Effective saccharification of kraft pulp by using a cellulase cocktail prepared from genetically engineered <i>Aspergillus oryzae</i>

Yamada, R.; Yoshie, Toshihide; Sakai, Shoji; Wakai, Satoshi; Asai-Nakashima, Nanami; Okazaki, Fumiyoshi; Ogino, Chiaki; Hisada, Hiromoto; Tsutsumi, Hiroko; Hata, Yutaka; Kondo, Akihiko

doi:10.1080/09168451.2015.1006568

Cited by 9 publications

(3 citation statements)

References 13 publications

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“…The higher lignin content of the mechanical pulps may explain why their mechanical strength was not affected by the treatment. Analysis of these paper properties corroborates previous studies of simple sugars release by hydrolysis of paper pulp and confirms that Kraft pulps are more susceptible to enzymatic treatments [ 47 , 50 , 86 , 87 ]. For applications where strength properties are very important, such combination pulp-enzymatic treatments (Kraft pulps treated with cellulase mixtures) would be deleterious.…”

Section: Resultssupporting

confidence: 86%

Predicting the most appropriate wood biomass for selected industrial applications: comparison of wood, pulping, and enzymatic treatments using fluorescent-tagged carbohydrate-binding modules

Bombeck

Khatri

Meddeb-Mouelhi

et al. 2017

Biotechnol Biofuels

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BackgroundLignocellulosic biomass will progressively become the main source of carbon for a number of products as the Earth’s oil reservoirs disappear. Technology for conversion of wood fiber into bioproducts (wood biorefining) continues to flourish, and access to reliable methods for monitoring modification of such fibers is becoming an important issue. Recently, we developed a simple, rapid approach for detecting four different types of polymer on the surface of wood fibers. Named fluorescent-tagged carbohydrate-binding module (FTCM), this method is based on the fluorescence signal from carbohydrate-binding modules-based probes designed to recognize specific polymers such as crystalline cellulose, amorphous cellulose, xylan, and mannan.ResultsHere we used FTCM to characterize pulps made from softwood and hardwood that were prepared using Kraft or chemical-thermo-mechanical pulping. Comparison of chemical analysis (NREL protocol) and FTCM revealed that FTCM results were consistent with chemical analysis of the hemicellulose composition of both hardwood and softwood samples. Kraft pulping increased the difference between softwood and hardwood surface mannans, and increased xylan exposure. This suggests that Kraft pulping leads to exposure of xylan after removal of both lignin and mannan. Impact of enzyme cocktails from Trichoderma reesei (Celluclast 1.5L) and from Aspergillus sp. (Carezyme 1000L) was investigated by analysis of hydrolyzed sugars and by FTCM. Both enzymes preparations released cellobiose and glucose from pulps, with the cocktail from Trichoderma being the most efficient. Enzymatic treatments were not as effective at converting chemical-thermomechanical pulps to simple sugars, regardless of wood type. FTCM revealed that amorphous cellulose was the primary target of either enzyme preparation, which resulted in a higher proportion of crystalline cellulose on the surface after enzymatic treatment. FTCM confirmed that enzymes from Aspergillus had little impact on exposed hemicelluloses, but that enzymes from the more aggressive Trichoderma cocktail reduced hemicelluloses at the surface.ConclusionsOverall, this study indicates that treatment with enzymes from Trichoderma is appropriate for generating crystalline cellulose at fiber surface. Applications such as nanocellulose or composites requiring chemical resistance would benefit from this enzymatic treatment. The milder enzyme mixture from Aspergillus allowed for removal of amorphous cellulose while preserving hemicelluloses at fiber surface, which makes this treatment appropriate for new paper products where surface chemical responsiveness is required.Electronic supplementary materialThe online version of this article (10.1186/s13068-017-0980-0) contains supplementary material, which is available to authorized users.

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Section: Resultssupporting

confidence: 86%

Predicting the most appropriate wood biomass for selected industrial applications: comparison of wood, pulping, and enzymatic treatments using fluorescent-tagged carbohydrate-binding modules

Bombeck

Khatri

Meddeb-Mouelhi

et al. 2017

Biotechnol Biofuels

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“…So the ideal, or even requisite, proportion of accessory enzymes should be identified in future studies. For example of the proportion of enzymes, an ideal ratio was reported for the saccharification of kraft pulp in our previous paper, in the case of four types of cellulose degrading enzymes from fungus . The number of the different types of accessory enzymes, however, makes it difficult to identify the ratio of accessory enzymes when using previous methods.…”

Section: Discussionmentioning

confidence: 96%

Efficient and Supplementary Enzyme Cocktail from Actinobacteria and Plant Biomass Induction

et al. 2018

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Actinobacteria plays a key role in the cycling of organic matter in soils. They secret biomass-degrading enzymes that allow it to produce the unique metabolites that originate in plant biomass. Although past studies have focused on these unique metabolites, a large-scale screening of Actinobacteria is yet to be reported to focus on their biomass-degrading ability. In the present study, a rapid and simple method is constructed for a large-scale screening, and the novel resources that form the plant biomass-degrading enzyme cocktail are identified from 850 isolates of Actinobacteria. As a result, Nonomuraea fastidiosa secretes a biomass degrading enzyme cocktail with the highest enzyme titer, although cellulase activities are lower than a commercially available enzyme. So the rich accessory enzymes are suggested to contribute to the high enzyme titer for a pretreated bagasse with a synergistic effect. Additionally, an optimized cultivation method of biomass induction caused to produce the improved enzyme cocktail indicated strong enzyme titers and a strong synergistic effect. Therefore, the novel enzyme cocktails are selected via the optimized method for large-scale screening, and then the enzyme cocktail can be improved via the optimized production with biomass-induction.

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“…One unit of EG activity was defined as the absorption at 590 nm of released blue dye in 1 h at 38 °C (U/mL). p -Nitrophenyl- β -lactopyranoside ( p NPL, Sigma Co. Ltd, St. Louis, MO, USA) was used for the measurement of CBHI activity as previously described [ 49 ]. One unit of CBHI activity was defined as the enzyme amount required for production of 1 μmoL p NP in 1 min at 50 °C (U/mL).…”

Section: Methodsmentioning

confidence: 99%

Combined cell-surface display- and secretion-based strategies for production of cellulosic ethanol with Saccharomyces cerevisiae

Liu

Inokuma

et al. 2015

Biotechnol Biofuels

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BackgroundEngineering Saccharomyces cerevisiae to produce heterologous cellulases is considered as a promising strategy for production of bioethanol from lignocellulose. The production of cellulase is usually pursued by one of the two strategies: displaying enzyme on the cell surface or secreting enzyme into the medium. However, to our knowledge, the combination of the two strategies in a yeast strain has not been employed.ResultsIn this study, heterologous endoglucanase (EG) and cellobiohydrolase I (CBHI) were produced in a β-glucosidase displaying S. cerevisiae strain using cell-surface display, secretion, or a combined strategy. Strains EG-D-CBHI-D and EG-S-CBHI-S (with both enzymes displayed on the cell surface or with both enzymes secreted to the surrounding medium) showed higher ethanol production (2.9 and 2.6 g/L from 10 g/L phosphoric acid swollen cellulose, respectively), than strains EG-D-CBHI-S and EG-S-CBHI-D (with EG displayed on cell surface and CBHI secreted, or vice versa). After 3-cycle repeated-batch fermentation, the cellulose degradation ability of strain EG-D-CBHI-D remained 60 % of the 1st batch, at a level that was 1.7-fold higher than that of strain EG-S-CBHI-S.ConclusionsThis work demonstrated that placing EG and CBHI in the same space (on the cell surface or in the medium) was favorable for amorphous cellulose-based ethanol fermentation. In addition, the cellulolytic yeast strain that produced enzymes by the cell-surface display strategy performed better in cell-recycle batch fermentation compared to strains producing enzymes via the secretion strategy.Electronic supplementary materialThe online version of this article (doi:10.1186/s13068-015-0344-6) contains supplementary material, which is available to authorized users.

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Effective saccharification of kraft pulp by using a cellulase cocktail prepared from genetically engineered Aspergillus oryzae

Abstract: Akihiko Kondo (2015) Effective saccharification of kraft pulp by using a cellulase cocktail prepared from genetically engineered Aspergillus oryzae, Bioscience, Biotechnology, and Biochemistry, 79:6,

Cited by 9 publications

References 13 publications

Predicting the most appropriate wood biomass for selected industrial applications: comparison of wood, pulping, and enzymatic treatments using fluorescent-tagged carbohydrate-binding modules

Predicting the most appropriate wood biomass for selected industrial applications: comparison of wood, pulping, and enzymatic treatments using fluorescent-tagged carbohydrate-binding modules

Efficient and Supplementary Enzyme Cocktail from Actinobacteria and Plant Biomass Induction

Combined cell-surface display- and secretion-based strategies for production of cellulosic ethanol with Saccharomyces cerevisiae

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