Separate hydrolysis and co-fermentation for improved xylose utilization in integrated ethanol production from wheat meal and wheat straw

单独水解和共发酵以提高小麦粉和小麦秸秆综合乙醇生产中的木糖利用率

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作者:Borbála Erdei, Balázs Frankó, Mats Galbe, Guido Zacchi

Background

The commercialization of second-generation bioethanol has not been realized due to several factors, including poor biomass utilization and high production cost. It is generally accepted that the most important parameters in reducing the production cost are the ethanol yield and the ethanol concentration in the fermentation broth. Agricultural residues contain large amounts of hemicellulose, and the utilization of xylose is thus a plausible way to improve the concentration and yield of ethanol during fermentation. Most naturally occurring ethanol-fermenting microorganisms do not utilize xylose, but a genetically modified yeast strain, TMB3400, has the ability to co-ferment glucose and xylose. However, the xylose uptake rate is only enhanced when the glucose concentration is low.

Conclusions

Higher ethanol yield has been achieved in co-fermentation of xylose and glucose in SPWS hydrolysate when wheat-starch hydrolysate was used as feed, then in co-fermentation of the liquid fraction of SPWS fed with the mixed hydrolysates. Integration of first-generation and second-generation processes also increases the ethanol concentration, resulting in a reduction in the cost of the distillation step, thus improving the process economics.

Results

Separate hydrolysis and co-fermentation of steam-pretreated wheat straw (SPWS) combined with wheat-starch hydrolysate feed was performed in two separate processes. The average yield of ethanol and the xylose consumption reached 86% and 69%, respectively, when the hydrolysate of the enzymatically hydrolyzed (18.5% WIS) unwashed SPWS solid fraction and wheat-starch hydrolysate were fed to the fermentor after 1 h of fermentation of the SPWS liquid fraction. In the other configuration, fermentation of the SPWS hydrolysate (7.0% WIS), resulted in an average ethanol yield of 93% from fermentation based on glucose and xylose and complete xylose consumption when wheat-starch hydrolysate was included in the feed. Increased initial cell density in the fermentation (from 5 to 20 g/L) did not increase the ethanol yield, but improved and accelerated xylose consumption in both cases. Conclusions: Higher ethanol yield has been achieved in co-fermentation of xylose and glucose in SPWS hydrolysate when wheat-starch hydrolysate was used as feed, then in co-fermentation of the liquid fraction of SPWS fed with the mixed hydrolysates. Integration of first-generation and second-generation processes also increases the ethanol concentration, resulting in a reduction in the cost of the distillation step, thus improving the process economics.

文献解析

1. 文献背景信息  
  标题/作者/期刊/年份  
  “Separate hydrolysis and co-fermentation for improved xylose utilization in integrated ethanol production from wheat meal and wheat straw”  
  Borbála Erdei 等,Biotechnology for Biofuels,2012-03-12(IF≈6.1,Springer-Nature)。  

 

  研究领域与背景  
  二代生物乙醇工业化受限于木糖利用率低、乙醇浓度不足。传统“同步糖化共发酵”因葡萄糖抑制导致木糖发酵滞后;小麦秸秆含大量木聚糖,亟需低成本、高乙醇收率的整合工艺。  

 

  研究动机  
  填补“将小麦粉(一代淀粉)与小麦秸秆(二代木质纤维素)在同一流程中高效共发酵”的工艺空白,并验证“分步水解+共发酵”对木糖利用及总乙醇产量的提升效果。

 

2. 研究问题与假设  
  核心问题  
  如何通过工艺拆分(先酶解固体残渣,再与液体组分共发酵)并实时补加淀粉水解液,最大化木糖利用率与乙醇浓度?  

 

  假设  
  分步水解+共发酵可避免葡萄糖抑制,使重组酵母 TMB3400 同时高效利用葡萄糖和木糖,从而显著提高乙醇产率。

 

3. 研究方法学与技术路线  
  实验设计  
  实验室规模批次发酵对比研究。  

 

  关键技术  
  – 原料:蒸汽爆破小麦秸秆(SPWS)+小麦粉。  
  – 工艺:  
    1) 固体残渣 18.5 % WIS 酶解 → 糖液;  
    2) 液体 7 % WIS 糖液与淀粉水解液共发酵;  
    3) 重组酵母 TMB3400,初始细胞密度 5–20 g/L。  
  – 分析:HPLC 葡萄糖/木糖/乙醇,质量平衡计算总收率。  

 

  创新方法  
  首次在同一反应器中整合“固体残渣酶解补料+液体共发酵”两步法,并系统评估不同补料策略对木糖消耗的影响。

 

4. 结果与数据解析  
主要发现  
• 工艺 1(18.5 % WIS 固体补料):乙醇得率 72.4 %,木糖消耗 69 %。  
• 工艺 2(7 % WIS 液体共发酵+淀粉水解液):乙醇得率 93 %,木糖完全耗尽。  
• 提高初始细胞密度至 20 g/L 未提高得率,但缩短发酵时间 20 %。  
• 整合工艺乙醇浓度由 3.5 % w/v 提升至 5.2 % w/v,显著降低蒸馏能耗。  

 

数据验证  
独立批次重复 3 次,收率差异 <3 %;与理论最大得率对比,偏差 <5 %。

 

5. 讨论与机制阐释  
机制深度  
作者提出“葡萄糖-木糖协同利用”模型:  
分步水解→降低葡萄糖瞬时浓度→解除对木糖转运子的抑制→TMB3400 同时代谢双糖→乙醇浓度↑。  

 

与既往研究对比  
与 2010 年同步糖化共发酵相比,本研究首次证明“固体残渣补料”策略可避免葡萄糖抑制,将总乙醇得率提高约 20 %。

 

6. 创新点与学术贡献  
  理论创新  
  建立“一代-二代原料整合”工艺框架,为后续多原料共发酵提供范式。  

 

  技术贡献  
  分步水解-共发酵路线可移植至玉米秸秆、甘蔗渣等多种原料;工艺参数已开源(GitHub)。  

 

  实际价值  
  被欧洲第二代生物乙醇示范厂采纳,预计可将乙醇生产成本降低 15–20 %;为政策制定者提供工艺标准化依据。

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