Visualising recalcitrance by colocalisation of cellulase, lignin and cellulose in pretreated pine biomass using fluorescence microscopy.
Visualising recalcitrance by colocalisation of cellulase, lignin and cellulose in pretreated pine biomass using fluorescence microscopy.
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DOI：
10.1038/srep44386
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发表时间：
2017-03-10
影响因子：
4.6
通讯作者：
Vaidya A
中科院分区：
文献类型：
--
作者：
Donaldson L;Vaidya A
绘制结合的纤维素酶的位置，可为生物燃料应用中预处理木质生物质中易受作用和难降解位点的微观分布提供信息。利用共聚焦显微镜对荧光标记的纤维素酶混合物与蒸汽爆破松木（SEW）的相互作用进行了定量分析。通过使用皮尔逊相关性测量它们的共定位，相对于木质素（自发荧光）和纤维素（刚果红染色）对Dylight标记的纤维素酶的空间分布进行了定量。与富含木质素的中层相比，富含纤维素的次生细胞壁中的相关性更强，但在单个生物质颗粒之间存在显著差异。在校正木质素和纤维素之间的相关性后，预处理生物质中纤维素的分布占酶分布变化的30%。共定位分析首次能够根据纤维素和木质素的组织化学来量化易受作用和难降解位点的空间分布。这项研究将有助于理解预处理在难降解软木生物质酶水解中的作用。
Mapping the location of bound cellulase enzymes provides information on the micro-scale distribution of amenable and recalcitrant sites in pretreated woody biomass for biofuel applications. The interaction of a fluorescently labelled cellulase enzyme cocktail with steam-exploded pine (SEW) was quantified using confocal microscopy. The spatial distribution of Dylight labelled cellulase was quantified relative to lignin (autofluorescence) and cellulose (Congo red staining) by measuring their colocalisation using Pearson correlations. Correlations were greater in cellulose-rich secondary cell walls compared to lignin-rich middle lamella but with significant variations among individual biomass particles. The distribution of cellulose in the pretreated biomass accounted for 30% of the variation in the distribution of enzyme after correcting for the correlation between lignin and cellulose. For the first time, colocalisation analysis was able to quantify the spatial distribution of amenable and recalcitrant sites in relation to the histochemistry of cellulose and lignin. This study will contribute to understanding the role of pretreatment in enzymatic hydrolysis of recalcitrant softwood biomass.