Renewable Biofuels: Bioconversion of Lignocellulosic Biomass by Microbial Community

116 Pages·2017·3.182 MB·English

by  Vandana Rana, Diwakar Rana (auth.)

SPRINGER BRIEFS IN
APPLIED SCIENCES AND TECHNOLOGY
Vandana Rana
Diwakar Rana
Renewable Biofuels
Bioconversion of
Lignocellulosic
Biomass by Microbial
Community
123
SpringerBriefs in Applied Sciences
and Technology
Series Editor
Janusz Kacprzyk
More information about this series at http://www.springer.com/series/8884
Vandana Rana • Diwakar Rana
Renewable Biofuels
Bioconversion of Lignocellulosic Biomass
by Microbial Community
Vandana Rana Diwakar Rana
Black & Veatch Corporation E. I. du Pont de Nemours and Company
Kansas City, Missouri, USA Overland Park, Kansas, USA
ISSN 2191-530X ISSN 2191-5318 (electronic)
SpringerBriefs in Applied Sciences and Technology
ISBN 978-3-319-47378-9 ISBN 978-3-319-47379-6 (eBook)
DOI 10.1007/978-3-319-47379-6
Library of Congress Control Number: 2016954434
© The Author(s) 2017
This work is subject to copyright. All rights are reserved by the Publisher, whether the whole or part of
the material is concerned, specifically the rights of translation, reprinting, reuse of illustrations, recitation,
broadcasting, reproduction on microfilms or in any other physical way, and transmission or information
storage and retrieval, electronic adaptation, computer software, or by similar or dissimilar methodology
now known or hereafter developed.
The use of general descriptive names, registered names, trademarks, service marks, etc. in this publication
does not imply, even in the absence of a specific statement, that such names are exempt from the relevant
protective laws and regulations and therefore free for general use.
The publisher, the authors and the editors are safe to assume that the advice and information in this book
are believed to be true and accurate at the date of publication. Neither the publisher nor the authors or the
editors give a warranty, express or implied, with respect to the material contained herein or for any errors
or omissions that may have been made.
Printed on acid-free paper
This Springer imprint is published by Springer Nature
The registered company is Springer International Publishing AG
The registered company address is: Gewerbestrasse 11, 6330 Cham, Switzerland
Abbreviations
ANOVA Analysis of variance
CCD Central composite design
CS-1 and LP-1 Solid loading, 50 g/L and enzyme loading, 5 FPU/mL
CS-2 and LP-2 Solid loading, 50 g/L and enzyme loading, 15 FPU/mL
CS-3 and LP-3 Solid loading, 100 g/L and enzyme loading, 5 FPU/mL
CS-4 and LP-4 Solid loading, 100 g/L and enzyme loading, 5 FPU/mL
EP Enzyme protein
FPU Filter paper unit
GY Glucose yield
HPLC High performance liquid chromatography
RSM Response surface methodology
SC Substrate concentration
SEM Scanning electron microscopy
WECS Wet exploded corn stover
WELP Wet exploded loblolly pine
WEx Wet explosion pretreatment
v
Contents
1 Background ............................................................................................... 1
1.1 Introduction ........................................................................................ 1
1.2 Lignin: A Major Barrier in Enzymatic Degradation .......................... 2
1.2.1 Overview of Lignin Structure and Function .......................... 2
1.2.2 Lignin Characterization ......................................................... 3
1.2.3 Pretreatment to Break the Lignin Recalcitrance .................... 5
References .............................................................................................. 15
2 Role of Microorganisms in Lignocellulosic Biodegradation ................. 19
2.1 Introduction ........................................................................................ 19
2.2 Lignocellulose Structure .................................................................... 22
2.2.1 Cellulose ................................................................................ 22
2.2.2 Hemicellulose ........................................................................ 23
2.2.3 Lignin ..................................................................................... 23
2.3 Enzymatic Hydrolysis of Lignocellulose ........................................... 25
2.3.1 Enzymatic Hydrolysis of Cellulose ....................................... 25
2.3.2 Enzymatic Hydrolysis of Hemicelluloses .............................. 26
2.3.3 Enzymatic Hydrolysis of Lignin ............................................ 27
2.4 Biodegradation by Microorganisms ................................................... 29
2.4.1 Cellulose Biodegradation ....................................................... 30
2.4.2 Hemicellulose Biodegradation ............................................... 32
2.4.3 Lignin Biodegradation ........................................................... 35
2.5 Lignocellulolytic Enzymes: Potential for Biorefinery ....................... 39
2.6 Conclusion ......................................................................................... 49
References ................................................................................................... 50
3 Enzyme Production from Trichoderma reesei
and Aspergillus Strain ............................................................................... 69
3.1 Introduction ........................................................................................ 69
3.2 Materials and Methods ....................................................................... 70
3.2.1 Wet Explosion-Alkaline Pretreatment of Corn Stover
for Cellulase Production from T. reesei RUT-C30 ................. 70
vii
viii Contents
3.2.2 Feedstock and Pretreatment (Wet Explosion) of Corn Stover
for β-Glucosidase Production from A. saccharolyticus ......... 71
3.2.3 Microorganisms and Inoculum Media ................................... 71
3.2.4 Fermentation Setup ................................................................ 72
3.2.5 Enzyme Characterization ....................................................... 73
3.2.6 Enzymatic Hydrolysis of Wet-Exploded Corn Stover
and Loblolly Pine ................................................................... 74
3.2.7 Analytical Methods ................................................................ 74
3.3 Results and Discussion ...................................................................... 74
3.3.1 Enzyme Production ................................................................ 74
3.3.2 Enzyme Characterization ....................................................... 76
3.3.3 Wet Explosion Pretreatment of Corn Stover and Loblolly
Pine for Enzymatic Hydrolysis Experiment .......................... 77
3.3.4 Evaluation of On-Site Produced Enzymes
by Enzymatic Hydrolysis Experiment ................................... 78
3.3.5 Comparison of On-Site and Commercial
Cellulolytic Enzyme Efficiency on Corn Stover .................... 78
3.3.6 Comparison of On-Site and Commercial Cellulolytic
Enzyme Efficiency on Loblolly Pine ..................................... 82
3.4 Conclusions ........................................................................................ 82
References ................................................................................................... 85
4 Use of Commercial Enzymes to Boost On-Site Enzyme Efficiency ...... 87
4.1 Introduction ........................................................................................ 87
4.2 Materials and Methods ....................................................................... 89
4.2.1 Enzymes ................................................................................. 89
4.2.2 Raw Material and Wet Explosion Pretreatment ..................... 89
4.2.3 Preparation of Inoculum ........................................................ 90
4.2.4 Enzyme Fermentation ............................................................ 91
4.2.5 Enzymes and Activities .......................................................... 91
4.2.6 Enzymatic Hydrolysis of Wet-Exploded Loblolly
Pine (WELP) .......................................................................... 92
4.2.7 Analytical Methods and Calculations .................................... 92
4.3 Results and Discussion ...................................................................... 93
4.3.1 Enzyme Activities .................................................................. 93
4.3.2 Raw Material and Wet Explosion Pretreatment
of Loblolly Pine ..................................................................... 93
4.3.3 Enzymatic Hydrolysis ............................................................ 94
4.4 Conclusion ......................................................................................... 101
References ................................................................................................... 102
5 Summary and Future Outlook ................................................................ 105
List of Figures
Fig. 1.1 Three major building blocks of monolignols of lignin ........................ 3
Fig. 1.2 Most common linkages within softwood lignin .................................. 4
Fig. 1.3 Schematic showing the effect of pretreatment effect
on lignocellulosic plant cell wall disruption ........................................ 5
Fig. 1.4 Pretreatment followed by enzymatic hydrolysis to release
the sugars from lignocellulosic biomass .............................................. 6
Fig. 1.5 Suggested reaction mechanism of the conversion
of H-units in lignin to S-units via in situ methoxylation
reactions during wet oxidation pretreatment; (a) five possible
resonant structures generated as a result of oxidation
via the action of oxygen on H-units, (b) demethoxylation
of polysaccharides followed by methoxylation of H-units .................. 10
Fig. 2.1 Molecular architecture of woody tissues and wood cell walls.
ML middle lamella, P primary wall, S1, S2, and S3 outer,
middle, and inner layers of secondary wall. Black parts
of middle lamella and cell walls show the presence of lignin
(T.K. Kirk: Marcus Wallenberg Foundation Symposia
Proceedings 2, p. 27 (1985)) ................................................................ 23
Fig. 2.2 Cellulase catalytic mechanism ............................................................. 25
Fig. 2.3 Xylan structure with different substituents and sites
of attack by xylanases (Beg et al. 2001) .............................................. 27
Fig. 2.4 Lignin biodegradation including enzymatic reactions
and oxygen activation (for explanation, see text).
Updated from Gutiérrez and Martínez ................................................. 29
Fig. 2.5 Simplified flow diagram of the enzymatic hydrolysis
using on-site enzyme from T. reesei .................................................... 41
ix
x List of Figures
Fig. 3.1 Dissolved oxygen saturation (%), temperature (°C), pH,
glucose concentration (gL−1), and protein content
(mg mL−1) during fermentation of T. reesei RUT-C30 ........................ 75
Fig. 3.2 Filter paper activity (FPU mL−1), carboxymethyl cellulose
activity, CMCase (U mL−1), β-glucosidase activity (U mL−1),
and xylanase activity (nkat mL−1) during fermentation
of T. reesei RUT-C30 ........................................................................... 76
Fig. 3.3 Monomeric sugar yield obtained from the hydrolysis
of wet-exploded corn stover with on-site enzymes
(T. reesei and A. saccharolyticus) and commercial enzymes
(Celluclast 1.5L and Novozym 188) at 5 and 10 % substrate
concentrations and 5 FPU and 10 CBU loadings ................................ 79
Fig. 3.4 Monomeric sugar yield obtained from the hydrolysis
of wet-exploded corn stover with on-site enzymes
(T. reesei and A. saccharolyticus) and commercial enzymes
(Celluclast 1.5L and Novozym 188) at 5 and 10 % substrate
concentrations and 15 FPU and 30 CBU loadings .............................. 80
Fig. 3.5 Monomeric sugar yield obtained from the hydrolysis
of wet-exploded loblolly pine with on-site enzymes
(T. reesei and A. saccharolyticus) and commercial enzymes
(Celluclast 1.5L and Novozym 188) at 5 and 10 % substrate
concentrations and 5 FPU and 10 CBU loadings ................................ 83
Fig. 3.6 Monomeric sugar yield obtained from the hydrolysis
of wet-exploded loblolly pine with on-site enzymes
(T. reesei and A. saccharolyticus) and commercial enzymes
(Celluclast 1.5L and Novozym 188) at 5 and 10 % substrate
concentrations and 15 FPU and 30 CBU ............................................. 84
Fig. 4.1 Sugars (g/L) released after 72 h of enzymatic hydrolysis
as an effect of in-house produced cellulase supplementation
with commercial cellulase, Cellic®Ctec2, from wet-exploded
loblolly pine ......................................................................................... 95
Fig. 4.2 Sugars (g/L) released after 72 h of enzymatic hydrolysis
as an effect of in-house produced cellulase supplementation
with commercial hemicellulase, Cellic®Htec2,
from wet-exploded loblolly pine ......................................................... 96
Fig. 4.3 Sugars (g/L) released after 72 h of enzymatic hydrolysis
as an effect of in-house produced cellulase supplementation
with commercial β-glucosidase, Novozym 188,
from wet- exploded loblolly pine ......................................................... 96
Fig. 4.4 Glucose, xylose, and mannose yields after 72 h of hydrolysis
for commercial cellulase, Cellic®Ctec2, supplementation
of constant in-house produced cellulase loadings
for wet-exploded loblolly pine ............................................................ 97
List of Figures xi
Fig. 4.5 Glucose, xylose, and mannose yields after 72 h of hydrolysis
for commercial hemicellulase, Cellic®Htec2, supplementation
of constant in-house produced cellulase loadings
for wet- exploded loblolly pine ............................................................ 97
Fig. 4.6 Glucose, xylose, and mannose yields after 72 h of hydrolysis
for commercial β-glucosidase, Novozym 188, supplementation
of constant in-house produced cellulase loadings
for wet-exploded loblolly pine ............................................................ 98