Technological Trajectories in Enzymatic Hydrolisis
José Maria Ferreira Jardim da Silveira & Thays MurakamiCenter of Agriculture and Environmental Research (NEA); Center of Interdisciplinary Research on Energy (NIPE/BE-BASIC & Institute of Economics (IE), University of Campinas (UNICAMP/), Brazil/
Paper prepared for presentation at the 19th ICABR ConferenceIMPACT OF THE BIOECONOMY ON AGRICULTURAL SUSTAINABILITY, THE ENVIRONMENT AND HUMAN HEALTH Ravello, June 17-19, 2015Copyright 2015 by author(s). All rights reserved. Readers may make verbatim copies of this document for non-commercial purposes by any means, provided that this copyright notice appears on all such copies.
Motivation: description of the technologyEnzimatic Hydrolisis in the context of Second Generation
Motivation of the paper: what is the role of Enzymatic Hydrolisis (EH) ?
Scenarios of Brazil:
link between products, technological routes and biomass;
First generation still has an expected technological evolution;
The concept of modularity/specialization shows up;
The possibilities of co-evolution of different trajectories: first generation and second generation;
Simultaneous fermentation 5C with 6C as a focusing device, but separate 5C fermentation (Xylose and others) is still another option for tech evolution: microorganisms, enzymes and biomass.
Short term Medium term Long term
2G Perspectives
1 Medium and long term scenarios include a small 1G plant processing energy cane juice
2 Including straw
Increase on 2G yields and recoveries
2G1 All year: sugarcane bagasse+ straw
All year: energy cane bagasse2
All year: energy cane bagasse2
1G2G Season: sugarcane bagasse+ straw
Season: sugarcane bagasse+ straw
Off-season: energy cane bagasse2
Season: sugarcane bagasse+ straw + energy
cane bagasse2
Off-season: energy cane bagasse2
Source: Junqueira, 2015 CTBE/CNPEM
2G Flowsheet
5
Short term and “A” scenarios– Separated C5 fermentation
C5 fermentation - Partial cell recycling is considered.
2G Flowsheet
6
C5/C6 co-fermentation
C5/C6 co-fermentation - No cell recycling due to solids presence.
Biomass
¹ Does not consider mineral impurities;² Composition for baled straw. Integral harvested straw moisture varies according to recovery fraction: 33.6 % for 50% recovery; 31.3% for 60% recovery; and 29.6% for 70% recovery;3 Energy cane composition includes its straw (100%). Source: Junqueira, 2015 CTBE/CNPEM
Biomass composition (wt %) ¹
Sugarcane stalks Sugarcane straw ² Energy cane3
Water 70.3% 15.0% 66.8%
Sucrose 14.0% 4.3% 8.1%
Reducing sugars 0.6% 0.2% 2.5%
Fibers 12.7% 77.9% 21.3%
Cellulose 6.0% 32.4% 10.0%
Hemicellulose 3.5% 24.8% 5.9%
Lignin 3.2% 20.6% 5.4%
Others 2.4% 2.6% 1.3%
Steam explosion pretreatmentShort term Medium term Long term
Temperature (°C) 190 200 210
Residence time (min) 15 10 5
Solids content (%) defined by the steam required totemperature
achieve reactor
Cellulose conversion to glucose (%) 0.5% 1.0% 1.0%
Cellulose conversion to glucose oligomers (%) 3.0% 3.0% 3.0%
Cellulose degradation to HMF (%) 1.5% 1.5% 1.5%
Xylan conversion to xylose (%) 30% 45% 60%
Xylan conversion to xylose oligomers (%) 30% 25% 20%
Xylan degradation to furfural (%) 10% 10% 10%
Lignin solubilization (%) 10% 10% 10%
Acetyl group conversion to acetic acid (%) 70% 80% 90%
Source: Junqueira, 2015 CTBE/CNPEM
9
Enzymatic HydrolysisShort term Medium term Long term
Temperature (°C) 50 50 65
Pressure (bar) 1.0 1.0 1.0
Residence time (h) 48 36 36
Solids content (%) 15 20 25
Cellulose conversion to glucose (%) 60% 70% 80%
Xylan conversion to xylose (%) 60% 70% 80%
Acetyl group conversion to acetic acid (%) 60% 70% 80%
Xylose oligomers to xylose (%) 60% 70% 80%
Source: Junqueira, 2015 CTBE/CNPEM
Fermentation
C6/C12 fermentation Short term Medium term Long term
Operational conditions same as 1G same as 1G same as 1G
C6/C12 conversion to ethanol (%)
88 90 90
Maximum alcoholic content (g/L)
70 85 85
Short term and “A” scenarios – Separated C5 fermentationDeoligomerization and C5 fermentation Short term Medium term Long term
Temperature (°C) 33 33 33
Residence time (h) 48 36 24
Xylose oligomers conversion to xylose (%) 80% 90% 90%
Glucose oligomers conversion to glucose (%) 80% 90% 90%
C6 conversion to ethanol (%) 90% 90% 90%
C5 conversion to ethanol (%) 80% 80% 85%
Maximum alcoholic content (g/L) 70 70 70
Cell Recycling (%) * 80% 90% 95%
Source: Junqueira, 2015 CTBE/CNPEM
Methodology of Identification of PatentsRelated only to EH
Methodology
Searching for a clear delimitation of the subject Using IPC Search: EH
Step 1
Key-words combination (Ten Groups- take Group 1 for instance
Group 1
biochemical + conversion + biomassbiochemical + conversion + biomass + lignocellulosicbiochemical + conversion + biomass + cellulosisbiochemical + conversion + biomass + hemicellulosisbiochemical + conversion + biomass + lignocellulosic + ethanolbiochemical + conversion + biomass + lignocellulosic + ethanol + fuel
biochemical + conversion + biomass
SubclassImportance(
%)Subgroups
C12P 25 C12P7/10 C12P7/06 C12P19/14 C12P19/00
C12N 18 C12N1/20 C12N15/29 C12N1/12 C12N1/14 C12N1/22
C10G 13 C10G3/00 C10G1/00 C10G1/08 C10G11/18
C10L 10 C10L5/44 C10L3/08
B01D 8 B01D1/14 B01D21/01 B01D25/00 B01D53/62 B01D53/78
C08H 7 C08H8/00
C10B 7 C10B49/22 C10B53/02
C13K 7 C13K1/02 C13K13/00
D21C 7 D21C1/02 D21C3/04
B01J 6 B01J8/18 B01J8/24 B01J29/40 B01J8/00 B01J8/32
Strongly concentrated on
those sub-classes
Selection of 44 IPC codes
MethodologySearching for EH (rigorous processes of convergence to the goal)
STEP 2 Selecting Patents in the Data Base Derwent Innovations Index (1970-2014)
“IP=(C12P-007/06 OR C12P-007/08 OR C12P-007/10 OR C12P-007/14 OR C12P-019/00 OR C12P-019/02 OR C12P-019/04 OR C12P-019/12 OR C12P-019/14 OR C12P-039/00 OR C12N-001/00 OR C12N-001/12 OR C12N-001/13 OR C12N-001/14 OR C12N-001/15 OR C12N-001/16 OR C12N-001/18 OR C12N-001/19 OR C12N-001/20 OR C12N-001/21 OR C12N-001/22 OR C12N-009/00 OR C12N-009/02 OR C12N-009/04 OR C12N-009/14 OR C12N-009/24 OR C12N-009/42 OR C12N-015/01 OR C12N-015/02 OR C12N-015/03 OR C12N-015/04 OR C12N-015/05 OR C12N-015/09 OR C12N-015/10 OR C12N-015/11 OR C12N-015/52 OR C12N-015/53 OR C12N-015/54 OR C12N-015/55 OR C12N-015/56 OR C12N-015/63 OR C12N-015/80 OR C12N-015/81 OR C12N-015/82)”
“biochemical conversion OR bioconversion OR hydrolysis OR saccharification OR biomass OR lignocellulose OR lignocellulosic OR cellulose OR cellulosic OR hemicellulose OR enzyme OR enzymes OR enzymatic OR ethanol OR bioethanol OR fuel OR biofuel”
“Acetivibrio OR Bacillus OR Bacteroides OR Cellulomonas OR Clostridium OR Erwinia OR Ruminococcus OR Streptomyces OR Microbispora OR Thermomonospora OR (genus AND bacterium) OR (genus AND bacteria) OR (genus AND bacteria*) OR (genera AND bacterium) OR (genera AND bacteria) OR (genera AND bacteria*) OR (specie AND bacterium) OR (specie AND bacteria) OR (specie AND bacteria*) OR (species AND bacterium) OR (species AND bacteria) OR (species AND bacteria*) OR (bacterium OR bacteria) OR Aspergillus OR Penicillium OR Phanerochaete OR Schizophyllum OR Sclerotinia OR Trichoderma OR (genus AND fungus) OR (genus AND fungi) OR (genus AND fung*) OR (genera AND fungus) OR (genera AND fungi) OR (genera AND fung*) OR (specie AND fungus) OR (specie AND fungi) OR (specie AND fung*) OR (species AND fungus) OR (species AND fungi) OR (species AND fung*) OR (fungus OR fungi)”
“swichgrass OR miscanthus OR prairie grass OR eucalyptus OR softwood OR hardwood OR willow OR poplar OR spruce OR loblolly pine OR pinus taeda OR cane bagasse OR bagasse OR corn stover OR stover OR husk OR stalk OR treetop OR treetops OR branch OR branches OR perennial grass OR perennial grasses OR residue OR residues OR waste OR forest”
IPC
Adventious terms
Microorg.
Raw materials
Methodology: EH
Step 2: Patents from Derwent Innovations Index (1970-2014)
Period Filterby
IPCs
Filter byAdventious
terms
By micro-organisms
By Raw materials
Crossing: micro versus raw material
1970-1979 673 268 89 44 9
1980-1989 10.566 3.852 1.368 499 168
1990-1999 35.553 8.831 2.634 1.360 454
2000-2009 98.852 29.832 9.364 6.758 2.311
2010-2014 48.012 15.827 5.631 3.266 1.176
Total 193.656 58.610 19.086 11.927 4.118
CHART1 – Number of Patetns by filter
Source: base Derwent Innovations Index, in <http://www.periodicos.capes.gov.br/>. Downloads in: 14, 15, 16, 19 e 20 May 2014.
Methodology: Identifying EH
STEP 3: Consulting a expert group: 304 patents were selected
1975-79 1980-84 1985-89 1990-94 1995-99 2000-04 2005-09 2010-140
20
40
60
80
100
120
140
2
13
512
24
45
79
124
By year of filling
Nu
mb
er o
f p
aten
ts
Preliminary Results
Coutry Filling Agent N. Pat.
1 USA
Genencor International Inc. 21
Danisco A/S 18
DuPont Nutrition Biosciences* 2
2 Denmark
Novozymes 19Novo Nordisk 11
Allopartis Biotechnologies Inc. 1
3 USA Codexis Inc. 11
4 GermanyVerenium Corp. 5Diversa Corp. 4Basf AG* 2
5 France IFP Energies Nouvelles 106 Canada Iogen Energy Corp. 97 USA Mascoma Corp. 88 GB BP Corp North America Inc. 8
9Netherlands DSM 6Netherlands Gist-brocades 2
10 USA Qteros Inc. 711 USA Dyadic International Inc. 7
12
USA MRIGlobal - Midwest Research Institute 5
USA Alliance for Sustainable Energy, LLC 1
USA Battelle Memorial Institute 1
CHART 2– Main Companies Filling Patents in EH field (1970-2010)
Applying the NK modelWhy to choose NK modelling ?
FRENKEN, K. A complexity approach to innovation networks. The case of the aircraft industry (1909 - 1997). Research Policy, v. 29, n. 2, p.257-272, Feb. 2000.
FRENKEN, K. Technological Innovation and Complexity Theory. Economics of Innovation and New Technology, v. 15, n. 2, p.137-155, 2006Koen Frenken (2006), Murakamy (2015)
Description of the Model
Methodology to apply NK model: mutual information indicators
First Category of information: Research Fields
Contents
1 Culture of Microorganisms – definition of parameters
2 OGM
3 Gene Expression in yeast/plants for expression of lignocellolitic Enzymes
4 Discovery of new microorganisms
5 Isolation, cloning and modification of genes that codifies enzymes
6 Changing aminoacids sequences
7 Preparing mix of Enzymes
Mutual Information Indicators(2) Microorganisms
(90 different genus of microorg).
Genus N. de pat Classification Species
1 Trichoderma 97 FungiT. reesei; T. viride; T. harzianum; T. longibrachiatum; T. asperellum; T. atroviride; T. cremeum; T. inhamatum; T. konilangbra; T. koningii; T. pseudokoningii; T. virens
2 Aspergillus 52 FungiA. niger; A. aculeatus; A. oryzae; A. fumigatus; A. tubingensis; A. terreus; A. usamii; A. awamori; A. japonicus; A. cellulolyticus; A. foetidus; A. saccharolyticus
3 Bacillus 37 BacteriaB. subtilis; B. licheniformis; B. halodurans; B. agaradhaerens; B. amyloliquefaciens; B. circulans; B. acidocaldarius; B. cereus; B. pumilus; B. stearothermophillus; B. velezensis
4 Penicillium 24 FungiP. funiculosum; P. brasilianum; P. verruculosum; P. janthinellum; P. decumbens; P. oxalicum; P. pinophilum; P. canescens; P. chrysogenum; P. citrinum; P. emersonii; P. funiculosum; P. griseofulvum; P. swollenin
5 Clostridium 20 Bacteria
C. phytofermentans; C. thermocellum; C. cellulolyticum; C. stercorarium; C. thermosulfurogenes; C. aldrichii; C. celerecrescens; C. cellobioparum; C. cellulosi; C. cellulovorans; C. chartatabidum; C. herbivorans; C. hungatei; C. josui;C. lentocellum; C. papyrosolvens; C. polysaccharolyticum; C. populeti; C. termitidis; C. thermocopriae
Mutual information Indicators
33 kinds of Lignocellulosic Enzymes
Enzymes N. de pat Group
1 endo-beta-1,4-glucanase 127 Celulase
2 endo-beta-1,4-xilanase 96 Hemicelulase
3 exo-beta-1,4-glucanase 92 Celulase
4 beta-glucosidase 79 Celulase
5 celulase 60 Celulase
Mutual information indicators
Formulas
Tri-dimensional
Bi-dimensionals
𝑇 ( 𝑋 ,𝑌 ,𝑍 )=∑𝑥=1
7
∑𝑦=1
83
∑𝑧=1
34
𝑝𝑥𝑦𝑧 log2(𝑝 𝑥𝑦𝑧
𝑝𝑥 .𝑝 𝑦 .𝑝𝑧
)
Results: values of T(X,Y,Z) of patentes, 1975-2013
Modelo NK Two dimensions T(X,Y) co-occurence
Modelo NK T(Y,Z) co-ocurrence
Dimension:content
definition of the parameters of the culture mean
application of the biotechnological techniques
Dimensionenzyme
cellulases (endo-beta-1,4-glucanase, exo-beta-1,4-glucanase and beta-glucosidase) and endo-beta-1,4-xylanase
TrichodermaAspergillus
Others
Bacillus
Penicillium
others
others
Chrysosporium
Humicola
Dimension:
micro-organism
Clostridium
Myceliophthora
StreptomycesTalaromyces
Technology trajectories in 2G
Conclusions
A substantial growth in the number of patents, even before having a convergence of Technologies,
Few corporations fiiled a considerable part of patentes, showing the importance of cumulativeness and technological tradition (DUPONT e NOVOZYMES)
Merger and acquisitions
Barriers to new entrants
Brazil: only one patent: Petrobrás (2009)????????
Questions
NK model allow us to detect the main combinations of the three: reinforced trajectories
Pre-paradigmatic stage: Windows of opportunities?
There is some convinience to guide research and patenting to a certain microorganisms, Enzymes and even methods?
Direction of TT: accelerating convergence of Technologies to a standart packges?
Supply from Global Corporations
If it is so, Why researcher in Brazil is focused on EH?