Institute for Molecular Biotechnology (Biology VII), RWTH Aachen University, Worringerweg 1, 52074, Aachen, Germany.
Biotechnol Biofuels. 2012 Aug 28;5(1):63. doi: 10.1186/1754-6834-5-63.
The enzymatic conversion of lignocellulosic plant biomass into fermentable sugars is a crucial step in the sustainable and environmentally friendly production of biofuels. However, a major drawback of enzymes from mesophilic sources is their suboptimal activity under established pretreatment conditions, e.g. high temperatures, extreme pH values and high salt concentrations. Enzymes from extremophiles are better adapted to these conditions and could be produced by heterologous expression in microbes, or even directly in the plant biomass.
Here we show that a cellulase gene (sso1354) isolated from the hyperthermophilic archaeon Sulfolobus solfataricus can be expressed in plants, and that the recombinant enzyme is biologically active and exhibits the same properties as the wild type form. Since the enzyme is inactive under normal plant growth conditions, this potentially allows its expression in plants without negative effects on growth and development, and subsequent heat-inducible activation. Furthermore we demonstrate that the recombinant enzyme acts in high concentrations of ionic liquids and can therefore degrade α-cellulose or even complex cell wall preparations under those pretreatment conditions.
The hyperthermophilic endoglucanase SSO1354 with its unique features is an excellent tool for advanced biomass conversion. Here we demonstrate its expression in planta and the possibility for post harvest activation. Moreover the enzyme is suitable for combined pretreatment and hydrolysis applications.
木质纤维素植物生物质转化为可发酵糖是生物燃料可持续和环保生产的关键步骤。然而,来自嗜温源的酶的一个主要缺点是它们在既定预处理条件下(例如高温、极端 pH 值和高盐浓度)的活性不佳。来自极端微生物的酶更好地适应这些条件,可以通过异源表达在微生物中,甚至可以直接在植物生物质中产生。
在这里,我们表明,从嗜热古菌 Sulfolobus solfataricus 中分离的纤维素酶基因(sso1354)可以在植物中表达,并且重组酶具有生物活性并且表现出与野生型相同的性质。由于该酶在正常植物生长条件下失活,因此这可能允许其在植物中表达而不会对生长和发育产生负面影响,并且随后可以进行热诱导激活。此外,我们证明该重组酶在高浓度离子液体中起作用,因此可以在这些预处理条件下降解α-纤维素甚至复杂的细胞壁制剂。
具有独特特征的嗜热内切葡聚糖酶 SSO1354 是先进生物质转化的绝佳工具。在这里,我们证明了它在植物中的表达和收获后激活的可能性。此外,该酶适用于组合预处理和水解应用。