Construction of thermotolerant yeast expressing thermostable cellulase genes

Hong, Jiong; Wang, Yonghong; Kumagai, Hiroo; Tamaki, Hiroyuki

doi:10.1016/j.jbiotec.2007.03.008

Cited by 131 publications

(79 citation statements)

References 39 publications

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“…In this study, to increase ethanol productivity, we optimized the cellulase expression system. We utilized a genome integrated expression system for stable expression of cellulases [11] and the PGK promoter for overexpression [12,13]. Using EG-, CBH-, and BGL-displaying yeast, direct ethanol fermentation from amorphous cellulose was demonstrated.…”

Section: Research Articlementioning

confidence: 99%

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%

Ethanol production from cellulosic materials using cellulase‐expressing yeast

Yanase

Yamada

Kaneko³

et al. 2010

Biotechnology Journal

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“…Xylanase activity was increase up to 3.3 times more than free enzymes after 72 h hydrolysis Srikrishnan et al, 2013 Cellobiose or CMC (100 g/l) K. marxianus Thermoascus aurantiacus cellulase genes, including cellobiohydrolase 1 (cel7A), endoglucanase 1 (cel5A) and β-glucosidase (bgl3A) genes Secretion 43.4 g/l ethanol Hong et al 2007 .…”

Section: Xylanosomesmentioning

confidence: 99%

“…Previously, three Thermoascus aurantiacus cellulase genes, including cellobiohydrolase 1 (cel7A), endoglucanase 1 (cel5A) and β-glucosidase (bgl3A), were expressed in K. marxianus strains. The recombinant strain could efficiently grow in synthetic media containing cellobiose or CMC as sole carbon source, and produced 43.4 g/l ethanol from 100 g/l cellobiose in 24 h at 45°C (Hong et al 2007). Yanase et al (2010) used cell surface technology to develop a new recombinant strain of K. marxianus with cellulytic activities for CBP ethanol production.…”

Section: Kluyveromyces Marxianusmentioning

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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“…For example, the thermotolerant Kluyveromyces marxianus was used in a number of SSF studies in which cellulases were added exogenously [34][35][36][37]. The strain was subsequently engineered to express three thermostable cellulases, endowing it with the ability to grow at 45°C on both cellobiose and carboxymethyl cellulose and to ferment cellobiose to ethanol [38]. The multi-stress tolerant Issatchenka orientalis was also successfully engineered for recombinant cellulase expression.…”

Section: Recombinant Cellulase Expression In Other Yeast Speciesmentioning

confidence: 99%