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BIOENERGY RESEARCH Evaluates challenges and sustainable solutions associated with various biofuel technologies Bioenergy Research offers an authoritative guide to recent developments in green bioenergy technologies that are currently available including: bioethanol, biobutanol, biomethanol, bio-oil, biohydrogen, biogas and biomethane. The authors provide in-depth analysis and discuss the commercial viability of the various technological advances in bioenergy. Comprehensive in scope, the book explores the environmental, practical and economic implications associated with a variety of bioenergy options. The book also considers the rollback of fossil fuels, the cost and their replacement as well as practical solutions for these issues. This important resource: * Presents up-to-date research and industrial developments for various bioenergy options * Offers comparative evaluation of bioenergy technologies for commercial feasibility * Reviews current challenges and sustainable solutions for a variety of biofuel technologies * Contains a review of existing strategies for bioenergy production Bioenergy Research is a valuable guide for academic researchers and industrial scientists working in the fields of biofuels and bioenergy, environmental science and technology, microbial technology, bioprocess engineering, and waste valorization.

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Table of Contents

Cover

Title Page

Copyright

List of Contributors

Foreword

Acknowledgments

Biofuels Production Technologies: Recent Advancement

1 Introduction

2 Bioethanol

3 Biodiesel

4 Biohydrogen

5 Biogas

6 Conclusion

Acknowledgement

References

1 Role of Enzymes in Biofuel Production

1.1 Introduction

1.2 Biofuel Classification

1.3 Enzymes Role in Biofuels

1.4 Enzymatic Reaction

1.5 Enzyme Recovery and Reuse

1.6 Enzyme Immobilization

1.7 Unique Techniques of Enzyme Immobilization

1.8 Application of Various Enzymes in Biofuel Production

1.9 Biofuel Production Process

1.10 Production of Biodiesel by Enzymatic Catalysis

1.11 Future Prospects

1.12 Conclusion

References

2 Microbial Technology for Biofuel Production

2.1 Introduction

2.2 Microbial Biofuel

2.3 Microbial Pathway for Biofuel Production

2.4 Algal Biofuel Production

2.5 Bioethanol

2.6 Biodiesel

2.7 Biohydrogen

2.8 Applications of Biofuel Production

2.9 Conclusion

References

3 Biohydrogen Production from Cellulosic Waste Biomass

3.1 Introduction

3.2 History of Hydrogen Fuel

3.3 Biohydrogen Fuel Cell

3.4 Cellulosic Biohydrogen Production from Waste Biomass

3.5 Conclusion

References

4 Strategies for Obtaining Biofuels Through the Fermentation of C5‐Raw Materials: Part 1

4.1 The Nature of Pentoses

4.2 Alcoholic Fermentation of C5

4.3 Lipid Biosynthesis from C5

4.4 Conclusion

References

5 Strategies for Obtaining Biofuels Through the Fermentation of C5‐Raw Materials: Part 2

5.1 Introduction

5.2 Ethanol Production Using C5‐Fermenter Strain

5.3 Microbial Lipid Production by C5‐Fermenter Strains for Biofuel Advances

5.4 Concluding Remarks

References

6 An Overview of Microalgal Carotenoids: Advances in the Production and Its Impact on Sustainable Development

6.1 Introduction

6.2 Diverse Category of Carotenoids

6.3 Microalgae Prospects for the Production of Carotenoids

6.4 Significance of Carotenoid in Human Health

6.5 Opportunities and Challenges in Carotenoid Production

6.6 Present Drifts and Future Prospects

6.7 Conclusion

References

7 Microbial Xylanases: A Helping Module for the Enzyme Biorefinery Platform

7.1 Introduction

7.2 Raw Material for Biorefinery

7.3 Structure of Lignocellulosic Plant Biomass

7.4 The Concept of Biorefinery

7.5 Role of Enzymes in Biorefinery

7.6 Enzyme Synergy: A Conceptual Strategy

7.7 Factors Affecting Biological Pretreatment

7.8 Advantages of Xylanases from Thermophilic Microorganisms in Biorefinery

7.9 The Products of Biorefinery

7.10 Molecular Aspects of Enzymes in Biorefinery

7.11 Conclusion

References

8 Microbial Cellulolytic‐Based Biofuel Production

8.1 Introduction

8.2 Biofuel Classifications

8.3 Bioprocessing of Bagasse for Bioethanol Production

8.4 Microbial Cellulase

8.5 Mode of Economical Production of Enzyme

8.6 Structure of Cellulase

8.7 Family Classification

8.8 Consortia‐Based Cellulase Production

8.9 Cellulase Production SSF Mode

8.10 Concluding Remarks

Declarations

Acknowledgment

References

9 Recent Developments of Bioethanol Production

9.1 Introduction

9.2 Emerging Techniques in Bioethanol Production

9.3 Advancement in Distillation and Waste‐Valorization Techniques

9.4 Green Extraction of Bioactive Products

9.5 Advancement in Bioethanol Production from Microalgae

9.6 Fermentation Technique Advances

9.7 Conclusion

References

10 Algal Biofuels – Types and Production Technologies

10.1 Introduction

10.2 Algal Biofuels

10.3 Production of Algal Biofuels

10.4 Types of Algal Biofuels

10.5 Advantages of Algal Biofuels

10.6 Limitations

10.7 Conclusion

References

11 Biomethane Production and Advancement

11.1 Introduction

11.2 Advancement Undergoing in the Process of Methane Production

11.3 Adsorption Methods

11.4 Separation by Membrane

11.5 Cryogenic Separation

11.6 Biological Technique for Purification of Biogas

References

12 Biodiesel Production and Advancement from Diatom Algae

12.1 Introduction

12.2 Diatom Algae as a Source of Lipids

12.3 Biodiesel Production from Diatoms

12.4 Innovative Approaches toward Enhancement in Biodiesel Production and Challenges

12.5 Advancements in Diatoms‐Based Biodiesel Production

12.6 Conclusion

Acknowledgments

References

13 Biobutanol Production and Advancement

13.1 Introduction

13.2 Biobutanol

13.3 ABE Process for Biobutanol Production

13.4 Biobutanol Production by ABE

13.5 Substrate Used in Biobutanol Production

13.6 Advancement in Pretreatment Method

13.7 Microbial Engineering for Production Enhancement

13.8 Conclusion

Acknowledgment

References

Index

End User License Agreement

List of Tables

f06

Table 1 Comparisons of different types of biofuel.

Chapter 1

Table 1.1 Benefits and drawbacks of biofuel.

Table 1.2 Biofuel classification with examples based on the sources.

Table 1.3 Applications of different enzymes in different industries.

Table 1.4 Unique methods for biodiesel production with their advantages and d...

Table 1.5 Biodiesel production with various lipases.

Table 1.6 Compounds present in diesel and biodiesel (Kulkarni et al. 2008).

Chapter 2

Table 2.1 Challenges in photolysis.

Table 2.2 Advantages and limitations of dark fermentation.

Table 2.3 Comparison of important biological hydrogen production processes.

Chapter 3

Table 3.1 Historical events in the development of the hydrogen fuel cell.

Table 3.2 Various cellulosic biomass produces different amount of hydrogen.

Chapter 4

Table 4.1 Occurrence of types of hemicellulose in cell walls of different gro...

Table 4.2 Fundamental constituents of hemicelluloses from different biomasses...

Table 4.3 End‐products and theoretical yields in microbial fermentative pathw...

Table 4.4 Fatty acid profile found in bio oil recovered from processes carrie...

Chapter 5

Table 5.1 Examples of microorganisms capable of converting xylose and or arab...

Table 5.2 Lipid production using different oleaginous species with xylose or ...

Chapter 6

Table 6.1 Different kinds of carotenoid produced by various type of microalga...

Table 6.2 Different methods of carotenogenesis induction in microalgae for ca...

Chapter 7

Table 7.1 Thermostable xylanases and their properties.

Table 7.2 Role of xylanases in the field of biorefinery.

Table 7.3 Genetically modified xylanases in the field of biorefinery.

Chapter 8

Table 8.1 Feedstock and ethanol production shown in (%).

Table 8.2 List of microbes genetically modified (Bischof et al. 2016).

Table 8.3 Biomass detail composition can be found in Zhang (2019).

Table 8.4 Cellulosome microbes.

Table 8.5 Mode of cellulase production (SSF SMF).

Chapter 9

Table 9.1 The generations of bioethanol.

Table 9.2 Advantages and disadvantages of green extraction methods.

Table 9.3 The publications for the application of the PFE method in bioethano...

Table 9.4 Bioethanol production from various food wastes.

Table 9.5 Bioethanol production from various solid wastes.

Chapter 10

Table 10.1 Microalgal growth types.

Table 10.2 Types of algae cultivation systems.

Table 10.3 Types of algal biofuels.

Table 10.4 Oil content of microalgae species.

Table 10.5 Carbohydrate content of microalgae species.

Chapter 12

Table 12.1 Different diatom species and their relative lipid content.

Table 12.2 Advantages and disadvantages of algal production methods.

Table 12.3 Different catalyst and their advantages and disadvantages.

Chapter 13

Table 13.1 Here is the detail about the various types of liquid biofuel avail...

Table 13.2 The table shows a short description how biofuels attained attentio...

Table 13.3 Many substrates are tested for the production of biobutanol. Here ...

List of Illustrations

f06

Figure 1 (a) Structure of lignocellulosic biomass and its biopolymers; cellu...

Figure 2 Schematic representation [adopted with permission from Wei et al. ...

Figure 3 Microalgae selection and biodiesel production.

Figure 4 Different process and route for biohydrogen production.

Figure 5 Main components and general process flow of biogas production [Tefe...

Chapter 1

Figure 1.1 Enzyme immobilization techniques.

Figure 1.2 Ethanol production by wet milling method.

Figure 1.3 Ethanol production by dry milling method.

Figure 1.4 Transesterification reaction for biodiesel production using lipas...

Figure 1.5 Comparative analysis of biodiesel production by enzymatic and alk...

Figure 1.6 Flow chart for biodiesel production.

Figure 1.7 Packed‐bed column method for biodiesel production.

Chapter 2

Figure 2.1 Biochemical pathway for biofuel production.

Figure 2.2 Techniques for Algal biofuel production.

Figure 2.3 Photosynthetic microbial fuel cell (MFC).

Figure 2.4 Advantages of bioethanol.

Figure 2.5 Process of transesterification.

Figure 2.6 Hydrogen production routes from fungi.

Figure 2.7 Applications of biofuel production.

Chapter 3

Figure 3.1 A representation of the significance of the hydrogen fuel. If hyd...

Figure 3.2 In the hydrogen fuel cell, hydrogen and oxygen are fed, and elect...

Figure 3.3 Structure of cellulose.

Figure 3.4 Free cellulose on hydrolysis produces glucose, which is enzymatic...

Figure 3.5 During dark fermentation, cellulose is converted into glucose thr...

Chapter 4

Figure 4.1 Monosaccharides and other chemical species found as components of...

Figure 4.2 Pathways for the conversion of D‐xylose to D‐xylulose‐5‐phosphate...

Figure 4.3 Conversion reactions of L‐arabinose to D‐xylulose‐5‐phosphate. (a...

Figure 4.4 Pentose‐phosphate pathways and its connection with the fermentati...

Figure 4.5 Embden‐Meyerhof‐Parnas pathway and alcoholic fermentation, integr...

Figure 4.6 Heterolactic fermentative pathway associated with pentose ferment...

Figure 4.7 Entner‐Doudoroff fermentative pathway and the artificial possibil...

Figure 4.8 Pentose assimilation and lipid biosynthesis in yeast and filament...

Chapter 6

Figure 6.1 Different types of carotenoid; (a) β‐carotene, (b) lutein, (c) as...

Figure 6.2 Systematic representation of carotenogenesis process of β‐caroten...

Figure 6.3 Shows how the oxidative stress induces the carotenogenesis in mic...

Figure 6.4 Systematic representation of overall process of β‐carotene produc...

Chapter 7

Figure 7.1 Brief overview of biorefinery process.

Figure 7.2 Structure of lignocellulosic plant biomass.

Figure 7.3 Role of enzymes in biological pretreatment and hydrolysis.

Figure 7.4 Synergistic action of enzymes.

Figure 7.5 Generation of butanol using agricultural residues.

Chapter 8

Figure 8.1 Biofuel generation.

Figure 8.2 Flow chart of separating sugar from bagasse.

Source:

From Dias et...

Figure 8.3 Common procedure to obtain the ethanol using cellulosic feedstock...

Figure 8.4 Diversity of enzymes required for lignocellulosic hydrolysis adap...

Figure 8.5 Different reaction by cellulase.

Figure 8.6 DOMAIN structure of CBH1, CBH2, and CBH3.

Chapter 9

Figure 9.1 (a) Schematic representation shows bioethanol production from sec...

Figure 9.2 Four generations of biofuel.

Figure 9.3 Various biological conversion process for bioethanol production f...

Figure 9.4 (a) SEM image of original ZSM‐5 zeolites, (b) SEM image of ZSM‐5 ...

Figure 9.5 Diagram exhibited the working principle of the MF‐NF‐DCMD system ...

Figure 9.6 Sustainable production of bio ethanol from agroindustrial waste....

Figure 9.7 Flow chart showing bioethanol production from solid waste.

Chapter 10

Figure 10.1 Types of mariculture methods used in seaweed cultivation. a. Lon...

Figure 10.2 Types of microalgal culture systems.

Figure 10.3 Types of open pond systems for microalgae cultivation. a. Unstir...

Figure 10.4 Types of photobioreactors. a. Flat Plate reactor b. Bubble Colum...

Figure 10.5 Overview of microalgae harvesting methods.

Figure 10.6 Transesterification of triglyceride to biodiesel.

Figure 10.7 Stages of anaerobic digestion in the process of biogas productio...

Chapter 12

Figure 12.1 Schematic representation of various generation of biofuel.

Figure 12.2 Flow chart showing biodiesel production from diatoms.

Chapter 13

Figure 13.1 Schematic representation of the conversion of glucose into solve...

Guide

Cover

Table of Contents

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Bioenergy Research

Evaluating Strategies for Commercialization and Sustainability

Edited byNeha SrivastavaIIT (BHU)Chemical Engineering & Technology221005 VaranasiIndia

Manish SrivastavaIIT (BHU)Chemical Engineering & Technology221005 VaranasiIndia

 

 

 

 

 

 

 

 

 

 

This edition first published 2021

© 2021 by John Wiley & Sons Ltd

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Library of Congress Cataloging‐in‐Publication Data

Name: Srivastava, Neha, 1981‐ editor. | Srivastava, Manish, editor.

Title: Bioenergy research : evaluating strategies for commercialization and

 sustainability / edited by Neha Srivastava, IIT (BHU) Chemical

 Engineering & Technology, Manish Srivastava, IIT (BHU) Chemical

 Engineering & Technology.

Description: First edition. | Hoboken, NJ : Wiley, 2021. | Includes

 bibliographical references and index.

Identifiers: LCCN 2020051125 (print) | LCCN 2020051126 (ebook) | ISBN

 9781119772095 (cloth) | ISBN 9781119772101 (adobe pdf) | ISBN

 9781119772118 (epub)

Subjects: LCSH: Biomass energy. | Renewable energy sources.

Classification: LCC TP339 .B533 2021 (print) | LCC TP339 (ebook) | DDC

 662/.88–dc23

LC record available at https://lccn.loc.gov/2020051125

LC ebook record available at https://lccn.loc.gov/2020051126

Cover Design: Wiley

Cover Image: © esemelwe/iStock/Getty Images

List of Contributors

Komal Agrawal

Bioprocess and Bioenergy Laboratory

Department of Microbiology

Central University of Rajasthan

Ajmer

Rajasthan

India

Mayara C.S. Barcelos

Laboratory of Food Biotechnology

Institute of Science and Technology

UFVJM Diamantina

MG

Brazil

Nisha Bhardwaj

Bioprocess and Bioenergy Laboratory

Department of Microbiology

Central University of Rajasthan

Ajmer Rajasthan

India

S.M. Bhatt

Agriculture Department

ACET Amritsar

Amritsar Group of Colleges

Punjab

India

Sashita Bindu Ekka

Department of Environmental Science

Indira Gandhi National Tribal University

Amarkantak

MP

India

N.B. Chandrasekhar

Research and Development Center Department of Biotechnology

Shridevi Institute of Engineering and Technology

Tumakuru

India

S. Dheiver

Institute of Computing

Federal University of Alagoas (UFAL)

Maceio

Brazil

Stephen Edward Chattree

Galgotias University

Greater Noida

UP

India

S K. Godlaveeti

Centre for Nanoscience and Nanotechnology

Sathayabama Institute of Science and Technology

Chennai

India

Rahul Kumar Goswami

Bioprocess and Bioenergy Laboratory

Department of Microbiology

Central University of Rajasthan

Ajmer

Rajasthan

India

Quyet Van Le

Faculty of Natural Sciences

Duy Tan University

Danang

Vietnam

P K. Mishra

Department of Chemical Engineering and Technology

Indian Institute of Technology (BHU)

Varanasi

India

Gustavo Molina

Laboratory of Food Biotechnology

Institute of Science and Technology

UFVJM Diamantina

MG

Brazil

Thi Hong Chuong Nguyen

Institute of Research and Development

Duy Tan University

Da Nang

Vietnam

and

Faculty of Natural Sciences

Duy Tan University

Danang

Vietnam

Lílian A. Pantoja

Institute of Science and Technology

Federal University of Jequitinhonha and Mucuri Valleys UFVJM Diamantina

MG

Brazil

Veena Paul

Department of Dairy Science and Food Technology

Institute of Agricultural Sciences

Banaras Hindu University

Varanasi

UP

India

Enosh Phillips

Department of Biotechnology

St. Aloysius College (Autonomous)

Jabalpur

Madhya Pradesh

India

Ram Prasad

Department of Botany

Mahatma Gandhi Central University

Motihari

Bihar

India

Spriha Raven

Diatom Research Laboratory

Amity Institute of Biotechnology

Amity University

Noida

UP

India

Lipi Rina

Department of Agronomy

SHUATS

Prayagraj

UP

India

Shivani Smita Sadanand

Department of Agriculture

SHUATS

Prayagraj

UP

India

Arla Sai Kumar

Department of Materials Science and Nanotechnology

Yogi Vemana University

Kadapa

Andhra Pradesh

India

Sana Siva Sankar

School of Chemical Engineering and Technology

North University of China

Taiyuan

China

Alexandre S. Santos

Department of Basic Science

Federal University of Jequitinhonha and Mucuri Valleys UFVJM Diamantina

MG

Brazil

Alexandre Soares dos Santos

Department of Basic Science

Federal University of Jequitinhonha and Mucuri Valleys ‐ UFVJM – Diamantina

MG

Brazil

Sreedevi Sarsan

Department of Microbiology

St. Pious X Degree and PG College

Hyderabad

India

Abhishek Saxena

Diatom Research Laboratory

Amity Institute of Biotechnology

Amity University

Noida

Uttar Pradesh

India

Akshay Shrivastav

Department of Chemical Engineering

Madan Mohan Malaviya University of Technology

Gorakhpur

India

Rajeev Singh

Department of Chemical Engineering and Technology

Indian Institute of Technology (BHU)

Varanasi

India

Neha Srivastava

Department of Chemical Engineering and Technology

Indian Institute of Technology (BHU)

Varanasi

India

K.R. Srivastava

Department of Chemical Engineering and Technology

Indian Institute of Technology (BHU)

Varanasi

India

Manish Srivastava

Department of Chemical Engineering and Technology

Indian Institute of Technology (BHU)

Varanasi

India

Archana Tiwari

Diatom Research Laboratory

Amity Institute of Biotechnology

Amity University

Noida

UP

India

Abhishek Dutt Tripathi

Department of Dairy Science and Food Technology

Institute of Agricultural Sciences

Banaras Hindu University

Varanasi

UP

India

K. Vindhya Vasini Roy

Department of Microbiology

St. Pious X Degree and PG College

Hyderabad

India

Pradeep Verma

Bioprocess and Bioenergy Laboratory

Department of Microbiology

Central University of Rajasthan

Ajmer

Rajasthan

India

Kele A. C. Vespermann

Laboratory of Food Biotechnology

Institute of Science and Technology

UFVJM Diamantina

MG

Brazil

Ashok Kumar Yadav

Food Processing and Management

DDU Kaushal Kendra

Rajiv Gandhi South Campus

Banaras Hindu University

Barkachha

Mirzapur

UP

India

Foreword

To discontinue the chain of harmful impact of environmental pollution, fossil fuel issues must be resolved in a sustainable manner. Production of renewable energy from renewable resources is one of the most effective alternatives toward the replacement of fossil fuels. For decades, research on renewable energy production has been studied for its commercialization purpose in an environment‐friendly manner; the various downsides keep it stuck, which hampers its commercial implementation effectively and uniformly. To date, many renewable energy options have been explored and analyzed for their commercial sustainability. Bioenergy production is the most sustainable alternative to replace fossil fuels, and rigorous research has been done on bioenergy topics as the most sustainable solution. Although the number of bioenergy options are available and detailed research has been performed, the sustainable bioenergy options are still far away from commercialization and practical viability. There is an urgent need to summarize and critically evaluate the available bioenergy options for the practical implementation for the replacement of fossil fuels.

Publication of Bioenergy Research: Evaluating Strategies for Commercialization and Sustainability is a useful approach in this area. I am writing this message with joy as a working researcher in this area. This book holds 13 chapters, presented as an evaluation of different bioenergy options with complete details and up‐to‐date research as well as development. The book expands on different bioenergy options, such as biohydrogen, bioethanol, and biomass to fuel production and algal biofuels, and so on. Additionally, the book expands upon the feasibility of these biofuels on the basis of their properties, advantages, drawbacks, research updates, and existing hurdles still in the way of their hopeful implementation. In my introduction, I talk about how this book will serve as a preliminary introduction to anyone working in the relevant areas, both academic and research institutions as well as industrial.

I appreciate the efforts of Dr. Neha Srivastava and Dr. Manish Srivastava for Bioenergy Research: Evaluating Strategies for Commercialization and Sustainability. The efforts taken to build this book will surely cover the gap between the academic research benches to a practical industrial‐scale study. I congratulate the editors for their hard work in bringing a final shape to this book.

Dr Vijai Kumar Gupta

Center for Safe and Improved Food &

Biorefining and Advanced Biomaterials Research Center

Scotland's Rural College (SRUC), UK

Acknowledgments

The editors are thankful to all the academicians and scientists whose contributions have enriched this volume. We also express our deep sense of gratitude to our parents whose blessings have always prompted us to pursue academic activities deeply. It is quite possible that in a work of this nature, some mistakes might have crept in text inadvertently and for these we owe undiluted responsibility. We are grateful to all authors for their contribution to present book. We are also thankful to Wiley for giving this opportunity to editors and Department of Chemical Engineering & Technology, IIT (BHU) Varanasi, U.P., India for all technical support. We thank them from the core of our heart.

Biofuels Production Technologies: Recent Advancement

Neha Srivastava and Manish Srivastava

Department of Chemical Engineering and Technology, Indian Institute of Technology (BHU), Varanasi, 221005, India

1 Introduction

Fossil fuels are the hydrocarbon, coal, or natural gas that is produced from the dead remains of living organisms. The combustion of these fuels by humans causes the emission of carbon dioxide, sulfur dioxide, nitrous oxide, methane, etc., which causes harmful effects and contributes to global warming and pollutes the environment (Basu et al. 2020). The excessive utilization of fossil fuels has also led to its shortage. Fossil fuels are essential to fulfill global energy needs, accounting for more than 80% of the world's primary energy consumption (Karnauskas et al. 2020).

In order to reduce fossil fuel usage, other renewable and sustainable sources of energy, like solar power, wind energy, wave energy, and thermal energy from the earth's crust, as well as biofuels may be taken into consideration (Jia et al. 2018). Biofuels are one of the best green alternatives, which have the potential to replace fossil fuels because they are ecofriendly, easily available, completely combustible, and nontoxic. A biofuel is defined as a source of energy or a fuel, which is obtained using a biological carbon fixation process. It is the most productive form of renewable energy source, as it is generated from renewable sources such as biomass (Milano et al. 2016). It is widely acknowledged that biofuels combustion does not contribute to greenhouse effect. Biofuel not only reduces the dependency of the world toward oil but also decreases the emission of harmful gases; moreover, its production can also provide new employment and income options in rural and urban areas. Low emission, nontoxic, environmentally friendly, and safer are some of the green and unique characteristics of biofuels, which make them potential candidates for their effective utilization (Kumarappan and Joshi 2011). Further, the biofuels are categorized as first‐generation, second‐generation, third‐generation, and fourth‐generation biofuel based on feedstocks from which they are produced (Bhatia et al. 2017).

Biofuel production from the first generation requires food crops that are commonly grown on arable lands as a substrate. This process utilizes the complete food crops only for producing fuel and not anything else. The process used in this generation was yeast fermentation or transesterification for production of ethanol or biodiesel (Rodionova et al. 2017). The second generation of biofuel production requires lignocellulosic biomass, human waste, woody chips, and agricultural by‐products as a substrate. The benefit of this process is that it utilizes any kind of biomass for the production of biofuel, but unlike the other processes, there are some difficulties in extraction of fuels. This process requires some extra steps, such as physical, chemical pretreatment just to obtain proper yield. The third generation of biofuel production utilizes oil‐rich algae as a substrate for production of alcohol. The types of biofuel production in the third generation varies because of the two different characteristics of microbes. First, the oil produced by algae can be refined to biodiesel or some content of gasoline, whereas in the second, genetic manipulation leads to production of ethanol, butanol, gasoline, and even biodiesel (Leong et al. 2018). In the fourth generation of biofuel, the destruction of biomass is not required and biofuels are produced from nonarable land. This category includes biofuels, such as photobiological solar fuels and electrofuels (Abdullah et al. 2019). There are different types of biofuels, such as bioethanol, methanol, biodiesel, biogas, bio‐butanol, and bio‐hydrogen, which exist in nature. Different types of biofuels can be produced by varying fermentation techniques and microorganisms. This variation and comparisons between the different types of biofuel are described in Table 1. There are currently two main types of biofuels used: ethanol and biodiesel.

Table 1 Comparisons of different types of biofuel.

Bioethanol

Biodiesel

Biohydrogen

Biogas

Substrate

Sugarcane, corn, and sugar beat

Vegetable oils

Food waste

Municipal solid waste

Process involved

Pretreatment Hydrolysis Fermentation Product upgradation

Pretreatment Hydrolysis Fermentation Lipid extraction Transesterification

Pretreatment Hydrolysis Fermentation

Pretreatment Hydrolysis Acidogenesis Acetogenesis Methanogenesis

Pre‐treatment preferred

Steam explosion

Biological pre‐treatment

Heating of inocula

Ultrasonification

Fermentation preferred

Solid state fermentation

Solid state fermentation

Dark fermentation

Anaerobic, fermentation

Microbes preferred

Bacteria and yeast

(Saccharomyces cerevisiae

and

Zymomonas mobilis)

Fungi (white rot fungi and brown rot fungi)

Bacteria (Lactobacillus,

Thermoanaerobaterium

, Clostridium)

Bacteria (Methanogens and acidogens)

Current research

Downstream process and separation of biomass after pre‐treatment

Cost‐effective microalgae and biophotoreactor

Fermentation condition and catalyst

Bioreactors, process of anaerobic digestion and application of high pressure

2 Bioethanol

The process of ethanol production is classified into four different categories, (i) pretreatment, (ii) hydrolysis, (iii) fermentation, and (iv) product upgradation. Pretreatments can be done by physical means (microwave radiation, pyrolysis, sonication, and spry drying), chemical means (hydrolysis, oxidation, ozonation, and alkaline pretreatment), and biological means (bacterial and enzymatic treatment). In the process of ethanol production, steam explosion is mostly used a pretreatment process at the commercial level (Trinh et al. 2019). Pretreatment of lignocellulosic biomass helps the exposure of cellulose and hemicelluloses and removes lignin as it ruptures the complete structure (Figure 1) (Karuppiah and Azariah 2019). After the pretreatment process, enzymatic or chemical hydrolysis takes place, which converts cellulose and hemicelluloses hexose, pentose, and glucose as a simple fermentable carbohydrate. These simple carbohydrates get fermented by the help of yeast, fungi, or bacteria using two different mechanisms of fermentation, (i) solid state fermentation and (ii) submerged fermentation (Brexó and Sant'Ana 2017). These C‐5 and C‐6 carbohydrates can be fermented to ethanol by the help of yeast and bacteria (Saccharomyces cerevisiae and Zymomonas mobilis are the most common species used at industrial level currently) (Gallagher et al. 2018; Nakamura and Shima 2018; Karuppiah and Azariah 2019) (Figure 2). At the end, the produced bioethanol has to go through a dehydration and distillation process for producing ethanol with 99.9% purity. Earlier hydrolysis and fermentation are carried out in two different chambers; nevertheless, the process can be carried out in the same chamber. Using these techniques, fermentation and saccharification, direct conversion, prehydrolysis, and simultaneous fermentation and saccharification, as well as simultaneous saccharification and cofermentation can be occurred simultaneously (Zabed et al. 2017; Chen and Xiaoguo 2016). In the process of ethanol production, the solid state fermentation technique is mostly used and this technique performs two different steps in the same chamber.

The application of lignocellulosic biomass, such as sugarcane, corn, and kitchen waste as a substrate for bioethanol production in currently trending across the world. Microbes, such as Codium tomentosum, Myceliophthora thermophila, S. cerevisiae, and moringaolefira are currently used at different levels to produce and test bioethanol (Konur 2020). Countries like the UK and the USA are working on the production of bioethanol using marine yeast and sea water as a media. In 2019, the United States produced the maximum of ethanol, which is around 15.8 billion gallons, while Brazil was in second position with 8.6 billion gallons. On this list, India is at the fifth position with 530 million gallons in 2019 (Aparicio et al. 2020). The most common substrate for ethanol production is starch released from sugar cane, corn, and sugar beets. The United States is the largest producer of corn in the world, which produced about 345.89 million metric tons in 2019. Therefore, the United States utilizes corn as a feedstock for ethanol production because of its abundance and low cost, whereas Brazil is the largest producer of sugar cane in the world and utilizes it as a substrate because of its easily availability and low cost (Mayumi 2020). Bioethanol productions are still not being used at commercial level in many countries and the problem may be due to their feed and food purposes. One of the major disadvantages with the production of bioethanol in developing countries is a lack of availability of feed stocks for production. As the production of bioethanol continues to grow, it is the compulsion of producing bioethanol using renewable substrates which are still not found. Currently, the downstream process in ethanol production and separation of biomass constituents after pretreatment are focused, and several points of research are under process for developing an economical procedure (Mohanty and Swain 2019).

Figure 1 (a) Structure of lignocellulosic biomass and its biopolymers; cellulose, hemicellulose, and lignin [Hernández‐Beltrán et al. (2019), CCBY 4.0]. (b) Pretreatment methods to increase the bioavailability of lignocellulosic biomass. [Hernández‐Beltrán et al. (2019), CCBY 4.0].

Figure 2

Schematic representation [adopted with permission from Wei et al. (2015)]. (a) and metabolic pathway of fermentation (b) for producing ethanol.

Source: El‐Dalatony et al. (2017). Licensed under CC BY 4.0.

3 Biodiesel

Biodiesel can be produced using substrate like oils from different seeds (rapeseeds, soyabeans, palm), animal waste (animal fats, fish oil, poultry oils), as well as other sources, such as almond, barley, and lignocellulosic biomasses (Chen et al. 2018). The process of biodiesel production starts with pretreatment process in which most favorable is biological techniques using different microorganisms (white rot fungi such as Irpexlacteus, P. floridensis, Ceriporiopsis subvermispora, Punctularia spp., Phlebia brevispora, brown rot fungi such as Meruliporia incrassate, Serpula lacrymans, Laetoporus sulphurous, Coniophora puteana) (Rudakiya and Gupte 2017; Hegnar et al. 2019; Wang et al. 2017). After the process of pretreatment, th e process of enzymatic hydrolysis starts at a mild climatic condition (pH 4.5–5.0 and temperature 40–50 °C). Fermentation techniques depend upon the state of substrate, whether it is liquid (submerged fermentation) or solid (solid state fermentation), whereas solid state technique is mostly preferred in biodiesel production. Biodiesel can be produced using microorganisms which have the ability to accumulate concentration of lipids of more than 20% (Sohedein et al. 2020). These microbes can be bacteria (Arthrobacter sp., Rhodococcus opacus, Acinetobacter calcoaceticus), yeasts (Cryptococcus albidus, Candida curvata, Rhodotorula glutinis), microalgae (Cylindrotheca sp, Schizochytrium sp, Botryococcus braunii), and fungi (Mortierella isabellina, Aspergillus oryzae, Mortierella vinacea) (Bardhan et al. 2020). The produced lipids can be extracted using process like organic solvent, microwave, bead beatings, etc., and the extracted lipids then have go through a direct transesterification process for the production of biodiesel.

Biodiesel is generally used in normal diesel engines alone or as a blend with petrodiesel. Different countries utilize different feedstock as per their availability; for example, different parts of Europe utilize sunflower and rapeseeds as a substrate, while soybean is commonly used in the United States, and canola oil is used in Canada, and in tropical countries, palm oils are used (Yesilyurt et al. 2020). Application of these food oils have raised an issue related to food versus fuel. To overcome this problem, production of biodiesels for different second‐ (pongamia, jatropha, karanja) and third‐ (microalgae and other microorganisms) generation feedstocks are currently trending with lots of research, both at the lab scale as well as at the industrial scale (Eiben et al. 2020). In 2019, the United States produced the maximum amount of biodiesel, which is about 6.5 billion liters, and Brazil is a little behind the United States with annual production of 5.9 billion liter of biodiesel in 2019. India produced about 2.5 billion liters of biodiesel in year 2018–2019. United States is the largest producer of soybeans in the world and produced about 120.52 million metric tons in 2018–2019 (Bognar et al. 2020). Therefore, the United States utilizes soybeans as a feedstock for biodiesel production because of its abundance and low cost. Brazil overtook the United States in soybean production in 2019–2020 with 124 million metric tons (Vunnava and Singh 2020). Currently, different research is going on for the preparation of cost‐effective microalgae and developing photobioreactors, which will increase the production of biodiesel (Atabani et al. 2017). Some important factors of using microalgae are high production, low cost, as well as easy availability as described in Figure  3, whereas some of the drawbacks in the process are low oil content, low concentration of biomass in the culture, and the small size of microalgae which increases the cost of harvest (Duong et al. 2012).

4 Biohydrogen

There are numerous types of pretreatments methods, such as microwave, heating, etc. However, the most important pretreatment method which is widely adopted by researchers is the heating of inocula for the production of biohydrogen (Prabakar et al. 2018). After the pretreatment process of biomass, enzymes can penetrate easily to cell walls and convert cellulose and hemicellulose to sugars that can be fermented easily via enzymatic hydrolysis (Algapani et al. 2016). Further, biohydrogen can be produced by different methods such as photo‐fermentation, indirect biophotolysis, dark fermentation, and direct biophotolysis (Figure 4) (Chandrasekhar et al. 2015). Among these, dark fermentative biohydrogen production has been reported to be a potential economic and effective method due to the high yield and efficiency using a variety of substrate (cellulose, sugary waste water, sugarcane juice, corn pulp, food waste). Microbes (especially bacteria), such as Lactobacillus, Thermoanaerobaterium, Clostridium, Rhodopseudomonas, Citrobacter, and Enterobacter are mainly used as fermentative microorganisms in the biohydrogen production process (Kumar et al. 2019). There are three different pathways of conversion of organic waste in biohydrogen using dark fermentation, favored by the thermodynamics process. These pathways are as follows: (i) acetic acid from hexose, (ii) butyric acid from hexose, and (iii) ethanol from acetate, and among these pathways, the acetate ethanol pathway has been found to be more stable as compared to the others. The microbes grow on biodegradable wastes, having a high quantity of sugar in order to produce pyruvate through glycolysis techniques, which get further oxidized into acetyl CoA through the reduction of ferredoxin while producing adenosine di‐phosphate (ATP) and acetate. Finally, ferredoxin is oxidized into hydrogen with the help of hydrogenase enzymes. This enzyme is responsible for the production of biohydrogen using dark fermentation (Liu et al. 2017).

Figure 3 Microalgae selection and biodiesel production.

Source: Duong et al. (2012). Licensed under CC BY 3.0.

Figure 4 Different process and route for biohydrogen production.

Source: Chandrasekhar et al. (2015). Licensed under CC BY 4.0.

In recent years, the production of biohydrogen using organic waste as a substrate is mostly found in developed countries like the USA, the UK, and Germany. In these countries, the application of two‐stage bioreactors for producing both hydrogen as well as methane are focused on. Research is ongoing on the utilization of food waste as a substrate for producing hydrogen (Hassan et al. 2019). As per the Food and Agriculture Organization(FAO), about one third of the world foods are wasted, which is approximately1.3 billion tons per year. According to the study performed by the Economist Intelligence Unit, food sustainability index (FSI), and as per the FSI 2017 report, Australia produced the highest quantity of food waste per capita (361 kg), whereas other countries, such as Sweden (200 kg), the USA (287 kg), Russia (56 kg), and China (44 kg) also produce a high amount of waste. Moreover, India also produces about 51 kg of kitchen waste per capita (Boliko 2019). Improvements as well as advancements are made, such as variation in different fermentation conditions like temperature and pH and concentration of substrate (carbon to nitrogen ratio), plays a significant role to increase the final yield. Apart from all these advancements and improvements, there are two major problems that act as a barrier in the path of biohydrogen production for being used at commercial scale. These problems are low yield and have a high operation cost, which acts as a serious issue in different countries (Ishangulyyev et al. 2019). Some latest advancements, such as application of nanoparticles, metal oxides and ions, and immobilization of microbes, are appealing in their prospects to achieve high yields as well as a profitable efficiency of the process (Yun et al. 2018).

5 Biogas

Biogas is a type of biofuel produced by anaerobic digestion of organic waste, such as biomass, cow dung, agricultural residue, green waste, sugar cane, and cassava, etc. The production of biogas is classified into different steps which occurs in an anaerobic reactor (Plugge 2017). These steps are as follows: pretreatment, hydrolysis, acidogenesis, acetogenesis, and methanogenesis (Campanari et al. 2017). There are lots of pretreatment processes which can be used in the process of biogas production, such as milling, preheating, steam explosion, and liquid hot water, etc. However, the most important pretreatment method which was widely adopted at the industrial level is the preheating by ultrasonification of substrate (Deepanraj et al. 2017). Hydrolysis is the primary step in the process of biogas production. This step is achieved by breaking down and solubilizing organic complex compounds to simple soluble compounds. Enzymatic hydrolysis is one of the most preferred processes which accelerates the process through oxidation of organic matter (Tongco et al. 2020). After the hydrolysis process, the substrate is available for transportation of cells and bacteria which will ferment the substrate during acidogenesis. In this process, the produced organic acids get transformed into hydrogen, carbon dioxide, acetic acid, and acid derivatives. In this process, the main role is being performed by microorganisms which are classified into two different groups based on the product they produce (Fernandez 2018). The two main groups of these microbes are the methane‐producing bacteria (methanogens –Methanoculleus, Methanosarcinales, Methanobacteriales) and acid‐producing bacterial (acidogens – Moorella thermoacetica, Clostridium formiaceticum, Acetobacter woodii, Clostridium termo autotrophicum). Methanogensis is one of the critical steps in the process of the acidogenesis because about 70% of methane used in anaerobic digestion is produced in this step only, whereas during acetogenesis ethanol, volatile fatty acids (VFAs) with more than two carbons get converted by acetate‐forming bacteria into carbon dioxide and hydrogen (main product) and acetate (Mulat et al. 2016). In this step, only methanogens convert hydrogen (oxidizing) and carbon dioxide (reducing) to methane, as well as acetolactic methanogens which convert acetate to methane. Therefore, the produced biogas contains 1–5% other gases, including hydrogen, carbon dioxide 35–40%, and methane 55–60%.

Recently, lots of work continues in the development of bioreactors, the separation of anaerobic digestion into two different steps, and the high‐pressure digestion which can increase the production of biogas. Developing countries have higher amounts of municipal solid waste, and about 74% of municipal solid waste generated in Indonesia, 75% generated in Bangladesh, 76.4% generated in Sri Lanka, 42% generated in India, and 52.6% generated in China is organic. China is the world's largest producer of municipal solid waste by producing 215 million tons per year and was expected to produce 480 million tons in 2030 (Cudjoe et al. 2020). This waste includes solid waste, such as papers, woods, meats, plastics, etc. China utilizes it as a substrate for biogas production. Therefore, China was the world's largest producer of biogas in 2014 with the total production of about 15 billion cubic meters. There is lots of ongoing research into the various upgrading and cleaning techniques in order to improve the quality of biogas (Xue et al. 2020). These improvements are classified on its behavior into two different groups – physiochemical and biological technologies. This technology includes cryogenic separation, hydrated separating technique, enriching membranes, anaerobic, high pressure, and multistage digestion (Theuerl et al. 2019; Passos et al. 2020). Still, there are lots of problems which is why biogas production cannot be implemented on a large scale in different countries. These problems are infrastructural challenges, due to the unavailability of feedstocks, improper segregation, and high installation cost. In spite of all these issues, biogas is one of the most important sources of energy, especially in rural areas, and it is also utilized as an alternative and renewable source of energy at a small scale whose flow sheet is explained in Figure 5 (Teferra and Wubu, 2018).

Figure 5 Main components and general process flow of biogas production [Teferra and Wubu (2018), CCBY 3.0].

6 Conclusion

In the present world, bioethanol and biodiesel are mostly produced and in demand as compared to other types of biofuel that exists in nature. Different developed countries, such as the USA, the UK, and Brazil, are the top producers of biodiesel and bioethanol. In 2019 the USA produced about 15.8 billion gallons of bioethanol and about 6.5 billion liters of biodiesel. Similar to this, Brazil produces about 5.9 billion liters of biodiesel and 8.6 billion gallons of bioethanol in the same working year. Lots of countries produce bioethanol because it can be produced by using large variety of carbohydrates as a substrate. It is easily possible to convert waste like woods, straws, and even daily use waste to bioethanol. Along with these, one of the most important reasons for producing bioethanol at this large of scale is that it can be used as a substituent or additive toward petrol. As fossil fuel decreases day by day, it is a major focus for every country to work on alternatives. Biodiesel is also an alternative of very important fossil fuel which can be easily used in any kind of diesel engines without even modifying it. Biodiesel is even better than diesel in terms of flashpoint, aromatic content, and using blend biodiesel compared to normal diesel gives even better fuel economy.

The basic difference between ethanol and biodiesel are that one is fuel and the other one is oil. Ethanol is an alcohol created through fermentation and can be used as a substitute or along with gasoline, whereas biodiesel is generated by extracting naturally occurring oils from differed plants and seeds by the process known as transesterification. The production of biofuel is focused on not only fulfilling the requirements of energy production at the decentralized level but also for fulfilling the requirements of transport. This generates interest from regional groups as well as that it involves the land of regional communities. This creates incentives for the local communities, especially if community lands are involved.

Acknowledgement

Author M.S. acknowledges the Science and Engineering Research Board for SERB Research Scientist award [SB/SRS/2018‐19/48/PS] and also to DST for DST INSPIRE Faculty award [IFA‐13‐MS‐02].

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1Role of Enzymes in Biofuel Production

Ashok Kumar Yadav1, Surabhi Pandey2, Abhishek Dutt Tripathi3, and Veena Paul2

1Food Processing and Management, DDU Kaushal Kendra, Rajiv Gandhi South Campus, Banaras Hindu University, Barkachha, Mirzapur, UP, India

2Department of Dairy Science and Food Technology, Institute of Agricultural Sciences, Banaras Hindu University, Varanasi, UP, India

1.1 Introduction

The International Energy Agency (2006) released a factsheet based on a survey stating that approximately 80.3% of fossil fuels utilized as a principal energy source, and 57.7% employed for transport purposes. The demand for energy will grow by about 37% by 2040 due to the significant change in the global energy system (International Energy Agency 2006