scholarly journals Saccharomyces Cerevisiae Strains Selected from Nature Significantly Increased the Production of 2-Phenylethanol Through Protoplast Fusion

2021 ◽  
Author(s):  
Lucheng Lin ◽  
Zhiwei Xu ◽  
Weixia Wang ◽  
Kun Wang ◽  
Tingheng Zhu
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Protoplast Fusion ◽  
Agar Plate ◽  
Breeding Strategy ◽  
Phenotypic Traits ◽  
Industrial Strain ◽  
Genetic Traits ◽  
Good Tolerance ◽  
Yeast Strains ◽  
Thermotolerant Strain

Abstract Background: 2-Phenylethanol (2-PE) is an aromatic alcohol with rose fragrance, which is widely used as an additive in food, tobacco and daily chemical industries. Yeast is the main microorganism producing natural 2-PE, but it is limited by low production and weak tolerance. Nature and fermented products is a resource treasury of yeasts with excellent traits. Screening strains with good phenotypic traits and conducting breeding by cell fusion for genetic pyramiding is an effective way to improve strains. Results: In this study, 25 strains of 2-PE-producing yeasts were isolated from Chinese brewed samples. Three Saccharomyces cerevisiae strains with good traits in tolerance and 2-PE titre were screened out. The strain LSC-1 produces 2-PE of 3.41 g/L with an increase of 9.3% compared to the industrial strain CWY132. The strain NGER shows good tolerance to 2-PE at the concentration of 3.60 g/L in agar plate, and the thermotolerant strain S.C-1 shows growth ability at 41℃. Two rounds of protoplast fusion were performed with these three parent strains for pyramiding of traits. A fusant strain RH2-16 with high 2-PE titre and increased tolerance was obtained. Using 5g/L L-phenylalanine as the precursor substrate, the maximum titre of 2-PE produced by the RH2-16 strain through fermentation and transformation is 4.31 g/L, and the average titre is 4.04 g/L. The molar conversion rate of L-Phe reached 115% in 36 h. Compared to the parental strain LSC-1 and the industrial strain CWY132, 2-PE titre in RH2-16 increased by 26.4% and 38.1%, respectively.Conclusion: Diversified S. cerevisiae strains with different traits can be isolated from the brewing related samples. Protoplast fusion technology can effectively pyramid excellent genetic traits and breed yeast strains with significantly improved tolerance and 2-PE titre. Our research provided a breeding strategy for S. cerevisiae and a strain for industrial production of 2-PE.

scholarly journals Saccharomyces Cerevisiae Strains Selected From Nature Significantly Increased the Production of 2-Phenylethanol Through Protoplast Fusion

2021 ◽  
Author(s):  
Lucheng Lin ◽  
Zhiwei Xu ◽  
Weixia Wang ◽  
Kun Wang ◽  
Tingheng Zhu
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Protoplast Fusion ◽  
Agar Plate ◽  
Breeding Strategy ◽  
Phenotypic Traits ◽  
Industrial Strain ◽  
Genetic Traits ◽  
Fusion Strain ◽  
Thermotolerant Strain ◽  
Key Enzyme

Abstract Background: 2-Phenylethanol (2-PE) is an aromatic alcohol with rose fragrance, which is widely used as an additive in food, tobacco and daily chemical industries. Yeast is the main microorganism producing natural 2-PE, but it is limited by low yield and weak tolerance. Nature and fermented products is a resource treasury of yeasts with excellent traits. Screening strains with good phenotypic traits and conducting breeding by cell fusion for genetic pyramiding is an effective way to improve strains. Results: In this study, 25 strains of 2-PE-producing yeasts were isolated from Chinese brewed samples. Three Saccharomyces cerevisiae strains with good traits in tolerance and 2-PE yield were screened out. The strain LSC-1 produces 2-PE of 3.41 g/L with an increase of 9.3% compared to the industrial strain CWY132. The strain NGER shows good tolerance to 2-PE at the concentration of 3.60 g/L in agar plate, and the thermotolerant strain S.C-1 shows growth ability at 41℃. Two rounds of protoplast fusion were performed with these three parent strains for pyramiding of traits. A fusion strain RH2-16 with high 2-PE yield and increased tolerance was obtained. Using 5 g/L L-phenylalanine as the precursor, RH2-16 produced 2-PE of 4.31 g/L through fermentation conversion and the molar conversion rate of L-Phe reached 115% in 36 h. Compared to the yield of the parental strain LSC-1 and the industrial strain CWY132, 2-PE in RH2-16 increased by 26.4% and 38.1%, respectively. Overexpression of the key enzyme genes ARO8, ARO10, and ADH2 in the Ehrlich pathway in RH2-16 did not increase 2-PE production.Conclusion: Diversified S.cerevisiae strains with different traits can be isolated from the brewing related samples. Protoplast fusion technology can effectively pyramid excellent genetic traits and breed yeast strains with significantly improved tolerance and 2-PE yield. Our research provided a breeding strategy for S.cerevisiae and a strain for industrial production of 2-PE. Overexpression of the key enzyme genes in 2-PE synthesis pathway does not necessarily improve increase production.

Effects of elevated temperature on growth, respiratory-deficient mutation, respiratory activity, and ethanol production in yeast

1988 ◽  
Vol 34 (8) ◽  
pp. 1014-1017 ◽  
Author(s):  
Midori Yamamura ◽  
Yoichi Nagami ◽  
Vitchuporn Vongsuvanlert ◽  
Jaroon Kumnuanta ◽  
Teijiro Kamihara
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Elevated Temperature ◽  
Yeast Extract ◽  
Respiratory Activity ◽  
Culture Conditions ◽  
Inoculum Size ◽  
Yeast Strains ◽  
Thermotolerant Strain ◽  
Grown Culture ◽  
Respiratory Deficient

Some mesophilic yeasts and a thermotolerant strain of Saccharomyces cerevisiae were found to grow at 40 °C in complex media containing 1% yeast extract when an inoculum of 106 or more cells∙mL−1 was used. Yeast extract (6%) permitted Saccharomyces cerevisiae to grow at 40 °C even with a smaller inoculum size (105 cells∙mL−1). The fraction of respiratory-deficient (petite) mutants in 40 °C grown culture was less than 10% except for the thermotolerant strain, which showed greatly increased levels depending on culture conditions. Seven of eight yeast strains exhibited extremely reduced cytochrome oxidase activity when grown at 40 °C irrespective of the frequency of the petite mutation. In contrast, the accumulation of ethanol in the medium and the ethanol-producing activity of the cells were not affected by growth at 40 °C.

Taxonomic characterization and identification of Saccharomyces cerevisiae D8 for brandy production from pineapple

2018 ◽  
Vol 39 (4) ◽  
pp. 474-482
Author(s):  
Hoang Thi Le Thuong ◽  
Nguyen Quang Hao ◽  
Tran Thi Thuy
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Low Ph ◽  
Yeast Strains ◽  
Biological Studies ◽  
Taxonomic Characterization

Eight yeast strains (denoted as D1 to D8) were isolated from samples of natural fermented pineapple. Strain D8 showed highest alcoholic production at low pH and special aroma of pineapple has been chosen for further study. Taxonomic characterization of strain D8 using morphological, biochemical and molecular biological studies confirmed that strain D8  belong to Saccharomycetaceae family, Saccharomycetales order and Saccharomyces cerevisiae species. Therefore, we named this strain as Saccharomyces cerevisiae D8 for further study on Brandy production from pineapple. Citation: Hoang Thi Le Thuong, Nguyen Quang Hao, Tran Thi Thuy, 2017. Taxonomic characterization and identification of Saccharomyces cerevisiae D8 for brandy production from pineapple. Tap chi Sinh hoc, 39(4): 474- 482. DOI: 10.15625/0866-7160/v39n4.10864.*Corresponding author: [email protected] Received 5 December 2016, accepted 12 August 2017

scholarly journals Reciprocal hemizygosity analysis reveals that the Saccharomyces cerevisiae CGI121 gene affects lag time duration in synthetic grape must

2021 ◽  
Author(s):  
Runze Li ◽  
Rebecca C Deed
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Low Temperatures ◽  
Lag Time ◽  
Lag Phase ◽  
Phenotypic Traits ◽  
Time Duration ◽  
Phase Duration ◽  
Grape Must ◽  
The Impact ◽  
Reciprocal Hemizygosity Analysis

Abstract It is standard practice to ferment white wines at low temperatures (10-18 °C). However, low temperatures increase fermentation duration and risk of problem ferments, leading to significant costs. The lag duration at fermentation initiation is heavily impacted by temperature; therefore, identification of Saccharomyces cerevisiae genes influencing fermentation kinetics is of interest for winemaking. We selected 28 S. cerevisiae BY4743 single deletants, from a prior list of open reading frames (ORFs) mapped to quantitative trait loci (QTLs) on chromosomes VII and XIII, influencing the duration of fermentative lag time. Five BY4743 deletants, Δapt1, Δcgi121, Δclb6, Δrps17a, and Δvma21, differed significantly in their fermentative lag duration compared to BY4743 in synthetic grape must (SGM) at 15 °C, over 72 h. Fermentation at 12.5 °C for 528 h confirmed the longer lag times of BY4743 Δcgi121, Δrps17a, and Δvma21. These three candidate ORFs were deleted in S. cerevisiae RM11-1a and S288C to perform single reciprocal hemizygosity analysis (RHA). RHA hybrids and single deletants of RM11-1a and S288C were fermented at 12.5 °C in SGM and lag time measurements confirmed that the S288C allele of CGI121 on chromosome XIII, encoding a component of the EKC/KEOPS complex, increased fermentative lag phase duration. Nucleotide sequences of RM11-1a and S288C CGI121 alleles differed by only one synonymous nucleotide, suggesting that intron splicing, codon bias, or positional effects might be responsible for the impact on lag phase duration. This research demonstrates a new role of CGI121 and highlights the applicability of QTL analysis for investigating complex phenotypic traits in yeast.

scholarly journals Oenological Potential of Autochthonous Saccharomyces cerevisiae Yeast Strains from the Greek Varieties of Agiorgitiko and Moschofilero

Beverages ◽  
2021 ◽  
Vol 7 (2) ◽  
pp. 27
Author(s):  
Dimitrios Kontogiannatos ◽  
Vicky Troianou ◽  
Maria Dimopoulou ◽  
Polydefkis Hatzopoulos ◽  
Yorgos Kotseridis
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Alcoholic Fermentation ◽  
Ethanol Tolerance ◽  
Random Amplified Polymorphic Dna ◽  
Yeast Strains ◽  
Rapd Pcr ◽  
Autochthonous Yeast ◽  
Polymerase Chain ◽  
Grape Varieties ◽  
H2s Production

Nemea and Mantinia are famous wine regions in Greece known for two indigenous grape varieties, Agiorgitiko and Moschofilero, which produce high quality PDO wines. In the present study, indigenous Saccharomyces cerevisiae yeast strains were isolated and identified from spontaneous alcoholic fermentation of Agiorgitiko and Moschofilero musts in order to evaluate their oenological potential. Random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) recovered the presence of five distinct profiles from a total of 430 yeast isolates. The five obtained strains were evaluated at microvinifications trials and tested for basic oenological and biochemical parameters including sulphur dioxide and ethanol tolerance as well as H2S production in sterile grape must. The selected autochthonous yeast strains named, Soi2 (Agiorgitiko wine) and L2M (Moschofilero wine), were evaluated also in industrial (4000L) fermentations to assess their sensorial and oenological characteristics. The volatile compounds of the produced wines were determined by GC-FID. Our results demonstrated the feasibility of using Soi2 and L2M strains in industrial fermentations for Agiorgitiko and Moschofilero grape musts, respectively.

scholarly journals Mating of 2 Laboratory Saccharomyces cerevisiae Strains Resulted in Enhanced Production of 2-Phenylethanol by Biotransformation of L-Phenylalanine

2017 ◽  
Vol 27 (2) ◽  
pp. 81-90 ◽  
Author(s):  
Jolanta Mierzejewska ◽  
Aleksandra Tymoszewska ◽  
Karolina Chreptowicz ◽  
Kamil Krol
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Batch Culture ◽  
Fold Increase ◽  
Biotechnological Production ◽  
Aromatic Alcohol ◽  
Enhanced Production ◽  
Yeast Strains ◽  
First Time

2-Phenylethanol (2-PE) is an aromatic alcohol with a rosy scent which is widely used in the food, fragrance, and cosmetic industries. Promising sources of natural 2-PE are microorganisms, especially yeasts, which can produce 2-PE by biosynthesis and biotransformation. Thus, the first challenging goal in the development of biotechnological production of 2-PE is searching for highly productive yeast strains. In the present work, 5 laboratory <i>Saccharomyces cerevisiae</i> strains were tested for the production of 2-PE. Thereafter, 2 of them were hybridized by a mating procedure and, as a result, a new diploid, <i>S. cerevisiae</i> AM1-d, was selected. Within the 72-h batch culture in a medium containing 5 g/L of <smlcap>L</smlcap>-phenylalanine, AM1-d produced 3.83 g/L of 2-PE in a shaking flask. In this way, we managed to select the diploid <i>S. cerevisiae</i> AM1-d strain, showing a 3- and 5-fold increase in 2-PE production in comparison to parental strains. Remarkably, the enhanced production of 2-PE by the hybrid of 2 yeast laboratory strains is demonstrated here for the first time.

Role of cations in the flocculation of Saccharomyces cerevisiae and discrimination of the corresponding proteins

1991 ◽  
Vol 37 (5) ◽  
pp. 397-403 ◽  
Author(s):  
Hiroshi Kuriyama ◽  
Itaru Umeda ◽  
Harumi Kobayashi
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Cell Surface ◽  
Inhibitory Effect ◽  
Surface Proteins ◽  
Promoting Effect ◽  
Yeast Strains ◽  
Yeast Flocculation ◽  
Different Types ◽  
Anionic Groups

Asexual yeast flocculation was studied using strong flocculents of Saccharomyces cerevisiae. The inhibitory effect of cations on flocculation is considered to be caused by competition between those cations and Ca2+ at the binding site of the Ca2+-requiring protein that is involved in flocculation. Inhibition of flocculation by various cations occurred in the following order: La3+, Sr2+, Ba2+, Mn2+, Al3+, and Na+. Cations such as Mg2+, Co2+, and K+ promoted flocculation. This promoting effect may be based on the reduction of electrostatic repulsive force between cells caused by binding of these cations anionic groups present on the cell surface. In flocculation induced by these cations, trace amounts of Ca2+ excreted on the cell surface may activate the corresponding protein. The ratio of Sr2+/Ca2+ below which cells flocculated varied among strains: for strains having the FLO5 gene, it was 400 to 500; for strains having the FLO1 gene, about 150; and for two alcohol yeast strains, 40 to 50. This suggests that there are several different types of cell surface proteins involved in flocculation in different yeast strains. Key words: yeast, flocculation, protein, cation, calcium.

Mutation in genes coding for glucose-induced degradation-deficient protein contribute to high malate production in yeast strains No. 28 and No. 77 used for industrial brewing of sake

2021 ◽  
Author(s):  
Hiroaki Negoro ◽  
Atsushi Kotaka ◽  
Hiroki Ishida
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Organic Acids ◽  
Nonsense Mutation ◽  
Yeast Strain ◽  
Homozygous Mutation ◽  
Alcohol Fermentation ◽  
Protein Coding ◽  
Protein Coding Genes ◽  
Yeast Strains

ABSTRACT Saccharomyces cerevisiae produces organic acids including malate during alcohol fermentation. Since malate contributes to the pleasant flavor of sake, high-malate-producing yeast strain No. 28 and No. 77 have been developed by the Brewing Society of Japan. In this study, the genes responsible for the high malate phenotype in these strains were investigated. We had found previously that the deletion of components of the glucose induced degradation-deficient (GID) complex led to high malate production in yeast. Upon examining GID protein-coding genes in yeast strain No. 28 and No. 77, a nonsense homozygous mutation of GID4 in strain No. 28, and of GID2 in strain No. 77, were identified as the cause of high malate production. Furthermore, complementary tests of these mutations indicated that the heterozygous nonsense mutation in GID2 was recessive. In contrast, the heterozygous nonsense mutation in GID4 was considered semi-dominant.

Sign in / Sign up

Export Citation Format

Share Document