scholarly journals Metabolic Engineering of Saccharomyces cerevisiae for Industrial Biotechnology

2021 ◽  
Author(s):  
Seyma Hande Tekarslan-Sahin
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Metabolic Engineering ◽  
Genetic Engineering ◽  
Value Added ◽  
Industrial Biotechnology ◽  
Biotechnological Applications ◽  
Pharmaceutical Ingredients ◽  
Yeast Strains ◽  
Physiology And Biochemistry ◽  
Industrial Strains

Saccharomyces cerevisiae is an important and popular host for production of value-added molecules such as pharmaceutical ingredients, therapeutic proteins, chemicals, biofuels and enzymes. S. cerevisiae, the baker’s yeast, is the most used yeast model as there is an abundance of knowledge on its genetics, physiology and biochemistry, and also it has numerous applications in genetic engineering and fermentation technologies. There has been an increasing interest in developing and improving yeast strains for industrial biotechnology. Metabolic engineering is a tool to develop industrial strains by manipulating yeast metabolism to enhance the production of value-added molecules. This chapter reviews the metabolic engineering strategies for developing industrial yeast strains for biotechnological applications and highlights recent advances in this field such as the use of CRISPR/Cas9.

scholarly journals Progress in Metabolic Engineering of Saccharomyces cerevisiae

2008 ◽  
Vol 72 (3) ◽  
pp. 379-412 ◽  
Author(s):  
Elke Nevoigt
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Metabolic Engineering ◽  
Strain Improvement ◽  
Fine Tuning ◽  
Industrial Biotechnology ◽  
Evolutionary Engineering ◽  
Traditional Use ◽  
Inverse Metabolic Engineering ◽  
Physiology And Biochemistry ◽  
High Ethanol

SUMMARY The traditional use of the yeast Saccharomyces cerevisiae in alcoholic fermentation has, over time, resulted in substantial accumulated knowledge concerning genetics, physiology, and biochemistry as well as genetic engineering and fermentation technologies. S. cerevisiae has become a platform organism for developing metabolic engineering strategies, methods, and tools. The current review discusses the relevance of several engineering strategies, such as rational and inverse metabolic engineering, evolutionary engineering, and global transcription machinery engineering, in yeast strain improvement. It also summarizes existing tools for fine-tuning and regulating enzyme activities and thus metabolic pathways. Recent examples of yeast metabolic engineering for food, beverage, and industrial biotechnology (bioethanol and bulk and fine chemicals) follow. S. cerevisiae currently enjoys increasing popularity as a production organism in industrial (“white”) biotechnology due to its inherent tolerance of low pH values and high ethanol and inhibitor concentrations and its ability to grow anaerobically. Attention is paid to utilizing lignocellulosic biomass as a potential substrate.

scholarly journals Yeast-Based Biosynthesis of Natural Products From Xylose

2021 ◽  
Vol 9 ◽  
Author(s):  
Jian Zha ◽  
Miaomiao Yuwen ◽  
Weidong Qian ◽  
Xia Wu
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Natural Products ◽  
Metabolic Engineering ◽  
State Of The Art ◽  
Commercial Production ◽  
Scheffersomyces Stipitis ◽  
Lignocellulosic Hydrolysates ◽  
Yeast Strains ◽  
Future Challenges ◽  
Biomass Materials

Xylose is the second most abundant sugar in lignocellulosic hydrolysates. Transformation of xylose into valuable chemicals, such as plant natural products, is a feasible and sustainable route to industrializing biorefinery of biomass materials. Yeast strains, including Saccharomyces cerevisiae, Scheffersomyces stipitis, and Yarrowia lipolytica, display some paramount advantages in expressing heterologous enzymes and pathways from various sources and have been engineered extensively to produce natural products. In this review, we summarize the advances in the development of metabolically engineered yeasts to produce natural products from xylose, including aromatics, terpenoids, and flavonoids. The state-of-the-art metabolic engineering strategies and representative examples are reviewed. Future challenges and perspectives are also discussed on yeast engineering for commercial production of natural products using xylose as feedstocks.

scholarly journals Rapid Colorimetric Detection of Genome Evolution in SCRaMbLEd Synthetic Saccharomyces cerevisiae Strains

2020 ◽  
Vol 8 (12) ◽  
pp. 1914
Author(s):  
Elizabeth L. I. Wightman ◽  
Heinrich Kroukamp ◽  
Isak S. Pretorius ◽  
Ian T. Paulsen ◽  
Helena K. M. Nevalainen
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Genome Evolution ◽  
Colorimetric Detection ◽  
Cre Recombinase ◽  
Control Measures ◽  
Genomic Rearrangements ◽  
Industrial Yeast ◽  
Poor Growth ◽  
Yeast Strains ◽  
Industrial Strains

Genome-scale engineering and custom synthetic genomes are reshaping the next generation of industrial yeast strains. The Cre-recombinase-mediated chromosomal rearrangement mechanism of designer synthetic Saccharomyces cerevisiae chromosomes, known as SCRaMbLE, is a powerful tool which allows rapid genome evolution upon command. This system is able to generate millions of novel genomes with potential valuable phenotypes, but the excessive loss of essential genes often results in poor growth or even the death of cells with useful phenotypes. In this study we expanded the versatility of SCRaMbLE to industrial strains, and evaluated different control measures to optimize genomic rearrangement, whilst limiting cell death. To achieve this, we have developed RED (rapid evolution detection), a simple colorimetric plate-assay procedure to rapidly quantify the degree of genomic rearrangements within a post-SCRaMbLE yeast population. RED-enabled semi-synthetic strains were mated with the haploid progeny of industrial yeast strains to produce stress-tolerant heterozygous diploid strains. Analysis of these heterozygous strains with the RED-assay, genome sequencing and custom bioinformatics scripts demonstrated a correlation between RED-assay frequencies and physical genomic rearrangements. Here we show that RED is a fast and effective method to evaluate the optimal SCRaMbLE induction times of different Cre-recombinase expression systems for the development of industrial strains.

scholarly journals A teaching protocol demonstrating the use of EasyClone and CRISPR/Cas9 for metabolic engineering of Saccharomyces cerevisiae and Yarrowia lipolytica

2019 ◽  
Author(s):  
N Milne ◽  
L R R Tramontin ◽  
I Borodina
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Metabolic Engineering ◽  
Yarrowia Lipolytica ◽  
Yeast Species ◽  
Production Capacity ◽  
Value Added ◽  
Phenotypic Screen ◽  
Rate Limiting ◽  
Β Carotene

ABSTRACT We present a teaching protocol suitable for demonstrating the use of EasyClone and CRISPR/Cas9 for metabolic engineering of industrially relevant yeasts Saccharomyces cerevisiae and Yarrowia lipolytica, using β-carotene production as a case study. The protocol details all steps required to generate DNA parts, transform and genotype yeast, and perform a phenotypic screen to determine β-carotene production. The protocol is intended to be used as an instruction manual for a two-week practical course aimed at MSc and PhD students. The protocol details all necessary steps for students to engineer yeast to produce β-carotene and serves as a practical introduction to the principles of metabolic engineering including the concepts of boosting native precursor supply and alleviating rate-limiting steps. It also highlights key differences in the metabolism and heterologous production capacity of two industrially relevant yeast species. The protocol is divided into daily experiments covering a two week period and provides detailed instructions for every step meaning this protocol can be used ‘as is’ for a teaching course or as a case study for how yeast can be engineered to produce value-added molecules.

Inability of Petite Mutants of Industrial Yeasts to Utilize Various Sugars, and a Comparison with the Ability of the Parent Strains to Ferment the Same Sugars Microaerophilically

1983 ◽  
Vol 38 (5-6) ◽  
pp. 405-407 ◽  
Author(s):  
J. F. T. Spencer ◽  
D. M. Spencer ◽  
R. Miller
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Methyl Glucoside ◽  
Yeast Strains ◽  
Petite Mutants ◽  
Industrial Strains ◽  
Industrial Yeasts ◽  
Microaerophilic Conditions

A number of industrial strains of Saccharomyces cerevisiae were converted to the petite form and tested for the ability to utilize galactose, maltose, sucrose, α-methyl glucoside and raffinose. The parent strains all metabolized these sugars aerobically. Twelve of the petite forms did not utilize galactose, six failed to utilize maltose, 17 did not utilize x-methyl glucoside, and 18 did not utilize raffinose. The petites of two distiller’s yeast strains did not utilize sucrose. The respiratory-competent parent strains nearly all fermented galactose, maltose, sucrose and raffinose, though 19 strains did not ferment α-methyl glucoside microaerophilically. Three strains did not ferment galactose, two fermented it only after several days adaptation, one did not ferment raffinose, and two did not ferment sucrose under microaerophilic conditions. Six respiratory-competent strains which did not utilize galactose when in the petite form fermented higher (10%) concentrations of glucose and maltose under microaerophilic conditions, but only three of these fermented galactose. The implications of these findings for the use of such strains in industry are discussed briefly.

scholarly journals Adaptive Evolution of Industrial Brewer’s Yeast Strains towards a Snowflake Phenotype

Fermentation ◽  
2020 ◽  
Vol 6 (1) ◽  
pp. 20 ◽  
Author(s):  
Yeseren Kayacan ◽  
Thijs Van Mieghem ◽  
Filip Delvaux ◽  
Freddy R. Delvaux ◽  
Ronnie Willaert
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Cell Aggregation ◽  
Adaptive Laboratory Evolution ◽  
Laboratory Evolution ◽  
Sedimentation Behavior ◽  
Yeast Strains ◽  
Industrial Strains ◽  
Genetic Interference ◽  
Cost Efficient ◽  
Industrial Brewer’S Yeast

Flocculation or cell aggregation is a well-appreciated characteristic of industrial brewer’s strains, since it allows removal of the cells from the beer in a cost-efficient and environmentally-friendly manner. However, many industrial strains are non-flocculent and genetic interference to increase the flocculation characteristics are not appreciated by the consumers. We applied adaptive laboratory evolution (ALE) to three non-flocculent, industrial Saccharomyces cerevisiae brewer’s strains using small continuous bioreactors (ministats) to obtain an aggregative phenotype, i.e., the “snowflake” phenotype. These aggregates could increase yeast sedimentation considerably. We evaluated the performance of these evolved strains and their produced flavor during lab scale beer fermentations. The small aggregates did not result in a premature sedimentation during the fermentation and did not result in major flavor changes of the produced beer. These results show that ALE could be used to increase the sedimentation behavior of non-flocculent brewer’s strains.

scholarly journals Saccharomyces Cerevisiae—An Interesting Producer of Bioactive Plant Polyphenolic Metabolites

2020 ◽  
Vol 21 (19) ◽  
pp. 7343
Author(s):  
Grzegorz Chrzanowski
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Defense Mechanisms ◽  
Chemical Diversity ◽  
Industrial Biotechnology ◽  
Host Organism ◽  
Biotechnological Applications ◽  
Phenolic Metabolites ◽  
Cell Factories ◽  
Plant Genes ◽  
Agricultural Industries

Secondary phenolic metabolites are defined as valuable natural products synthesized by different organisms that are not essential for growth and development. These compounds play an essential role in plant defense mechanisms and an important role in the pharmaceutical, cosmetics, food, and agricultural industries. Despite the vast chemical diversity of natural compounds, their content in plants is very low, and, as a consequence, this eliminates the possibility of the production of these interesting secondary metabolites from plants. Therefore, microorganisms are widely used as cell factories by industrial biotechnology, in the production of different non-native compounds. Among microorganisms commonly used in biotechnological applications, yeast are a prominent host for the diverse secondary metabolite biosynthetic pathways. Saccharomyces cerevisiae is often regarded as a better host organism for the heterologous production of phenolic compounds, particularly if the expression of different plant genes is necessary.

scholarly journals Saccharomyces Cerevisiae an Interesting Producer of Bioactive Plant Polyphenolic Metabolites

2020 ◽  
Author(s):  
Grzegorz Chrzanowski
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Defense Mechanisms ◽  
Chemical Diversity ◽  
Industrial Biotechnology ◽  
Host Organism ◽  
Biotechnological Applications ◽  
Phenolic Metabolites ◽  
Cell Factories ◽  
Plant Genes ◽  
Agricultural Industries

Secondary phenolic metabolites are defined as valuable natural products synthesized by different organisms that are not essential for growth and development. These compounds play an essential role in plant defense mechanisms, and an important role in the pharmaceutical, cosmetics, food, and agricultural industries. Despite the vast chemical diversity of natural compounds, their content in plants is very low, in consequence, it eliminates the possibility of the production of these interesting secondary metabolites from plants. Therefore, microorganisms are widely used as cell factories by industrial biotechnology to the production of different non-native compounds. Among microorganisms commonly used in biotechnological applications, yeasts are prominent host for the diverse secondary metabolite biosynthetic pathways. Saccharomyces cerevisiae is often regarded as the better host organism for the heterologous production of phenolics compounds, especially if the expression of different plant genes is necessary.

scholarly journals Rapid Colorimetric Detection of Genome Evolution in SCRaMbLEd Synthetic Saccharomyces cerevisiae Strains

2020 ◽  
Author(s):  
Elizabeth L. I. Wightman ◽  
Heinrich Kroukamp ◽  
Isak S. Pretorius ◽  
Ian T. Paulsen ◽  
Helena K. M. Nevalainen
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Genome Evolution ◽  
Colorimetric Detection ◽  
Rapid Evolution ◽  
Control Measures ◽  
Genomic Rearrangements ◽  
Industrial Yeast ◽  
Poor Growth ◽  
Yeast Strains ◽  
Industrial Strains

Genome-scale engineering and custom synthetic genomes are reshaping the next generation of industrial yeast strains. The Cre-recombinase mediated chromosomal rearrangement mechanism of designer synthetic Saccharomyces cerevisiae chromosomes, known as SCRaMbLE, is a powerful tool which allows rapid genome evolution upon command. This system is able to generate millions of novel genomes with potential valuable phenotypes, but the excessive loss of essential genes often results in poor growth or even the death of cells with useful phenotypes. In this study we expanded the versatility of SCRaMbLE to industrial strains, and evaluated different control measures to optimise genomic rearrangement, whilst limiting cell death. To achieve this, we have developed RED (Rapid Evolution Detection), a simple colorimetric plate-assay procedure to rapidly quantify the degree of genomic rearrangements within a post-SCRaMbLE yeast population. RED-enabled semi-synthetic strains were mated with haploid progeny of industrial yeast strains to produce stress tolerant heterozygous diploid strains. Analysis of these heterozygous strains with the RED-assay, genome sequencing and custom bioinformatics scripts demonstrated a correlation between RED-assay frequencies and physical genomic rearrangements. Here we show that RED is a fast and effective method to evaluate optimal SCRaMbLE induction times of different Cre-recombinse expression systems for the development of industrial strains.

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