Background: Enhancing the industrial yeast strains ethyl acetate yield through a precise and seamless genetic manipulation strategy without any extraneous DNA sequences is an essential requisite and significant demand.
Objectives: For increasing the ethyl acetate yield of industrial brewer's yeast strain, all the alleles were overexpressed through "self-cloning" integration strategy.
Material And Methods: strain DH5α was utilized for plasmid construction. alleles were overexpressed through a precise and seamless insertion of the promoter in industrial brewer's yeast strain S6. In addition, growth rates, mRNA levels, AATase activity, the fermentation performance of the engineered strains, and gas chromatography (GC) analysis was conducted.
Results: The two engineered strains (S6-P-12 and S6-P-30) overexpressed all alleles but unaffected normal growth. The mRNA levels of the S6-P-12 and S6-P-30 were all 4-fold higher than that of S6. The AATase (Alcohol acetyl transferases, encoded by gene) activity of the two engineered strains was all 3-fold higher than that of the parent strain. In the beer fermentation at 10 ℃, the concentrations of ethyl acetate produced by the engineered strains S6-P-12 and S6-P-30 was increased to 23.98 and 24.00 mg L, respectively, about 20.44% and 20.54% higher than that of S6.
Conclusions: These results verify that the ethyl acetate yield could be enhanced by the overexpressed of in the polyploid industrial brewer's yeast strains via "self-cloning" integration strategy. The present study provides a reference for target gene modification in the diploid or polyploid industrial yeast strains.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6697848 | PMC |
| http://dx.doi.org/10.21859/ijb.1990 | DOI Listing |
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