A Fluorometric Assay for Proteinase a in Beer and Its Application for the Investigation of Enzymatic Effects on Foam Stability

Yokoi, Shigehisa; Shigyo, Tatsuro; Tamaki, Teruo

doi:10.1002/j.2050-0416.1996.tb00892.x

Cited by 17 publications

(8 citation statements)

References 10 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…As a consequence, many researchers have focused on foam-positive species, in particular, beer proteins, which have been implicated to present in the production and stabilization of foam. The decrease in foam stability caused by yeast proteinase A (PrA) in unpasteurized beer has been reported by several researchers (Muldbjerg et al, 1993;Yokoi et al, 1996;Cooper et al, 2000;Wang et al, 2005;He et al, 2006). PrA is coded by PEP4 gene; construction of PEP4 yeast strains would be helpful to keep foam stability (Wang et al, 2007a).…”

Section: Introductionmentioning

confidence: 98%

Construction of recombinant industrial Saccharomyces cerevisiae strain with bglS gene insertion into PEP4 locus by homologous recombination

Zhang

Chen

et al. 2008

J. Zhejiang Univ. Sci. B

View full text Add to dashboard Cite

The bglS gene encoding endo-l,3-1,4-beta-glucanase from Bacillus subtilis was cloned and sequenced in this study. The bglS expression cassette, including PGK1 promoter, bglS gene fused to the signal sequence of the yeast mating pheromone alpha-factor (MFalpha1(S)), and ADH1 terminator with G418-resistance as the selected marker, was constructed. Then one of the PEP4 allele of Saccharomyces cerevisiae WZ65 strain was replaced by bglS expression cassette using chromosomal integration of polymerase chain reaction (PCR)-mediated homologous recombination, and the bglS gene was expressed simultaneously. The recombinant strain S. cerevisiae (SC-betaG) was preliminarily screened by the clearing hydrolysis zone formed after the barley beta-glucan was hydrolyzed in the plate and no proteinase A (PrA) activity was measured in fermenting liquor. The results of PCR analysis of genome DNA showed that one of the PEP4 allele had been replaced and bglS gene had been inserted into the locus of PEP4 gene in recombinant strains. Different endo-l,3-1,4-beta-glucanase assay methods showed that the recombinant strain SC-betaG had high endo-l,3-1,4-beta-glucanase expression level with the maximum of 69.3 U/(h.ml) after 60 h of incubation. Meanwhile, the Congo Red method was suitable for the determination of endo-l,3-1,4-beta-glucanase activity during the actual brewing process. The current research implies that the constructed yeast strain could be utilized to improve the industrial brewing property of beer.

show abstract

Section: Introductionmentioning

confidence: 98%

Construction of recombinant industrial Saccharomyces cerevisiae strain with bglS gene insertion into PEP4 locus by homologous recombination

Zhang

Chen

et al. 2008

J. Zhejiang Univ. Sci. B

View full text Add to dashboard Cite

show abstract

“…, 1980; Magni et al. , 1982; Yokoi et al. , 1996), but it was more difficult to purify the enzyme from yeast than from fresh beer because there were a lot of proteins similar to proteinase A in molecular size, and the enzyme activity was easily lost during the operations of chromatography (data and photograph were not shown).…”

Section: Resultsmentioning

confidence: 99%

“…Proteinase A can be excreted into beer under stress or by yeast autolysis (Maddox & Hough, 1970) and is detrimental to beer foam stability (Dreyer et al. , 1983; Yokoi et al. , 1996).…”

Section: Introductionmentioning

confidence: 99%

“…Yeast proteinase A (PrA) [EC3.4.23.25], a proteolytic enzyme found in the vacuole of Saccharomyces cerevisiae, is involved in non-specific proteolysis during starvation and proteolytic maturation of other vacuolar enzymes (Woodford et al, 1993;Rupp & Wolf, 1995;van den Hazel et al, 1995;Westphal et al, 1996). Proteinase A can be excreted into beer under stress or by yeast autolysis (Maddox & Hough, 1970) and is detrimental to beer foam stability (Dreyer et al, 1983;Yokoi et al, 1996). The enzyme has been intensively investigated on its structure (Dreyer et al, 1986;Aguilar et al, 1997;Gustchina et al, 2002), function (Jones, 1977(Jones, , 1991Jones et al, 1982;Ammerer et al, 1986;Harris & Cotter, 1987;Teichert et al, 1989;Rupp et al, 1991;Hazel et al, 1992) and enzymatic characteristics (Hata et al, 1967a;Meussdoerffer et al, 1980;Magni et al, 1982), etc.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Purification of yeast proteinase A from fresh beer and its specificity on foam proteins

Wang

Liu

et al. 2005

Int J of Food Sci Tech

View full text Add to dashboard Cite

Yeast proteinase A from fresh beer was first purified with Sephadex G-100 column chromatography and the active fractions reached to 5.3-fold purification with 7% of yield. After purification with DEAE Sephadex A50, proteinase A activity increased to be 10.1 times of the initial with 1% of yield. When identifying the sample from chromatography by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), only one protein band with 42 kDa was observed, this indicated that the enzyme was purified. The pattern of electrophoresis of hydrolysed beer by crude proteinase A did not show lipid transfer protein (LTP) on the gel. The result of SDS-PAGE of interaction mixture of purified proteinase A and beer also indicated that LTP was decomposed.

show abstract

“…Many experiments have shown that this phenomenon is due mainly to Proteinase A degradation of beer proteins 12 . To maintain the stability of beer foam, a number of studies have been conducted on Proteinase A to identify the relationship between Proteinase A content and foam stability 1,4,20,24,26 .…”

Section: Introductionmentioning

confidence: 99%

Production of Beer with a Genetically Engineered Strain of S. cerevisiae with Modified Beta Glucanase Expression

Yang¹,

Shengzhou²,

He³

et al. 2009

Journal of the Institute of Brewing

View full text Add to dashboard Cite

This study used a recombinant Saccharomyces cerevisiae strain, which expressed both β-glucanase enzyme and reduced Proteinase A expression during wort fermentations. The genetic stability and fermentation features of the strain were examined. The recombinant strain's proteinase A activity was reduced compared to the parent strain; β-glucanase was produced throughout the fermentation. The fermentation with the recombinant S. cerevisiae strain exhibited a larger reduction in β-glucan content than what was observed with the control strain, with β-glucan degradation above 80%. The foam stability period was reduced when the beer produced by the recombinant S. cerevisiae was stored for 3 months. SDS-PAGE analysis of the beer proteins indicated that lipid transfer protein 1 had disappeared. Fermentation studies indicated that based on the parameters examined, this recombinant strain was suitable for industrial beer production.

show abstract

A Fluorometric Assay for Proteinase a in Beer and Its Application for the Investigation of Enzymatic Effects on Foam Stability

Cited by 17 publications

References 10 publications

Construction of recombinant industrial Saccharomyces cerevisiae strain with bglS gene insertion into PEP4 locus by homologous recombination

Construction of recombinant industrial Saccharomyces cerevisiae strain with bglS gene insertion into PEP4 locus by homologous recombination

Purification of yeast proteinase A from fresh beer and its specificity on foam proteins

Production of Beer with a Genetically Engineered Strain of S. cerevisiae with Modified Beta Glucanase Expression

Contact Info

Product

Resources

About