Abstract:
BACKGROUND:Bioethanol produced by the yeast Saccharomyces cerevisiae is currently one of the most promising alternatives to conventional transport fuels. Lignocellulosic hemicelluloses obtained after hydrothermal pretreatment are important feedstock for bioethanol production. However, hemicellulosic materials cannot be directly fermented by yeast: xylan backbone of hemicelluloses must first be hydrolyzed by heterologous hemicellulases to release xylose, and the yeast must then ferment xylose in the presence of fermentation inhibitors generated during the pretreatment. RESULTS:A GIN11/FRT-based multiple-gene integration system was developed for introducing multiple functions into the recombinant S. cerevisiae strains engineered with the xylose metabolic pathway. Antibiotic markers were efficiently recycled by a novel counter selection strategy using galactose-induced expression of both FLP recombinase gene and GIN11 flanked by FLP recombinase recognition target (FRT) sequences. Nine genes were functionally expressed in an industrial diploid strain of S. cerevisiae: endoxylanase gene from Trichoderma reesei, xylosidase gene from Aspergillus oryzae, β-glucosidase gene from Aspergillus aculeatus, xylose reductase and xylitol dehydrogenase genes from Scheffersomyces stipitis, and XKS1, TAL1, FDH1 and ADH1 variant from S. cerevisiae. The genes were introduced using the homozygous integration system and afforded hemicellulolytic, xylose-assimilating and inhibitor-tolerant abilities to the strain. The engineered yeast strain demonstrated 2.7-fold higher ethanol titer from hemicellulosic material than a xylose-assimilating yeast strain. Furthermore, hemicellulolytic enzymes displayed on the yeast cell surface hydrolyzed hemicelluloses that were not hydrolyzed by a commercial enzyme, leading to increased sugar utilization for improved ethanol production. CONCLUSIONS:The multifunctional yeast strain, developed using a GIN11/FRT-based marker recycling system, achieved direct conversion of hemicellulosic biomass to ethanol without the addition of exogenous hemicellulolytic enzymes. No detoxification processes were required. The multiple-gene integration technique is a powerful approach for introducing and improving the biomass fermentation ability of industrial diploid S. cerevisiae strains.
journal_name
Microb Cell Factjournal_title
Microbial cell factoriesauthors
Hasunuma T,Hori Y,Sakamoto T,Ochiai M,Hatanaka H,Kondo Adoi
10.1186/s12934-014-0145-9subject
Has Abstractpub_date
2014-10-12 00:00:00pages
145issn
1475-2859pii
s12934-014-0145-9journal_volume
13pub_type
杂志文章abstract::Methylotrophic yeasts such as Candida boidinii, Hansenula polymorpha, Pichia methanolica and Pichia pastoris are an emerging group of eukaryotic hosts for recombinant protein production with an ever increasing number of applications during the last 30 years. Their applications are linked to the use of strong methanol-...
journal_title:Microbial cell factories
pub_type: 杂志文章
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更新日期:2006-12-14 00:00:00
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journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-015-0378-2
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journal_title:Microbial cell factories
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journal_title:Microbial cell factories
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abstract::Lactococcus lactis has progressed a long way since its discovery and initial use in dairy product fermentation, to its present biotechnological applications in genetic engineering for the production of various recombinant proteins and metabolites that transcends the heterologous species barrier. Key desirable features...
journal_title:Microbial cell factories
pub_type: 杂志文章,评审
doi:10.1186/s12934-017-0669-x
更新日期:2017-04-04 00:00:00
abstract:BACKGROUND:Recombinant protein expression in bacteria often leads to the formation of intracellular insoluble protein deposits, a major bottleneck for the production of soluble and active products. However, in recent years, these bacterial protein aggregates, commonly known as inclusion bodies (IBs), have been shown to...
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pub_type: 杂志文章
doi:10.1186/s12934-020-01375-4
更新日期:2020-06-01 00:00:00
abstract:BACKGROUND:A reliable quenching and metabolite extraction method has been developed for Lactobacillus plantarum. The energy charge value was used as a critical indicator for fixation of metabolism. RESULTS:Four different aqueous quenching solutions, all containing 60% of methanol, were compared for their efficiency. O...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-6-27
更新日期:2007-08-20 00:00:00
abstract:BACKGROUND:The production of high yields of recombinant proteins is an enduring bottleneck in the post-genomic sciences that has yet to be addressed in a truly rational manner. Typically eukaryotic protein production experiments have relied on varying expression construct cassettes such as promoters and tags, or cultur...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-8-10
更新日期:2009-01-29 00:00:00
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journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-016-0450-6
更新日期:2016-03-15 00:00:00
abstract:BACKGROUND:The filamentous fungus T. reesei effectively degrades cellulose and is known to produce various cellulolytic enzymes such as beta-glucosidase, endoglucanase, and cellobiohydrolase. The expression levels of each cellulase are controlled simultaneously, and their ratios and synergetic effects are important for...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-9-32
更新日期:2010-05-14 00:00:00
abstract:BACKGROUND:The CRISPR/Cas9 system is a powerful tool for genome editing, in which the sgRNA binds and guides the Cas9 protein for the sequence-specific cleavage. The protocol is employable in different organisms, but is often limited by cell damage due to the endonuclease activity of the introduced Cas9 and the potenti...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-017-0758-x
更新日期:2017-08-16 00:00:00
abstract:BACKGROUND:High-throughput methods are widely-used for strain screening effectively resulting in binary information regarding high or low productivity. Nevertheless achieving quantitative and scalable parameters for fast bioprocess development is much more challenging, especially for heterologous protein production. He...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-11-144
更新日期:2012-10-31 00:00:00
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pub_type: 杂志文章
doi:10.1186/1475-2859-10-4
更新日期:2011-01-28 00:00:00
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journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-6-30
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更新日期:2017-11-02 00:00:00
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journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-7-26
更新日期:2008-08-07 00:00:00
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journal_title:Microbial cell factories
pub_type: 杂志文章
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更新日期:2015-09-15 00:00:00
abstract:BACKGROUND:In Saccharomyces cerevisiae, alpha-glucosidase (maltase) is a key enzyme in maltose metabolism. In addition, the overexpression of the alpha-glucosidase-encoding gene MAL62 has been shown to increase the freezing tolerance of yeast in lean dough. However, its cryoprotection mechanism is still not clear. RES...
journal_title:Microbial cell factories
pub_type: 杂志文章
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更新日期:2020-10-19 00:00:00
abstract:BACKGROUND:Oil palm empty fruit bunch (EFB) is a lignocellulosic waste produced in palm oil industry. EFB mainly consists of cellulose, hemicellulose (mainly xylan) and lignin and has a great potential to be reused. Converting EFB to fermentable sugars and value-added chemicals is a much better choice than treating EFB...
journal_title:Microbial cell factories
pub_type: 杂志文章
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更新日期:2014-11-11 00:00:00
abstract:BACKGROUND:Catalase (EC 1.11.1.6) is one of the important industrial enzyme employed in diagnostic and analytical methods in the form of biomarkers and biosensors in addition to their enormous applications in textile, paper, food and pharmaceutical sectors. The present study demonstrates the utility of a newly isolated...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-016-0410-1
更新日期:2016-01-14 00:00:00
abstract:BACKGROUND:(2S)-Pinocembrin is a chiral flavanone with versatile pharmacological and biological activities. Its health-promoting effects have spurred on research effects on the microbial production of (2S)-pinocembrin. However, an often-overlooked salient feature in the analysis of microbial (2S)-pinocembrin is its chi...
journal_title:Microbial cell factories
pub_type: 杂志文章
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更新日期:2016-02-04 00:00:00
abstract:BACKGROUND:Bioethanol is one of the most representative eco-friendly fuels developed to replace the non-renewable fossil fuels and is the most successful commercially available bio-conversion technology till date. With the availability of inexpensive carbon sources, such as cellulosic biomass, bioethanol production has...
journal_title:Microbial cell factories
pub_type: 杂志文章
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更新日期:2019-10-10 00:00:00
abstract:BACKGROUND:Trypsin has many applications in food and pharmaceutical manufacturing. Although commercial trypsin is usually extracted from porcine pancreas, this source carries the risks of infectivity and immunogenicity. Microbial Streptomyces griseus trypsin (SGT) is a prime alternative because it possesses efficient h...
journal_title:Microbial cell factories
pub_type: 杂志文章
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更新日期:2020-04-13 00:00:00