Gene cloning of phenolic acid decarboxylase from Bacillus subtilis and expression in top-fermenting yeast strain

  • Xiaohong Cao
  • Yunqian Cui
  • Xinghua Liao
  • Guangtian Zhou
  • Leo Thamm

Abstract

Phenolic acid decarboxylase (PADC) gene, encoding phenolic acid decarboxylase, was cloned from Bacillus subtilis and ligated with a shuttle vector YEp352 to generate a novel plasmid YPADC. By analysis of sequencing and the restriction endonuclease digestion, the validity of construction was proved. Subsequently, the new vector was successfully transformed into wild-type top-fermenting yeast strain W303-1A; the mutant yeast strain W303+padc was obtained, which was tested on the laboratoryscale mashing and fermentation experiments. At the end of fermentation, the results showed an obvious increase of 4-vinylguaiacol content in top-fermented beers brewed with mutant yeasts. The final 4-vinylguaiacol concentration obtained with wild-type and mutant yeasts was 1.20 and 1.70mg/l, respectively. Additionally, the level of esters produced by the mutant strain was higher than that of the wild-type; there were therefore a marked clove-like and ester aroma in top-fermented beers brewed with the former. However, no evident differences were found in brewing characteristic between wild-type and mutant strains, especially the ability of utilizing fermentable sugar and reducing diacetyl. Taken together, these approaches indicated the possibility of cloning PADC gene and enhancing the concentration of 4-vinylguaiacol in top-fermented beers.size.

Keywords: Clone, phenolic acid decarboxylase, top-fermenting yeast, 4-vinylguaiacol

African Journal of Biotechnology Vol. 9(33), pp. 5284-5291, 16 August, 2010

Author Biographies

Xiaohong Cao
College of Food Engineering and Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, P. R. China
Yunqian Cui
College of Food Engineering and Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, P. R. China; China-Germany Brewing Technical Service Center, Shandong Institute of Light Industry, Jinan 250353, P. R. China
Xinghua Liao
College of Food Engineering and Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, P. R. China
Guangtian Zhou
China-Germany Brewing Technical Service Center, Shandong Institute of Light Industry, Jinan 250353, P. R. China
Leo Thamm
China-Germany Brewing Technical Service Center, Shandong Institute of Light Industry, Jinan 250353, P. R. China
Published
2013-08-09
Section
Articles

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eISSN: 1684-5315