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    • 2. 发明申请
    • CELLULOSIC PROCESSING TRAIT DEVELOPMENT USING A THERMOREGULATED, INTEIN-MODIFIED XYLANASE
    • 使用加热改性的XYLANASE进行的细胞处理过程开发
    • US20130203125A1
    • 2013-08-08
    • US13818928
    • 2011-08-23
    • Binzhang ShenJames ApgarOleg BougriR. Michael Raab
    • Binzhang ShenJames ApgarOleg BougriR. Michael Raab
    • C12N15/82C12N9/24C12P19/14
    • C12N15/8243C12N9/248C12N9/2482C12N15/8216C12N15/8246C12P19/14C12Y302/01008
    • In planta consolidated bioprocessing has the advantages of decreasing biomass pretreatment costs, utilizing excess plant protein production capacity for enzyme production, and decreasing mass transfer resistance of enzyme diffusion to its substrate. However, in planta expression of cell wall degrading (CWD) enzymes often leads to detrimental plant phenotypes that impact crop yield. To provide in planta CWD enzyme activity without any adverse phenotype, a thermostable xylanase, XynB (EC 3.2.1.8), was engineered with a thermoregulated intein, Tth-HB27 DnaE-1 (Tth intein), that controls its hydrolytic activity through conditional intein splicing. Maize plants expressing the heat inducible Tth intein-modified XynB developed normally, yet possessed enhanced post harvest glucose production from dried corn stover. Expression of CWD enzymes as dormant, intein-modified proteins that can be activated by heat treatment after harvest provides the basis for developing a novel cellulosic processing trait in plants.
    • 在植物中,综合生物处理具有降低生物量预处理成本,利用植物蛋白生产过剩的酶生产能力,降低酶扩散对其底物的传质阻力的优点。 然而,在植物表达的细胞壁降解(CWD)酶通常导致有害的植物表型影响作物产量。 为了在没有任何不良表型的情况下提供植物CWD酶活性,用热调节的内含肽Tth-HB27DnaE-1(第T个内含肽)设计热稳定的木聚糖酶XynB(EC 3.2.1.8),其通过条件内含肽来控制其水解活性 拼接。 表达热诱导性Tth内蛋白修饰的XynB的玉米植物正常发育,但是从干燥的玉米秸秆中获得增强的后收获葡萄糖产量。 CWD酶作为休眠后的内含肽修饰蛋白质的表达,可以在收获后通过热处理活化,为开发植物中新型纤维素加工性状提供依据。