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提高 Spathaspora passalidarum 对甘蔗渣水解液的耐受性,以从木糖生产乙醇。

Enhanced Tolerance of Spathaspora passalidarum to Sugarcane Bagasse Hydrolysate for Ethanol Production from Xylose.

机构信息

Embrapa Agroenergia, Parque Estação Biológica - PqEB s/no, W3 Norte (final), Brasília, DF, 70770-901, Brazil.

CIETI, School of Engineering (ISEP), Polytechnic Institute of Porto (P. Porto), R. Dr. António Bernardino de Almeida, 431, 4249-015, Porto, Portugal.

出版信息

Appl Biochem Biotechnol. 2021 Jul;193(7):2182-2197. doi: 10.1007/s12010-021-03544-6. Epub 2021 Mar 8.

Abstract

During the pretreatment and hydrolysis of lignocellulosic biomass to obtain a hydrolysate rich in fermentable sugars, furaldehydes (furfural and hydroxymethylfurfural), phenolic compounds, and organic acids are formed and released. These compounds inhibit yeast metabolism, reducing fermentation yields and productivity. This study initially confirmed the ability of Spathaspora passalidarum to ferment xylose and demonstrated its sensibility to the inhibitors present in the hemicellulosic sugarcane bagasse hydrolysate. Then, an adaptive laboratory evolution, with progressive increments of hydrolysate concentration, was employed to select a strain more resistant to hydrolysate inhibitors. Afterward, a central composite design was performed to maximize ethanol production using hydrolysate as substrate. At optimized conditions (initial cell concentration of 30 g/L), S. passalidarum was able to produce 19.4 g/L of ethanol with productivity, yield, and xylose consumption rate of 0.8 g/L.h and 0.4 g/g, respectively, in a sugarcane bagasse hemicellulosic hydrolysate. A kinetic model was developed to describe the inhibition of fermentation by substrate and product. The values obtained for substrate saturation and inhibition constant were K = 120.4 g/L and K = 1293.4 g/L. Ethanol concentration that stops cell growth was 30.1 g/L. There was an agreement between simulated and experimental results, with a residual standard deviation lower than 6%.

摘要

在预处理和水解木质纤维素生物质以获得富含可发酵糖的水解物的过程中,会形成并释放糠醛(糠醛和羟甲基糠醛)、酚类化合物和有机酸。这些化合物抑制酵母代谢,降低发酵产率和生产力。本研究最初证实了 Spathaspora passalidarum 发酵木糖的能力,并证明了其对存在于半纤维素甘蔗渣水解物中的抑制剂的敏感性。然后,采用渐进增加水解物浓度的适应性实验室进化来选择对水解物抑制剂更具抗性的菌株。随后,采用中心复合设计,使用水解物作为底物来最大化乙醇的产量。在优化条件下(初始细胞浓度为 30 g/L),S. passalidarum 能够在甘蔗渣半纤维素水解物中产生 19.4 g/L 的乙醇,其生产率、产率和木糖消耗率分别为 0.8 g/L.h 和 0.4 g/g。建立了一个动力学模型来描述底物和产物对发酵的抑制作用。获得的底物饱和和抑制常数的值分别为 K = 120.4 g/L 和 K = 1293.4 g/L。停止细胞生长的乙醇浓度为 30.1 g/L。模拟结果与实验结果吻合良好,残差标准偏差低于 6%。

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