Cellulosic ethanol

From revolutionary consolidated bioprocessing idea to proof of concept

W. H. Van Zyl, Riaan Den Haan, Shaunita H. Rose, Daniel C. La Grange, Ronel van Rooyen, John E. Mcbride, Lee R. Lynd, Marja Ilmen, Merja Penttilä

Research output: Contribution to conferenceOther conference contributionScientific

Abstract

Four biological events occur during conversion of lignocellulose to ethanol via processes featuring enzymatic hydrolysis: production of glycosyl hydrolase enzymes (cellulases and hemicellulases), hydrolysis of the polysaccharides present in pretreated biomass, fermentation of hexose sugars, and fermentation of pentose sugars. Consolidation of these events in a single process step via a cellulolytic microorganism(s) is a promising approach to low-cost conversion of lignocellulose to fuels and chemicals. One strategy for developing a microorganism capable of such consolidated bioprocessing involves engineering the non-cellulolytic yeast Saccharomyces cerevisiae so that it expresses a heterologous cellulase system enabling crystalline cellulose utilization.

We developed the fundamental principles behind consolidated bioprocessing as a microbial phenomenon and illustrated proof of concept in the laboratory through the successful expression of the three major cellulase activities (β‑glucosidase, β‑endo-glucanase and cellobiohydrolase) in S. cerevisiae. We have enabled this yeast to grow on cellobiose, amorphous cellulose (PASC) and crystalline cellulose (BMCC) through the combinational expression of a β‑glucosidase, endo-1,4-β-glucanase and cellobiohydrolase genes.

Recently, as part of an international team funded by Mascoma Corporation in the USA, we accelerated the research effort and transferred appropriate genes to commercial yeast strains. Expression levels of CBH genes have been improved up to 2500-fold and collectively we could demonstrate with proprietary CBP-yeast strains efficient conversion of unwashed pretreated hardwoods to ethanol with a 2.5-fold reduction in cellulase loading at pilot scale, which represents a major reduction in operating costs for the commercial production of cellulosic ethanol. Conversion of 18% w/w waste paper sludge to ethanol was demonstrated with minimal external enzyme addition. These breakthroughs bring the application of CBP at commercial scale closer than anticipated as recent as a year ago.
Original languageEnglish
Publication statusPublished - 2010
EventInternational Workshop on Wood Biorefinery and Tree Biotechnology - Örnsköldsvik, Sweden
Duration: 21 Jun 201023 Jun 2010

Workshop

WorkshopInternational Workshop on Wood Biorefinery and Tree Biotechnology
CountrySweden
CityÖrnsköldsvik
Period21/06/1023/06/10

Fingerprint

bioprocessing
bioethanol
endo-1,4-beta-glucanase
cellulose 1,4-beta-cellobiosidase
yeasts
lignocellulose
glucosidases
cellulose
cellulases
ethanol
Saccharomyces cerevisiae
cellulolytic microorganisms
fermentation
waste paper
pulp and paper sludge
sugars
cellobiose
operating costs
genes
pentoses

Cite this

Van Zyl, W. H., Den Haan, R., Rose, S. H., La Grange, D. C., van Rooyen, R., Mcbride, J. E., ... Penttilä, M. (2010). Cellulosic ethanol: From revolutionary consolidated bioprocessing idea to proof of concept. International Workshop on Wood Biorefinery and Tree Biotechnology, Örnsköldsvik, Sweden.
Van Zyl, W. H. ; Den Haan, Riaan ; Rose, Shaunita H. ; La Grange, Daniel C. ; van Rooyen, Ronel ; Mcbride, John E. ; Lynd, Lee R. ; Ilmen, Marja ; Penttilä, Merja. / Cellulosic ethanol : From revolutionary consolidated bioprocessing idea to proof of concept. International Workshop on Wood Biorefinery and Tree Biotechnology, Örnsköldsvik, Sweden.
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title = "Cellulosic ethanol: From revolutionary consolidated bioprocessing idea to proof of concept",
abstract = "Four biological events occur during conversion of lignocellulose to ethanol via processes featuring enzymatic hydrolysis: production of glycosyl hydrolase enzymes (cellulases and hemicellulases), hydrolysis of the polysaccharides present in pretreated biomass, fermentation of hexose sugars, and fermentation of pentose sugars. Consolidation of these events in a single process step via a cellulolytic microorganism(s) is a promising approach to low-cost conversion of lignocellulose to fuels and chemicals. One strategy for developing a microorganism capable of such consolidated bioprocessing involves engineering the non-cellulolytic yeast Saccharomyces cerevisiae so that it expresses a heterologous cellulase system enabling crystalline cellulose utilization.We developed the fundamental principles behind consolidated bioprocessing as a microbial phenomenon and illustrated proof of concept in the laboratory through the successful expression of the three major cellulase activities (β‑glucosidase, β‑endo-glucanase and cellobiohydrolase) in S. cerevisiae. We have enabled this yeast to grow on cellobiose, amorphous cellulose (PASC) and crystalline cellulose (BMCC) through the combinational expression of a β‑glucosidase, endo-1,4-β-glucanase and cellobiohydrolase genes.Recently, as part of an international team funded by Mascoma Corporation in the USA, we accelerated the research effort and transferred appropriate genes to commercial yeast strains. Expression levels of CBH genes have been improved up to 2500-fold and collectively we could demonstrate with proprietary CBP-yeast strains efficient conversion of unwashed pretreated hardwoods to ethanol with a 2.5-fold reduction in cellulase loading at pilot scale, which represents a major reduction in operating costs for the commercial production of cellulosic ethanol. Conversion of 18{\%} w/w waste paper sludge to ethanol was demonstrated with minimal external enzyme addition. These breakthroughs bring the application of CBP at commercial scale closer than anticipated as recent as a year ago.",
author = "{Van Zyl}, {W. H.} and {Den Haan}, Riaan and Rose, {Shaunita H.} and {La Grange}, {Daniel C.} and {van Rooyen}, Ronel and Mcbride, {John E.} and Lynd, {Lee R.} and Marja Ilmen and Merja Penttil{\"a}",
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Van Zyl, WH, Den Haan, R, Rose, SH, La Grange, DC, van Rooyen, R, Mcbride, JE, Lynd, LR, Ilmen, M & Penttilä, M 2010, 'Cellulosic ethanol: From revolutionary consolidated bioprocessing idea to proof of concept' International Workshop on Wood Biorefinery and Tree Biotechnology, Örnsköldsvik, Sweden, 21/06/10 - 23/06/10, .

Cellulosic ethanol : From revolutionary consolidated bioprocessing idea to proof of concept. / Van Zyl, W. H.; Den Haan, Riaan; Rose, Shaunita H.; La Grange, Daniel C.; van Rooyen, Ronel; Mcbride, John E.; Lynd, Lee R.; Ilmen, Marja; Penttilä, Merja.

2010. International Workshop on Wood Biorefinery and Tree Biotechnology, Örnsköldsvik, Sweden.

Research output: Contribution to conferenceOther conference contributionScientific

TY - CONF

T1 - Cellulosic ethanol

T2 - From revolutionary consolidated bioprocessing idea to proof of concept

AU - Van Zyl, W. H.

AU - Den Haan, Riaan

AU - Rose, Shaunita H.

AU - La Grange, Daniel C.

AU - van Rooyen, Ronel

AU - Mcbride, John E.

AU - Lynd, Lee R.

AU - Ilmen, Marja

AU - Penttilä, Merja

N1 - CO:U University of Stellenbosch, Stellenbosch, South Africa CO:U Thayer School of Engineering, Hanover, NH, USA CA2: TK402 CA2: TK400

PY - 2010

Y1 - 2010

N2 - Four biological events occur during conversion of lignocellulose to ethanol via processes featuring enzymatic hydrolysis: production of glycosyl hydrolase enzymes (cellulases and hemicellulases), hydrolysis of the polysaccharides present in pretreated biomass, fermentation of hexose sugars, and fermentation of pentose sugars. Consolidation of these events in a single process step via a cellulolytic microorganism(s) is a promising approach to low-cost conversion of lignocellulose to fuels and chemicals. One strategy for developing a microorganism capable of such consolidated bioprocessing involves engineering the non-cellulolytic yeast Saccharomyces cerevisiae so that it expresses a heterologous cellulase system enabling crystalline cellulose utilization.We developed the fundamental principles behind consolidated bioprocessing as a microbial phenomenon and illustrated proof of concept in the laboratory through the successful expression of the three major cellulase activities (β‑glucosidase, β‑endo-glucanase and cellobiohydrolase) in S. cerevisiae. We have enabled this yeast to grow on cellobiose, amorphous cellulose (PASC) and crystalline cellulose (BMCC) through the combinational expression of a β‑glucosidase, endo-1,4-β-glucanase and cellobiohydrolase genes.Recently, as part of an international team funded by Mascoma Corporation in the USA, we accelerated the research effort and transferred appropriate genes to commercial yeast strains. Expression levels of CBH genes have been improved up to 2500-fold and collectively we could demonstrate with proprietary CBP-yeast strains efficient conversion of unwashed pretreated hardwoods to ethanol with a 2.5-fold reduction in cellulase loading at pilot scale, which represents a major reduction in operating costs for the commercial production of cellulosic ethanol. Conversion of 18% w/w waste paper sludge to ethanol was demonstrated with minimal external enzyme addition. These breakthroughs bring the application of CBP at commercial scale closer than anticipated as recent as a year ago.

AB - Four biological events occur during conversion of lignocellulose to ethanol via processes featuring enzymatic hydrolysis: production of glycosyl hydrolase enzymes (cellulases and hemicellulases), hydrolysis of the polysaccharides present in pretreated biomass, fermentation of hexose sugars, and fermentation of pentose sugars. Consolidation of these events in a single process step via a cellulolytic microorganism(s) is a promising approach to low-cost conversion of lignocellulose to fuels and chemicals. One strategy for developing a microorganism capable of such consolidated bioprocessing involves engineering the non-cellulolytic yeast Saccharomyces cerevisiae so that it expresses a heterologous cellulase system enabling crystalline cellulose utilization.We developed the fundamental principles behind consolidated bioprocessing as a microbial phenomenon and illustrated proof of concept in the laboratory through the successful expression of the three major cellulase activities (β‑glucosidase, β‑endo-glucanase and cellobiohydrolase) in S. cerevisiae. We have enabled this yeast to grow on cellobiose, amorphous cellulose (PASC) and crystalline cellulose (BMCC) through the combinational expression of a β‑glucosidase, endo-1,4-β-glucanase and cellobiohydrolase genes.Recently, as part of an international team funded by Mascoma Corporation in the USA, we accelerated the research effort and transferred appropriate genes to commercial yeast strains. Expression levels of CBH genes have been improved up to 2500-fold and collectively we could demonstrate with proprietary CBP-yeast strains efficient conversion of unwashed pretreated hardwoods to ethanol with a 2.5-fold reduction in cellulase loading at pilot scale, which represents a major reduction in operating costs for the commercial production of cellulosic ethanol. Conversion of 18% w/w waste paper sludge to ethanol was demonstrated with minimal external enzyme addition. These breakthroughs bring the application of CBP at commercial scale closer than anticipated as recent as a year ago.

UR - http://bioenfapesp.org/images/stories/BIOEN_Workshop_October2010/Emile-Van-Zyl.pdf

M3 - Other conference contribution

ER -

Van Zyl WH, Den Haan R, Rose SH, La Grange DC, van Rooyen R, Mcbride JE et al. Cellulosic ethanol: From revolutionary consolidated bioprocessing idea to proof of concept. 2010. International Workshop on Wood Biorefinery and Tree Biotechnology, Örnsköldsvik, Sweden.