaled up to a 30-liter fermentation volume. The maximum production rate for ferulic acid at that scale was 14.4 mmol per h per liter of culture. The maximum concentration of ferulic acid obtained was 14.7 g liter(-1) after a total fermentation time of 30 h, which corresponded to a molar yield of 93.3% with respect to the added amount of eugenol. In a two-step biotransformation, E. coli XL1-Blue(pSKvaomPcalAmcalB) was used to produce ferulic acid from eugenol and, subsequently, E. coli(pSKechE/Hfcs) was used to convert ferulic acid to vanillin (J. Overhage, H. Priefert, and A. Steinbüchel, Appl. Environ. Microbiol. 65:4837-4847, 1999). This process led to 0.3 g of vanillin liter(-1), besides 0.1 g of vanillyl alcohol and 4.6 g of ferulic acid liter(-1). The genes ehyAB, encoding eugenol hydroxylase of Pseudomonas sp. strain HR199, and azu, encoding the potential physiological electron acceptor of this enzyme, were shown to be unsuitable for establishing eugenol bioconversion in E. coli XL1-Blue.
Authors:
Jörg Overhage; Alexander Steinbüchel; Horst Priefert
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Publication Detail:
Type:  Evaluation Studies; Journal Article    
Journal Detail:
Title:  Applied and environmental microbiology     Volume:  69     ISSN:  0099-2240     ISO Abbreviation:  Appl. Environ. Microbiol.     Publication Date:  2003 Nov 
Date Detail:
Created Date:  2003-11-06     Completed Date:  2004-02-20     Revised Date:  2010-10-13    
Medline Journal Info:
Nlm Unique ID:  7605801     Medline TA:  Appl Environ Microbiol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  6569-76     Citation Subset:  IM    
Affiliation:
Institut für Molekulare Mikrobiologie und Biotechnologie der Westfälischen Wilhelms-Universität Münster, D-48149 Münster, Germany.
Data Bank Information
Bank Name/Acc. No.:
GENBANK/AJ006231;  Y15627
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MeSH Terms
Descriptor/Qualifier:
Alcohol Oxidoreductases / chemistry,  genetics,  metabolism
Aldehyde Oxidoreductases / chemistry,  genetics,  metabolism
Bacterial Proteins*
Benzaldehydes / metabolism
Coumaric Acids / metabolism*
Escherichia coli / genetics,  metabolism*
Eugenol / metabolism*
Molecular Sequence Data
Penicillium / enzymology,  genetics
Recombinant Proteins / genetics,  metabolism
Sequence Analysis, DNA
Chemical
Reg. No./Substance:
0/Bacterial Proteins; 0/Benzaldehydes; 0/Coumaric Acids; 0/Recombinant Proteins; 1135-24-6/ferulic acid; 121-33-5/vanillin; 97-53-0/Eugenol; EC 1.1.-/Alcohol Oxidoreductases; EC 1.1.1.-/coniferyl alcohol dehydrogenase; EC 1.1.3.38/vanillyl-alcohol oxidase; EC 1.2.-/Aldehyde Oxidoreductases; EC 1.2.1.-/coniferyl aldehyde dehydrogenase
Comments/Corrections

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