Preface

Yeasts are one of the most important eukaryotic microorganisms due to their role as model organisms for eukaryotic biological studies with a focus on understanding and interpreting human DNA. Recently, researches have been working with several yeast genera and species such as *Saccharomyces cerevisiae*, *Pichia pastoris*, and *Schizosaccharomyces pombe* as model organisms for cancer biology investigation, including tumorigenic mechanism studies, development, and production of anticancer drugs.

Many thousands of years ago, Egyptian, Babylonian, and Greek civilizations used these microorganisms to produce fermented foods and beverages, such as bread, beer, and wine. Since then, yeasts have been applied to the food industry and their utilization has been increasing over the last few decades. On the other hand, new yeast platforms such as cell factories have been developed for the production of biochemicals such as fuels and drugs.

This book offers a broad understanding of yeast applications such as cell factories for the production of biofuel, food, and bioproducts with high value. Additionally, the book provides an overview of yeast utilization as probiotics in animal nutrition. The book starts with an introduction chapter on the advances of yeast utilization in human history up to current biotechnology approaches.

The six chapters are grouped into three sections: Animal Nutrition, Food Industry, and Industrial Bioproducts. The Animal Nutrition section comprises two chapters dealing with the application of yeast as a probiotic in animal nutrition. In these chapters, the reader is guided through the action mechanisms of indigenous and commercial *S. cerevisiae* yeasts on ruminant gut microbial balance and can identify the ideal probiotic yeast for animal feeding.

In the Food Industry section, the advantages of nonconventional yeast strains on the food and beverage industry such as wine making, brewing, coffee and cocoa fermentation, and xylitol production are presented. It also covers the biotransformation process and analytical procedures, such as mechanisms of accumulation, extraction, and quantification of organically bound selenium by *S. cerevisiae.*

The Industrial Bioproducts section deals with the application of yeast as a platform for the biosynthesis of biochemicals such as flavonoids, alkaloids, terpenoids, and saponins used in the agriculture industry. It also covers the application of *Rhodotorula/Rhodosporidium* as a platform cell factory for biofuel feedstock, carotenoids, enzymes, and biosurfactant production.

This book intends to present a broad understanding of several biotechnology applications of yeasts in industrial bioprocesses. It will cover a multitude of issues such as the development of a platform cell factory, the use of nonconventional yeasts, and the diverse application of yeasts on bioprocesses. Other aspects related

**II**

**Section 4**

for Industrial Bioproducts

Industrial Bioproducts **83**

**Chapter 6 85**

**Chapter 7 105**

The Oleaginous Red Yeast *Rhodotorula/Rhodosporidium*: A Factory

*De Novo* Synthesis of Plant Natural Products in Yeast

*by Wentao Sun, Yu-jia Zhao and Chun Li*

*by Mathew Lyman, Salustra Urbin, Cheryl Strout and Bonnee Rubinfeld*

to bioprocesses will also be analyzed, such as biofuels and bioproduct production and yeast application in wine making and brewing.

> **Thalita Peixoto Basso, Ph.D.** University of Sao Paulo, Brazil

> > **1**

Section 1

Introduction

Section 1 Introduction

**IV**

to bioprocesses will also be analyzed, such as biofuels and bioproduct production

**Thalita Peixoto Basso, Ph.D.** University of Sao Paulo,

Brazil

and yeast application in wine making and brewing.

**3**

**Chapter 1**

**1. Introduction**

tion of anticancer drugs [3].

**2. Yeast application**

Biotechnology

*and Carlos Alberto Labate*

*Thalita Peixoto Basso, Luiz Carlos Basso*

fermentation in the final of nineteenth century.

Introductory Chapter: Yeasts in

Yeasts are very important for many reasons. These microorganisms were the first species to be domesticated by man, although not intentionally. For millennia they were used in fermented beverages and foods without knowing their existence. Biochemistry as a science was born when physiologists looked deeper in sugar

Today yeast takes a place in several fields of science and technology. As long as yeast genes and mammal cells encode very similar proteins, these microorganisms are useful as a model to understand and interpret human DNA sequences. Indeed, yeast genetic manipulation is much easier and cheaper than mammalian systems. So

Particularly, *Saccharomyces cerevisiae* is a model organism to study epigenetic traits that can be characterized as a stably heritable phenotype resulting from changes in a chromosome without alteration in the DNA sequence. As a result of yeast small eukaryotic genome, short generation time and easy genetic manipulation [4]. Additionally yeasts are very important players in many economical relevant bioprocessing as bakery, brewery, distilling, food industry, and biofuel, leading yeasts to be considered the most explored and studied eukaryotic microorganism.

Since 8000 years ago in our history, humans have been using microorganisms to produce fermented foods and beverages. More recently chemicals and fuels have been produced by bioprocesses. The development of cell factories has been incentivized for the industrial production of new chemicals. However the development of new yeast platform cell factory is costly and time-consuming. The difficulty to develop new cell factories to produce a specific metabolite is due to metabolism which has evolved to allow cell growth and maintenance to keep homeostasis [5]. Yeasts from phyla of ascomycetes and basidiomycetes have diverse biotechnological application on food industry. They are responsible for a wide range of fermented products such as alcoholic beverages (e.g., beer, wine, and "cachaça"), fermented milk, cheese, bread, and so on. Yeast also has an application in the functional food industry as probiotics and nutraceutical products [6].

yeast has turned out to be a useful model for eukaryotic biology [1, 2].

Furthermore yeasts such as *Saccharomyces cerevisiae*, *Picchia pastoris*, and *Schizosaccharomyces pombe* have been used as model organisms to study cancer biology, including research and development of tumorigenic mechanisms and produc-

#### **Chapter 1**

## Introductory Chapter: Yeasts in Biotechnology

*Thalita Peixoto Basso, Luiz Carlos Basso and Carlos Alberto Labate*

#### **1. Introduction**

Yeasts are very important for many reasons. These microorganisms were the first species to be domesticated by man, although not intentionally. For millennia they were used in fermented beverages and foods without knowing their existence. Biochemistry as a science was born when physiologists looked deeper in sugar fermentation in the final of nineteenth century.

Today yeast takes a place in several fields of science and technology. As long as yeast genes and mammal cells encode very similar proteins, these microorganisms are useful as a model to understand and interpret human DNA sequences. Indeed, yeast genetic manipulation is much easier and cheaper than mammalian systems. So yeast has turned out to be a useful model for eukaryotic biology [1, 2].

Furthermore yeasts such as *Saccharomyces cerevisiae*, *Picchia pastoris*, and *Schizosaccharomyces pombe* have been used as model organisms to study cancer biology, including research and development of tumorigenic mechanisms and production of anticancer drugs [3].

Particularly, *Saccharomyces cerevisiae* is a model organism to study epigenetic traits that can be characterized as a stably heritable phenotype resulting from changes in a chromosome without alteration in the DNA sequence. As a result of yeast small eukaryotic genome, short generation time and easy genetic manipulation [4].

Additionally yeasts are very important players in many economical relevant bioprocessing as bakery, brewery, distilling, food industry, and biofuel, leading yeasts to be considered the most explored and studied eukaryotic microorganism.

#### **2. Yeast application**

Since 8000 years ago in our history, humans have been using microorganisms to produce fermented foods and beverages. More recently chemicals and fuels have been produced by bioprocesses. The development of cell factories has been incentivized for the industrial production of new chemicals. However the development of new yeast platform cell factory is costly and time-consuming. The difficulty to develop new cell factories to produce a specific metabolite is due to metabolism which has evolved to allow cell growth and maintenance to keep homeostasis [5].

Yeasts from phyla of ascomycetes and basidiomycetes have diverse biotechnological application on food industry. They are responsible for a wide range of fermented products such as alcoholic beverages (e.g., beer, wine, and "cachaça"), fermented milk, cheese, bread, and so on. Yeast also has an application in the functional food industry as probiotics and nutraceutical products [6].

*Saccharomyces cerevisiae* has been metabolically engineered for the production of first-generation and second-generation bioethanols, advanced biofuels, and chemicals [7–11].

Recently, new tools for genome editing as CRISPR-Cas9 technology have the advantage to allow introduction of many genes into any chromosome location [12, 13]. On the other hand, high-throughput methods as transcriptomic, proteomic, and metabolomic analyses support the introduction of metabolic pathway over cellular physiology metabolism. Indeed next-generation sequencing allows the identification of any genome modification responsible for desirable phenotype [5].

#### **3. Conclusion**

In conclusion, we believe that the yeasts are a nearly ideal model system for eukaryotic biology at the cellular and molecular level. Additionally their use in increasingly technological applications will augment the importance of yeast for human well-being.

#### **Acknowledgements**

The authors would like to acknowledge CAPES and Inter-unit Bioenergy Post Graduation Program (USP/UNESP/UNICAMP) for PNPD/CAPES postdoctoral fellowship (process number 88882.317582/2019-01), Max Feffer Laboratory from Genetic Department (ESALQ/USP), and Yeast Physiology and Fermentation Laboratory from Biological Science (ESALQ/USP).

#### **Conflict of interest**

The authors confirm there is no conflict of interest.

#### **Author details**

Thalita Peixoto Basso\*, Luiz Carlos Basso and Carlos Alberto Labate "Luiz de Queiroz" College of Agriculture, University of Sao Paulo, Piracicaba, SP, Brazil

\*Address all correspondence to: tpbasso@usp.br

© 2019 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

**5**

*Introductory Chapter: Yeasts in Biotechnology DOI: http://dx.doi.org/10.5772/intechopen.85898*

[1] Botstein D, Fink GR. Yeast: An experimental organism for modern biology. Science. 1988;**240**:1439-1443.

[2] Botstein D, Chervitz SA, Cherry JM. Yeast as a model organism. Science. 1997;**277**:1259-1260. DOI: 10.1116/

[9] Basso TP. Improvement of

for increased tolerance towards inhibitors from second-generation ethanol substrate [thesis]. Piracicaba-SP:

University of Sao Paulo; 2015

10.1016/j.cbpa.2017.10.025

[10] Jin YS, Cate JHD. Metabolic engineering of yeast for lignocellulosic biofuel production. Current Opinion in Chemical Biology. 2017;**41**:99-106. DOI:

[11] Kim SR, Skerker JM, Kong II, Kim H, Maurer MJ, Zhang GC, et al. Metabolic engineering of a haploid strain derived from a triploid industrial yeast for producing cellulosic ethanol. Metabolic Engineering. 2017;**40**:176-185. DOI: 10.1016/j.ymben.2017.02.006

[12] Estrela R, Cate JHD. Energy biotechnology in the CRISPR-Cas9 era. Current Opinion in Biotechnology.

2016;**38**:79-84. DOI: 10.1016/j.

[13] Jakociunas T, Jensen MK, Keasling JD. CRISPR/Cas9 adbances engineering of microbial cell factories. Metabolic Engineering. 2016;**34**:44-59. DOI: 10.1016/j.ymben.2015.12.003

copbio.2016.01.005

*Saccharomyces cerevisiae* by hybridization

[3] Ferreira R, Limeta A, Nielsen J. Tackling cancer with yeast-based technologies. Trends in Biotechnology.

2018;**1735**:1-12. DOI: 10.1016/j.

[4] Fuchs SM, Quasem I. Budding yeast as a model study epigenetics. Drug Discovery Today: Disease Models. 2014;**12**:1-16. DOI: 10.1016/j.

[5] Nielsen J, Keasling JD. Engineering

2016;**164**:1185-1197. DOI: 10.1016/j.

[6] Rai AK, Pandey A, Sahoo D. Biotechnological potential of yeasts in functional food industry. Trends in Food Science and Technology. 2019;**83**:129-137. DOI: 10.1016/j.

[7] Zhou H, Cheng JS, Wang BL, Fink GR, Stephanopoulos G. Xylose isomerase overexpression along with engineering of the pentose phosphate pathway and evolutionary engineering enable rapid xylose utilization and ethanol production by *Saccharomyces cerevisiae*. Metabolic Engineering. 2012;**14**:611-622. DOI: 10.1016/j.

[8] Nielsen J, Larsson C, van Maris A, Pronk J. Metabolic engineering of yeast for production of fuels and chemicals. Current Opinion in Biotechnology. 2013;**24**:398-404. DOI: 10.1016/j.

DOI: 10.1126/science.3287619

science.277.5330.1259

tibtech.2018.11.013

ddmod.2014.04.004

cell.2016.02.004

tifs.2018.11.016

ymben.2012.07.011

copbio.2013.03.023

cellular metabolism. Cell.

**References**

*Introductory Chapter: Yeasts in Biotechnology DOI: http://dx.doi.org/10.5772/intechopen.85898*

### **References**

*Yeasts in Biotechnology*

chemicals [7–11].

able phenotype [5].

**3. Conclusion**

human well-being.

**Acknowledgements**

**Conflict of interest**

**Author details**

Brazil

Laboratory from Biological Science (ESALQ/USP).

The authors confirm there is no conflict of interest.

Thalita Peixoto Basso\*, Luiz Carlos Basso and Carlos Alberto Labate

**4**

provided the original work is properly cited.

\*Address all correspondence to: tpbasso@usp.br

© 2019 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium,

"Luiz de Queiroz" College of Agriculture, University of Sao Paulo, Piracicaba, SP,

*Saccharomyces cerevisiae* has been metabolically engineered for the production of first-generation and second-generation bioethanols, advanced biofuels, and

Recently, new tools for genome editing as CRISPR-Cas9 technology have the advantage to allow introduction of many genes into any chromosome location [12, 13]. On the other hand, high-throughput methods as transcriptomic, proteomic, and metabolomic analyses support the introduction of metabolic pathway over cellular physiology metabolism. Indeed next-generation sequencing allows the identification of any genome modification responsible for desir-

In conclusion, we believe that the yeasts are a nearly ideal model system for eukaryotic biology at the cellular and molecular level. Additionally their use in increasingly technological applications will augment the importance of yeast for

The authors would like to acknowledge CAPES and Inter-unit Bioenergy Post Graduation Program (USP/UNESP/UNICAMP) for PNPD/CAPES postdoctoral fellowship (process number 88882.317582/2019-01), Max Feffer Laboratory from Genetic Department (ESALQ/USP), and Yeast Physiology and Fermentation

[1] Botstein D, Fink GR. Yeast: An experimental organism for modern biology. Science. 1988;**240**:1439-1443. DOI: 10.1126/science.3287619

[2] Botstein D, Chervitz SA, Cherry JM. Yeast as a model organism. Science. 1997;**277**:1259-1260. DOI: 10.1116/ science.277.5330.1259

[3] Ferreira R, Limeta A, Nielsen J. Tackling cancer with yeast-based technologies. Trends in Biotechnology. 2018;**1735**:1-12. DOI: 10.1016/j. tibtech.2018.11.013

[4] Fuchs SM, Quasem I. Budding yeast as a model study epigenetics. Drug Discovery Today: Disease Models. 2014;**12**:1-16. DOI: 10.1016/j. ddmod.2014.04.004

[5] Nielsen J, Keasling JD. Engineering cellular metabolism. Cell. 2016;**164**:1185-1197. DOI: 10.1016/j. cell.2016.02.004

[6] Rai AK, Pandey A, Sahoo D. Biotechnological potential of yeasts in functional food industry. Trends in Food Science and Technology. 2019;**83**:129-137. DOI: 10.1016/j. tifs.2018.11.016

[7] Zhou H, Cheng JS, Wang BL, Fink GR, Stephanopoulos G. Xylose isomerase overexpression along with engineering of the pentose phosphate pathway and evolutionary engineering enable rapid xylose utilization and ethanol production by *Saccharomyces cerevisiae*. Metabolic Engineering. 2012;**14**:611-622. DOI: 10.1016/j. ymben.2012.07.011

[8] Nielsen J, Larsson C, van Maris A, Pronk J. Metabolic engineering of yeast for production of fuels and chemicals. Current Opinion in Biotechnology. 2013;**24**:398-404. DOI: 10.1016/j. copbio.2013.03.023

[9] Basso TP. Improvement of *Saccharomyces cerevisiae* by hybridization for increased tolerance towards inhibitors from second-generation ethanol substrate [thesis]. Piracicaba-SP: University of Sao Paulo; 2015

[10] Jin YS, Cate JHD. Metabolic engineering of yeast for lignocellulosic biofuel production. Current Opinion in Chemical Biology. 2017;**41**:99-106. DOI: 10.1016/j.cbpa.2017.10.025

[11] Kim SR, Skerker JM, Kong II, Kim H, Maurer MJ, Zhang GC, et al. Metabolic engineering of a haploid strain derived from a triploid industrial yeast for producing cellulosic ethanol. Metabolic Engineering. 2017;**40**:176-185. DOI: 10.1016/j.ymben.2017.02.006

[12] Estrela R, Cate JHD. Energy biotechnology in the CRISPR-Cas9 era. Current Opinion in Biotechnology. 2016;**38**:79-84. DOI: 10.1016/j. copbio.2016.01.005

[13] Jakociunas T, Jensen MK, Keasling JD. CRISPR/Cas9 adbances engineering of microbial cell factories. Metabolic Engineering. 2016;**34**:44-59. DOI: 10.1016/j.ymben.2015.12.003

**7**

Section 2

Animal Nutrition
