European Food Research and Technology

, Volume 235, Issue 5, pp 951–961 | Cite as

Construction of recombinant industrial brewer’s yeast with lower diacetyl production and proteinase A activity

  • Jun Lu
  • Jian Dong
  • Deguang Wu
  • Yefu Chen
  • Xuewu Guo
  • Yu Shi
  • Xi Sun
  • Dongguang Xiao
Original Paper


The characteristic buttery taste of diacetyl has long been a major problem in the brewing industry, and the foam stability of unpasteurized beer is often influenced by proteinase A (PrA), which is encoded by PEP4 and released from yeast cells into beer during brewing. A recombinant industrial brewer’s yeast strain that reduces the diacetyl content of beer and improves foam stability was constructed. We constructed a PGK1p-ILV5-PGK1t expression cassette, which was introduced into one of the PEP4 alleles via PCR-mediated homologous recombination. Then, the second PEP4 allele was disrupted using the Cre-loxP recombination system, and the recombinant strain was designated as S-CSIK12. The results show that the diacetyl production of S-CSIK12 is always lower than that of the host strain at all stages of beer fermentation. In addition, brewing with S-CSIK12 reduced the PrA activity of the final beer by 44 % compared with that using the wild-type strain. The head retention of the beer brewed with S-CSIK12 (260 ± 2 s) was better than that of the host strain S-6 (212 ± 3 s). Considering that more PrA is released from yeast cells during the final stage of main fermentation and that the timing of yeast cropping is determined by diacetyl reduction, brewing with strains that have low diacetyl production also reduced the PrA activity of the beer and improved its head retention. The present study provides reference for the brewing industry as well as research on the diacetyl reduction and foam stability of beer.


Proteinase A (PrA) Diacetyl Head retention Industrial brewer’s yeast 



Proteinase A


A kind of gene that encodes proteinase A


A kind of gene that encodes reductoisomerase


Acetohydroxyacid reductoisomerase

ILV pathway

Isoleucine–valine pathway


α-Acetolactate decarboxylase


A kind of gene that encodes acetolactate synthase I/II/III large subunit


A kind of gene that encodes dihydroxy-acid dehydratase


A kind of gene that encodes acetolactate synthase I/III small subunit


Lipid transfer protein 1


A kind of gene that encodes glycerol-3-phosphate dehydrogenase


Reference dye for real-time PCR






Pyruvate kinase


Wild-type industrial brewer’s yeast


Two PEP4 allele disruption strain


Two PEP4 allele disruption and ILV5 overexpression strain



The current study was financially supported by the program for Changjiang Scholars and Innovative Research Team in University (IRT1166), the National Natural Science Foundation of China (No.31271916).


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Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Jun Lu
    • 1
  • Jian Dong
    • 1
  • Deguang Wu
    • 1
  • Yefu Chen
    • 1
  • Xuewu Guo
    • 1
  • Yu Shi
    • 1
  • Xi Sun
    • 1
  • Dongguang Xiao
    • 1
  1. 1.Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin Industrial Microbiology Key Laboratory, College of BiotechnologyTianjin University of Science and TechnologyTianjinPeople’s Republic of China

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