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      Mild chemical pretreatments are sufficient for bioethanol production in transgenic rice straws overproducing glucosidase

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          Abstract

          A cost-effective biomass conversion technology for high bioethanol production under mild chemical pretreatment in the transgenic rice straw.

          Abstract

          Rice is a major food crop containing large amounts of lignocellulose residues usable for biofuels. In this study, we collected transgenic rice plants that over-produced Trichoderma reesei β-1,4- d-glucosidase (BGL I) into the cell walls in the mature straws. Without any pretreatment, the transgenic rice straws showed a consistently higher biomass enzymatic saccharification than the wild-type (WT) cultivar, in particular when 1% Tween-80 or 0.5% PEG-4000 was co-supplied into the enzymatic hydrolysis. Notably, under mild alkali pretreatment (1% NaOH at 50 °C for 2 h), the desirable transgenic line exhibited complete biomass enzymatic hydrolysis, resulting in the highest bioethanol yield of 21% (% dry matter) when compared with the rice and other bioenergy crops subjected to stronger pretreatment conditions reported in previous studies. Meanwhile, despite relatively low hexose yields obtained under 1% H 2SO 4 pretreatment, the transgenic rice straw also showed high bioethanol production at 18% due to an almost complete sugar–ethanol conversion rate. Chemical analyses indicated that the transgenic rice straw had significantly increased biomass porosity and reduced cellulose features (CrI, DP), which contributed to the largely enhanced biomass enzymatic hydrolysis. In addition, the raised arabinose level in hemicellulose and the lignin H-monomer proportion may also positively affect the biomass enzymatic saccharification in the transgenic rice straw. Hence, this study demonstrated a cost-effective and green lignocellulose conversion technology for high bioethanol production in the transgenic rice straw. It also provided a strategy for the potential genetic modification of plant cell walls in bioenergy crops.

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          Most cited references57

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          Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA.

          A large number of morphologically normal, fertile, transgenic rice plants were obtained by co-cultivation of rice tissues with Agrobacterium tumefaciens. The efficiency of transformation was similar to that obtained by the methods used routinely for transformation of dicotyledons with the bacterium. Stable integration, expression and inheritance of transgenes were demonstrated by molecular and genetic analysis of transformants in the R0, R1 and R2 generations. Sequence analysis revealed that the boundaries of the T-DNA in transgenic rice plants were essentially identical to those in transgenic dicotyledons. Calli induced from scutella were very good starting materials. A strain of A. tumefaciens that carried a so-called 'super-binary' vector gave especially high frequencies of transformation of various cultivars of japonica rice that included Koshihikari, which normally shows poor responses in tissue culture.
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            Cellulose crystallinity index: measurement techniques and their impact on interpreting cellulase performance

            Although measurements of crystallinity index (CI) have a long history, it has been found that CI varies significantly depending on the choice of measurement method. In this study, four different techniques incorporating X-ray diffraction and solid-state 13C nuclear magnetic resonance (NMR) were compared using eight different cellulose preparations. We found that the simplest method, which is also the most widely used, and which involves measurement of just two heights in the X-ray diffractogram, produced significantly higher crystallinity values than did the other methods. Data in the literature for the cellulose preparation used (Avicel PH-101) support this observation. We believe that the alternative X-ray diffraction (XRD) and NMR methods presented here, which consider the contributions from amorphous and crystalline cellulose to the entire XRD and NMR spectra, provide a more accurate measure of the crystallinity of cellulose. Although celluloses having a high amorphous content are usually more easily digested by enzymes, it is unclear, based on studies published in the literature, whether CI actually provides a clear indication of the digestibility of a cellulose sample. Cellulose accessibility should be affected by crystallinity, but is also likely to be affected by several other parameters, such as lignin/hemicellulose contents and distribution, porosity, and particle size. Given the methodological dependency of cellulose CI values and the complex nature of cellulase interactions with amorphous and crystalline celluloses, we caution against trying to correlate relatively small changes in CI with changes in cellulose digestibility. In addition, the prediction of cellulase performance based on low levels of cellulose conversion may not include sufficient digestion of the crystalline component to be meaningful.
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              Pretreatments to enhance the digestibility of lignocellulosic biomass.

              Lignocellulosic biomass represents a rather unused source for biogas and ethanol production. Many factors, like lignin content, crystallinity of cellulose, and particle size, limit the digestibility of the hemicellulose and cellulose present in the lignocellulosic biomass. Pretreatments have as a goal to improve the digestibility of the lignocellulosic biomass. Each pretreatment has its own effect(s) on the cellulose, hemicellulose and lignin; the three main components of lignocellulosic biomass. This paper reviews the different effect(s) of several pretreatments on the three main parts of the lignocellulosic biomass to improve its digestibility. Steam pretreatment, lime pretreatment, liquid hot water pretreatments and ammonia based pretreatments are concluded to be pretreatments with high potentials. The main effects are dissolving hemicellulose and alteration of lignin structure, providing an improved accessibility of the cellulose for hydrolytic enzymes.
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                Author and article information

                Journal
                GRCHFJ
                Green Chemistry
                Green Chem.
                Royal Society of Chemistry (RSC)
                1463-9262
                1463-9270
                2018
                2018
                : 20
                : 9
                : 2047-2056
                Affiliations
                [1 ]Biomass and Bioenergy Research Centre
                [2 ]Huazhong Agricultural University
                [3 ]Wuhan
                [4 ]China
                [5 ]College of Plant Science and Technology
                [6 ]College of Life Science and Technology
                Article
                10.1039/C8GC00694F
                87627c15-60f6-49d0-bf8c-c1ff93e9f725
                © 2018

                http://rsc.li/journals-terms-of-use

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