How recombinant swollenin from Kluyveromyces lactis affects cellulosic substrates and accelerates their hydrolysis.
How recombinant swollenin from Kluyveromyces lactis affects cellulosic substrates and accelerates their hydrolysis.
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乳酸乳腺菌的重组肿瘤蛋白如何影响纤维素底物并加速其水解。
DOI：
10.1186/1754-6834-4-33
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发表时间：
2011-09-23
影响因子：
6.3
通讯作者：
Büchs J
中科院分区：
文献类型：
--
作者：
Jäger G;Girfoglio M;Dollo F;Rinaldi R;Bongard H;Commandeur U;Fischer R;Spiess AC;Büchs J
为了产生生物燃料，不溶性纤维素底物被预处理，随后用纤维素酶水解。以安全和环境友好的方式预处理纤维素的一种方法是在温和的条件下应用非水解蛋白质，例如膨胀素-由真菌里氏木霉以低产量天然产生。为了获得足够的膨胀素用于工业应用，本研究的第一个目的是提出一种新的生产重组膨胀素的方法。主要目的是显示膨胀素如何定量地影响纤维素底物的相关物理性质，以及它如何影响随后的水解。在乳酸克鲁维酵母中表达后，纯化所得膨胀素。首次对重组膨胀素在纤维素上的吸附参数进行了定量，并与T. reesei。然后用膨胀素预处理四种不同的不溶性纤维素底物。首先，可以通过宏观评价和显微镜定性地显示膨胀素引起较大纤维素聚集体的解聚以及纤维素微纤丝的分散（无定形）。之后，膨胀素对纤维素颗粒大小，最大纤维素酶吸附和纤维素结晶度的影响进行了定量。用膨胀素预处理导致纤维素底物的粒度以及它们的结晶度显著降低，从而显著增加了纤维素酶在这些底物上的最大吸附。随后，用纤维素酶水解预处理的纤维素底物。在这里，用膨胀素预处理纤维素底物，即使在非饱和浓度下，也显著加速水解。通过将纤维素底物的粒度和结晶度与初始水解速率相关联，可以表明膨胀素诱导的粒度和结晶度的降低导致高的纤维素水解速率。重组膨胀素可以很容易地用鲁棒酵母K.乳酸菌。此外，膨胀素诱导纤维素团聚体的解聚以及无定形（脱钙）。这项研究第一次量化并详细阐述了膨胀素如何影响不同的纤维素底物及其水解。
In order to generate biofuels, insoluble cellulosic substrates are pretreated and subsequently hydrolyzed with cellulases. One way to pretreat cellulose in a safe and environmentally friendly manner is to apply, under mild conditions, non-hydrolyzing proteins such as swollenin - naturally produced in low yields by the fungus Trichoderma reesei. To yield sufficient swollenin for industrial applications, the first aim of this study is to present a new way of producing recombinant swollenin. The main objective is to show how swollenin quantitatively affects relevant physical properties of cellulosic substrates and how it affects subsequent hydrolysis. After expression in the yeast Kluyveromyces lactis, the resulting swollenin was purified. The adsorption parameters of the recombinant swollenin onto cellulose were quantified for the first time and were comparable to those of individual cellulases from T. reesei. Four different insoluble cellulosic substrates were then pretreated with swollenin. At first, it could be qualitatively shown by macroscopic evaluation and microscopy that swollenin caused deagglomeration of bigger cellulose agglomerates as well as dispersion of cellulose microfibrils (amorphogenesis). Afterwards, the effects of swollenin on cellulose particle size, maximum cellulase adsorption and cellulose crystallinity were quantified. The pretreatment with swollenin resulted in a significant decrease in particle size of the cellulosic substrates as well as in their crystallinity, thereby substantially increasing maximum cellulase adsorption onto these substrates. Subsequently, the pretreated cellulosic substrates were hydrolyzed with cellulases. Here, pretreatment of cellulosic substrates with swollenin, even in non-saturating concentrations, significantly accelerated the hydrolysis. By correlating particle size and crystallinity of the cellulosic substrates with initial hydrolysis rates, it could be shown that the swollenin-induced reduction in particle size and crystallinity resulted in high cellulose hydrolysis rates. Recombinant swollenin can be easily produced with the robust yeast K. lactis. Moreover, swollenin induces deagglomeration of cellulose agglomerates as well as amorphogenesis (decrystallization). For the first time, this study quantifies and elucidates in detail how swollenin affects different cellulosic substrates and their hydrolysis.
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影响因子：
3.8
作者：
BELDMAN, G;VORAGEN, AGJ;PILNIK, W
通讯作者：
PILNIK, W
影响因子：
3.8
作者：
Gupta, Rajesh;Lee, Y. Y.
通讯作者：
Lee, Y. Y.
影响因子：
5.4
作者：
Hall, Melanie;Bansal, Prabuddha;Bommarius, Andreas S.
通讯作者：
Bommarius, Andreas S.
影响因子：
3.4
作者：
Cao, Y;Tan, HM
通讯作者：
Tan, HM
纤维素的获取限制了酶水解的效率：无定形发生的作用。
影响因子：
6.3
作者：
Arantes V;Saddler JN
通讯作者：
Saddler JN