Direct and Efficient Production of Ethanol from Cellulosic Material with a Yeast Strain Displaying Cellulolytic Enzymes

Fujita, Yasuya; Takahashi, Shouji; Ueda, Mitsuyoshi; Tanaka, Atsuo; Okada, Hirofumi; Morikawa, Yasushi; Kawaguchi, Takashi; Arai, Motoo; Fukuda, Hideki; Kondo, Akihiko

doi:10.1128/aem.68.10.5136-5141.2002

Cited by 208 publications

(96 citation statements)

References 20 publications

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“…To facilitate direct ethanol fermentation from cellulose, we previously developed yeast strains displaying cellulolytic enzymes on the cell surface [5,10]. In this study, to increase ethanol productivity, we optimized the cellulase expression system.…”

Section: Research Articlementioning

confidence: 99%

“…The plasmid for cell surface display of the T. reesei EGII gene was constructed as follows. The DNA fragment encoding the secretion signal sequence from R. oryzae, the EGII gene, and the 3′ half of α-agglutinin was prepared by PCR using pEG23u31H6 [10] with the following primers: 5′-CATGCTAGCATGCAACTGTTCAATTTGCC ATTGAAAG-3′ and 5′-TCCCCCCGGGTTTGATT ATGTTCTTTCTATTTGAATGAGATATG-3′. The amplified fragment was digested with NheI and XmaI and ligated into pGK404 [15].…”

Section: Plasmid Constructionmentioning

confidence: 99%

“…BGL activity was determined using p-nitrophenyl-β-D-glucopyranoside as the substrate according to a previously described method [10].…”

Section: Enzymatic Assaysmentioning

confidence: 99%

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Ethanol production from cellulosic materials using cellulase‐expressing yeast

Yanase

Yamada

Kaneko³

et al. 2010

Biotechnology Journal

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We demonstrate direct ethanol fermentation from amorphous cellulose using cellulase-coexpressing yeast. Endoglucanases (EG) and cellobiohydrolases (CBH) from Trichoderma reesei, and β-glucosidases (BGL) from Aspergillus aculeatus were integrated into genomes of the yeast strain Saccharomyces cerevisiae MT8-1. BGL was displayed on the yeast cell surface and both EG and CBH were secreted or displayed on the cell surface. All enzymes were successfully expressed on the cell surface or in culture supernatants in their active forms, and cellulose degradation was increased 3-to 5-fold by co-expressing EG and CBH. Direct ethanol fermentation from 10 g/L phosphoric acid swollen cellulose (PASC) was also carried out using EG-, CBH-, and BGL-co-expressing yeast. The ethanol yield was 2.1 g/L for EG-, CBH-, and BGL-displaying yeast, which was higher than that of EG-and CBH-secreting yeast (1.6 g/L ethanol). Our results show that cell surface display is more suitable for direct ethanol fermentation from cellulose.

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Section: Research Articlementioning

confidence: 99%

Section: Plasmid Constructionmentioning

confidence: 99%

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Ethanol production from cellulosic materials using cellulase‐expressing yeast

Yanase

Yamada

Kaneko³

et al. 2010

Biotechnology Journal

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“…None of these recombinant strains could directly utilize cellulose, as complete cellulose hydrolysis required synergistic action of at least three main cellulases (Cho et al, 2004;Mingardon et al, 2005;Arai et al, 2007). Fujita et al (2002Fujita et al ( , 2004 reported co-expression and surface display of cellulases in S. cerevisiae. The recombinant strain displaying three EG2 and CBH2 (derived from T. reesei), and the β-glucosidase (derived from Aspergillus aculeatus) enzymes could directly convert 10 g/l phosphoric acid swollen cellulose to approximately 3g/l ethanol.…”

Section: -Yeast Surface Display (Non-complexed Cellulase Systems Celmentioning

confidence: 99%

Advances in consolidated bioprocessing systems for bioethanol and butanol production from biomass: a comprehensive review

2015

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HIGHLIGHTS Various CBP strategies have been discussed. Recently, lignocellulosic biomass as the most abundant renewable resource has been widely considered for bioalcohols production. However, the complex structure of lignocelluloses requires a multi-step process which is costly and time consuming. Although, several bioprocessing approaches have been developed for pretreatment, saccharification and fermentation, bioalcohols production from lignocelluloses is still limited because of the economic infeasibility of these technologies. This cost constraint could be overcome by designing and constructing robust cellulolytic and bioalcohols producing microbes and by using them in a consolidated bioprocessing (CBP) system. This paper comprehensively reviews potentials, recent advances and challenges faced in CBP systems for efficient bioalcohols (ethanol and butanol) production from lignocellulosic and starchy biomass. The CBP strategies include using native single strains with cellulytic and alcohol production activities, microbial co-cultures containing both cellulytic and ethanologenic microorganisms, and genetic engineering of cellulytic microorganisms to be alcohol-producing or alcohol producing microorganisms to be cellulytic. Moreover, high-throughput techniques, such as metagenomics, metatranscriptomics, next generation sequencing and synthetic biology developed to explore novel microorganisms and powerful enzymes with high activity, thermostability and pH stability are also discussed. Currently, the CBP technology is in its infant stage, and ideal microorganisms and/or conditions at industrial scale are yet to be introduced. So, it is essential to bring into attention all barriers faced and take advantage of all the experiences gained to achieve a high-yield and low-cost CBP process.

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“…A recent paper comparing cellobiose usage by surface expressed versus secreted β-glucosidase demonstrated that expression on the yeast cell surface stabilized and increased specific activity of the enzyme [36]. Fujita et al, [37] reported co-display of endoglucanase II and β-glucosidase on the surface of S. cerevisiae MT8-1. The strain was able to grow on β-glucan as the sole carbon source and produced 16.5 g/l of ethanol without pretreatment, whereas the strains without the co-displayed enzymes did not grow.…”

Section: Surface-display For Biofuels Productionmentioning

confidence: 99%

Versatile microbial surface-display for environmental remediation and biofuels production

Wu¹,

Mulchandani²,

Chen³

2008

Trends in Microbiology

101

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Surface display is a powerful technique that utilizes natural microbial functional components to express proteins or peptides on the cell exterior. Since the reporting of the first surface-display system in the mid-1980s, a variety of new systems have been reported for yeast, Gram-positive and Gram-negative bacteria. Non-conventional display methods are emerging, eliminating the generation of genetically modified microorganisms. Cells with surface display are used as biocatalysts, biosorbents and biostimulants. Microbial cell-surface display has proven to be extremely important for numerous applications ranging from combinatorial library screening and protein engineering to bioremediation and biofuels production.3

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Direct and Efficient Production of Ethanol from Cellulosic Material with a Yeast Strain Displaying Cellulolytic Enzymes

Cited by 208 publications

References 20 publications

Ethanol production from cellulosic materials using cellulase‐expressing yeast

Ethanol production from cellulosic materials using cellulase‐expressing yeast

Advances in consolidated bioprocessing systems for bioethanol and butanol production from biomass: a comprehensive review

Versatile microbial surface-display for environmental remediation and biofuels production

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