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Self-surface assembly of cellulosomes with two miniscaffoldins on Saccharomyces cerevisiae for cellulosic ethanol production

Proceedings of the National Academy of Sciences - PNAS, 2012-08, Vol.109 (33), p.13260-13265 [Peer Reviewed Journal]

copyright © 1993-2008 National Academy of Sciences of the United States of America ;Copyright National Academy of Sciences Aug 14, 2012 ;ISSN: 0027-8424 ;EISSN: 1091-6490 ;DOI: 10.1073/pnas.1209856109 ;PMID: 22853950

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Title:
Self-surface assembly of cellulosomes with two miniscaffoldins on Saccharomyces cerevisiae for cellulosic ethanol production
Author: Fan, Li-Hai ; Zhang, Zi-Jian ; Yu, Xiao-Yu ; Xue, Ya-Xu ; Tan, Tian-Wei
Subjects: beta-glucosidase ; Bioethanol ; Biological Sciences ; carboxymethylcellulose ; Cell Membrane - metabolism ; Cellulose ; Cellulose - metabolism ; cellulose 1,4-beta-cellobiosidase ; Cellulosic ethanol ; Cellulosomes ; Cellulosomes - metabolism ; Cloning ; Clostridium cellulolyticum ; endo-1,4-beta-glucanase ; Enzymes ; Ethanol ; Ethanol - metabolism ; ethanol fermentation ; ethanol production ; Fermentation ; Flow Cytometry ; fluorescence microscopy ; Fluorescent Antibody Technique ; Hydrolysis ; Models, Biological ; Product displays ; saccharification ; Saccharomyces cerevisiae ; Saccharomyces cerevisiae - cytology ; Saccharomyces cerevisiae - enzymology ; Saccharomyces cerevisiae - metabolism ; Time Factors ; Yeast ; Yeasts
Is Part Of: Proceedings of the National Academy of Sciences - PNAS, 2012-08, Vol.109 (33), p.13260-13265
Description: Yeast to directly convert cellulose and, especially, the microcrystalline cellulose into bioethanol, was engineered through display of minicellulosomes on the cell surface of Saccharomyces cerevisiae . The construction and cell surface attachment of cellulosomes were accomplished with two individual miniscaffoldins to increase the display level. All of the cellulases including a celCCA (endoglucanase), a celCCE (cellobiohydrolase), and a Ccel_2454 (β-glucosidase) were cloned from Clostridium cellulolyticum , ensuring the thermal compatibility between cellulose hydrolysis and yeast fermentation. Cellulases and one of miniscaffoldins were secreted by α-factor; thus, the assembly and attachment to anchoring miniscaffoldin were accomplished extracellularly. Immunofluorescence microscopy, flow cytometric analysis (FACS), and cellulosic ethanol fermentation confirmed the successful display of such complex on the yeast surface. Enzyme–enzyme synergy, enzyme-proximity synergy, and cellulose–enzyme–cell synergy were analyzed, and the length of anchoring miniscaffoldin was optimized. The engineered S. cerevisiae was applied in fermentation of carboxymethyl cellulose (CMC), phosphoric acid-swollen cellulose (PASC), or Avicel. It showed a significant hydrolytic activity toward microcrystalline cellulose, with an ethanol titer of 1,412 mg/L. This indicates that simultaneous saccharification and fermentation of crystalline cellulose to ethanol can be accomplished by the yeast, engineered with minicellulosome.
Publisher: United States: National Academy of Sciences
Language: English
Identifier: ISSN: 0027-8424
EISSN: 1091-6490
DOI: 10.1073/pnas.1209856109
PMID: 22853950
Source: PubMed Central (Open access)
Geneva Foundation Free Medical Journals at publisher websites
MEDLINE

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Self-surface assembly of cellulosomes with two miniscaffoldins on Saccharomyces cerevisiae for cellulosic ethanol production

copyright © 1993-2008 National Academy of Sciences of the United States of America ;Copyright National Academy of Sciences Aug 14, 2012 ;ISSN: 0027-8424 ;EISSN: 1091-6490 ;DOI: 10.1073/pnas.1209856109 ;PMID: 22853950

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