Pablo Alvira

Pretreatment technologies for an efficient bioethanol production process based on enzymatic hydrolysis: A review

Biofuel produced from lignocellulosic materials, so-called second generation bioethanol shows energetic, economic and environmental advantages in comparison to bioethanol from starch or sugar. However, physical and chemical barriers caused by the close association of the main components of lignocellulosic biomass, hinder the hydrolysis of cellulose and hemicellulose to fermentable sugars. The main goal of pretreatment is to increase the enzyme accessibility improving digestibility of cellulose. Each pretreatment has a specific effect on the cellulose, hemicellulose and lignin fraction thus, different pretreatment methods and conditions should be chosen according to the process configuration selected for the subsequent hydrolysis and fermentation steps. This paper reviews the most interesting technologies for ethanol production from lignocellulose and it points out several key properties that should be targeted for low-cost and advanced pretreatment processes.

Effect of endoxylanase and α-L-arabinofuranosidase supplementation on the enzymatic hydrolysis of steam exploded wheat straw.

The cost and hydrolytic efficiency of enzymes are major factors that restrict the commercialization of the bioethanol production process from lignocellulosic biomass. Hemicellulases and other accessory enzymes are becoming crucial to increase enzymatic hydrolysis (EH) yields at low cellulase dosages. The aim of this work was to evaluate the effect of two recombinant hemicellulolytic enzymes on the EH of steam pretreated wheat straw. Pretreatments at two severity conditions were performed and the whole slurry obtained after steam explosion pretreatment was employed as substrate. An endoxylanase (Xln C) from Aspergillus nidulans and an α-L-arabinofuranosidase (AF) from Aspergillus niger, have been applied in combination with cellulase enzymes. A degree of synergism of 29.5% and increases up to 10% in the EH yields were obtained, showing the potential of accessory activities to improve the EH step and make the whole process more effective.

A review of biological delignification and detoxification methods for lignocellulosic bioethanol production

Abstract Future biorefineries will integrate biomass conversion processes to produce fuels, power, heat and value-added chemicals. Due to its low price and wide distribution, lignocellulosic biomass is expected to play an important role toward this goal. Regarding renewable biofuel production, bioethanol from lignocellulosic feedstocks is considered the most feasible option for fossil fuels replacement since these raw materials do not compete with food or feed crops. In the overall process, lignin, the natural barrier of the lignocellulosic biomass, represents an important limiting factor in biomass digestibility. In order to reduce the recalcitrant structure of lignocellulose, biological pretreatments have been promoted as sustainable and environmentally friendly alternatives to traditional physico-chemical technologies, which are expensive and pollute the environment. These approaches include the use of diverse white-rot fungi and/or ligninolytic enzymes, which disrupt lignin polymers and facilitate the bioconversion of the sugar fraction into ethanol. As there is still no suitable biological pretreatment technology ready to scale up in an industrial context, white-rot fungi and/or ligninolytic enzymes have also been proposed to overcome, in a separated or in situ biodetoxification step, the effect of the inhibitors produced by non-biological pretreatments. The present work reviews the latest studies regarding the application of different microorganisms or enzymes as useful and environmentally friendly delignification and detoxification technologies for lignocellulosic biofuel production. This review also points out the main challenges and possible ways to make these technologies a reality for the bioethanol industry.

Improving the fermentation performance of saccharomyces cerevisiae by laccase during ethanol production from steam-exploded wheat straw at high-substrate loadings

Operating the saccharification and fermentation processes at high‐substrate loadings is a key factor for making ethanol production from lignocellulosic biomass economically viable. However, increasing the substrate loading presents some disadvantages, including a higher concentration of inhibitors (furan derivatives, weak acids, and phenolic compounds) in the media, which negatively affect the fermentation performance. One strategy to eliminate soluble inhibitors is filtering and washing the pretreated material. In this study, it was observed that even if the material was previously washed, inhibitory compounds were released during the enzymatic hydrolysis step. Laccase enzymatic treatment was evaluated as a method to reduce these inhibitory effects. The laccase efficiency was analyzed in a presaccharification and simultaneous saccharification and fermentation process at high‐substrate loadings. Water‐insoluble solids fraction from steam‐exploded wheat straw was used as substrate and Saccharomyces cerevisiae as fermenting microorganism. Laccase supplementation reduced strongly the phenolic content in the media, without affecting weak acids and furan derivatives. This strategy resulted in an improved yeast performance during simultaneous saccharification and fermentation process, increasing significantly ethanol productivity.

Strategies of xylanase supplementation for an efficient saccharification and cofermentation process from pretreated wheat straw

Ethanol production from lignocellulosic raw materials includes a pretreatment step before enzymatic hydrolysis (EH). Pretreated substrates contain complex hemicelluloses in the solid fraction that can protect the cellulose from enzymatic attack. In addition, soluble xylooligomers are contained in the pretreated materials and may have an inhibitory effect on cellulase activity. In this context, several approaches for xylanase supplementation have been studied to increase EH yields. In this study, the whole slurry obtained after steam explosion pretreatment of wheat straw has been used as substrate. EH experiments were performed using commercial cellulase preparations supplemented with an endoxylanase (XlnC) from Aspergillus nidulans. Among different strategies of XlnC supplementation, the 24‐h xylanase treatment before cellulase addition yielded an increase of 40.1 and 10.1% in glucose and xylose production, respectively. Different XlnC addition strategies were integrated in a simultaneous saccharification and cofermentation process (SSCF) using the xylose fermenting strain Saccharomyces cerevisiae F12. Ethanol production in SSCF was 28.4% higher when comparing to a simultaneous saccharification and fermentation process.

Progress on Enzymatic Saccharification Technologies for Biofuels Production

A key issue for the biorefineries is the cost-effective conversion of carbohydrates contained in lignocellulosic biomass into fermentable sugars, which will provide a viable route to biofuels and bioproducts. Many different raw materials, conversion methods, and process configurations have been studied for the generation of sugars from lignocellulosic biomass. Most of the schemes for lignocellulosic biomass conversion include a pretreatment step to increase digestibility of the substrates and an enzymatic hydrolysis process, which is a crucial step and determines the overall process efficiency. Due to lignocellulose complex structure, different enzymes are involved in the degradation of the substrates and appropriate combinations of different activities are required for complete hydrolysis. This chapter reviews novel advances in enzymatic hydrolysis technologies for lignocellulose conversion, with special focus on the necessity of optimized enzyme mixtures using accessory activities, and the advantages of operating at high initial substrate concentrations.

Steam Explosion for Wheat Straw Pretreatment for Sugars Production

Abstract Development of biofuels such as lignocellulosic ethanol represents a sustainable alternative in the transport sector. Wheat straw is a promising feedstock for bioethanol production in Europe due to its large production and high carbohydrates content. In a process to produce cellulosic ethanol, previous to the enzymatic hydrolysis to obtain fermentable sugars and the subsequent fermentation, a pretreatment step to break down the recalcitrance of lignocellulose fiber is essential. In this work, a range of steam explosion pretreatment conditions were evaluated according to different parameters: sugars recovery, degradation products generation, and enzymatic hydrolysis yields. Moreover, the enzymatic hydrolysis process was also studied at high substrate loadings, since operating at high solids loading is crucial for large scale development of ethanol production. Pretreatment at 200°C - 10 min resulted in higher enzymatic hydrolysis yield (91.7%) and overall glucose yields (35.4 g glucose/100 g wheat straw) but also higher production of toxic compound. In turn, the characteristics of the pretreated wheat straw at lower severity (Log R0=3.65) correspond to 190°C and 10 min, with minimal sugars degradation and toxics formation indicated a great potential for maximizing total sugars production by using optimal enzyme combinations including accessory enzymes in the enzymatic hydrolysis step.

Cellulase production using different streams of wheat grain- and wheat straw-based ethanol processes

Pretreatment is a necessary step in the biomass-to-ethanol conversion process. The side stream of the pretreatment step is the liquid fraction, also referred to as the hydrolyzate, which arises after the separation of the pretreated solid and is composed of valuable carbohydrates along with compounds that are potentially toxic to microbes (mainly furfural, acetic acid, and formic acid). The aim of our study was to utilize the liquid fraction from steam-exploded wheat straw as a carbon source for cellulase production by Trichodermareesei RUT C30. Results showed that without detoxification, the fungus failed to utilize any dilution of the hydrolyzate; however, after a two-step detoxification process, it was able to grow on a fourfold dilution of the treated liquid fraction. Supplementation of the fourfold-diluted, treated liquid fraction with washed pretreated wheat straw or ground wheat grain led to enhanced cellulase (filter paper) activity. Produced enzymes were tested in hydrolysis of washed pretreated wheat straw. Supplementation with ground wheat grain provided a more efficient enzyme mixture for the hydrolysis by means of the near-doubled β-glucosidase activity obtained.