RESUMEN

Lignocellulosic biomass has emerged as a promising environmental resource. Enzyme catalysis, as one of the most environmentally friendly and efficient tools among various treatments, is used for the conversion of biomass into chemicals and fuels. Cellulase is a complex enzyme composed of ß-glucosidase (BGL), endo-ß-1,4-glucanase (EG), and exo-ß-1,4-glucanase (CBH), which synergistically hydrolyzes cellulose into monosaccharides. BGL, which further deconstructs cellobiose and short-chain cellooligosaccharides obtained by EG and CBH catalysis into glucose, is the most sensitive component of the synergistic enzyme system constituted by the three enzymes and is highly susceptible to inactivation by external conditions, becoming the rate-limiting component in biomass conversion. This paper firstly introduces the source and catalytic mechanism of BGL used in the process of biomass resource utilization. The focus is on the review of various factors affecting BGL activity during hydrolysis, including competitive adsorption of lignin, gas-liquid interface inactivation, thermal inactivation, and solvent effect. And the methods to improve BGL inactivation are proposed from two aspects-substrate initiation and enzyme initiation. In particular, the screening, modification, and alteration of the enzyme molecules themselves are discussed with emphasis. This review can provide novel ideas for studies of BGL inactivation mechanism, containment of inactivation, and activity enhancement. KEY POINTS: • Factors affecting ß-glucosidase inactivation are described. • Process intensification is presented in terms of substrate and enzyme. • Solvent selection, protein engineering, and immobilization remain topics of interest.

Asunto(s)

RESUMEN

Lignocellulose, a kind of biological resource widely existing in nature, which can be transformed into value-added biochemical products through saccharification, fermentation or chemical catalysis. Pretreatments are the necessary step to increase the accessibility and digestibility of lignocellulose. This paper comprehensively reviewed different pretreatment progress of lignocellulose in recent year, including mechanical/thermal, biological, inorganic solvent, organic solvent and unconventional physical-chemical pretreatments, focusing on quantifying the influence of pretreatments on subsequent biomass conversion. In addition, related pretreatment techniques such as genetic engineering, reactor configurations, downstream process and visualization technology of pretreatment were discussed. Finally, this review presented the challenge of lignocellulose pretreatment in the future.

Asunto(s)

RESUMEN

The intrinsic recalcitrance of lignocellulosic biomass makes it resistant to enzymatic hydrolysis. The electron-rich surface of the lignin and cellulose-alike structure of hemicellulose competitively absorb the cellulase. Thus, modifying the surface charge on biomass components to alter cellulase affinity is an urgent requisite. Developing charge tunable cellulase will alter substrate affinity. Also, charge-based immobilization generates controllable substrate affinity. Within immobilized cellulase involved in situ biomass saccharification, charge effects made a crucial contribution. In addition to affecting the interaction between immobilized cellulase and biomass, charge exerts an impact on cellulase to immobilize the materials, further investigation is essential. This study aims to review the charge effects on the cellulase affinity in biomass saccharification, strategies of charge tunable cellulase, and immobilized cellulase, thereby explaining the role of electrostatic interaction. In terms of electrostatic behavior, the pathways and plans to improve in situ biomass saccharification seem to be promising.

Asunto(s)

RESUMEN

Herbel-tolerant strains exhibit considerable environmental and commercial values not only due to their harmless treatment of herbal-extraction residues (HERs) but also because of their use in preparing high-quality cellulase cocktails. In this study, three typical HERs were evaluated for enzymatic in situ saccharification performance. A HERs-tolerance fungus, identified as Penicillium oxalicum G2, can grow in 1.5% (w/v) Radix isatidis residues (RIR), thereby exhibiting the highest FPase (2.2 U/mL), carboxymethyl cellulase (13.3 U/mL), and ß-glucosidase (4.6 U/mL) activities. The most effective production of cellulase cocktail was achieved via orthogonal experiment in a system with pH 6.0, 30 °C, and 96 h. Cellulase cocktail from P. oxalicum G2 can directly saccharify the extraction RIR, thereby achieving a maximum reducing sugar yield of 7.2 mg/mL, which is 1.7-fold higher than those of commercial cellulases. Results illustrate the potential of P. oxalicum G2 for enzymatic in situ saccharification.

Asunto(s)

RESUMEN

Sugarcane bagasse (SCB) is an abundant, renewable and inexpensive agricultural byproduct for the production of biofuel and other biobased products. To effectively saccharify SCB with cellulases, combination with dilute alkali salts Na2SO3/Na3PO4 (0.4% Na3PO4, 0.03% Na2SO3) at 7.5% sulfidity and hot water (DASHW) in "one-pot" pretreatment media by autoclaving at 110°C for 40min was attempted to pretreat SCB in this study. Furthermore, FT-IR, XRD and SEM were employed to characterize the changes in the cellulose structural characteristics (porosity, morphology, and crystallinity) of the pretreated Na2SO3/Na3PO4-SCB solid residue, which indicated that combination pretreatment could effectively remove lignin and hemicellulose for enhancing enzymatic saccharification. After 72h, the reducing sugars and glucose from the enzymatic in situ hydrolysis of 50g/L Na2SO3/Na3PO4-SCB in dilute Na2SO3/Na3PO4 (0.27% Na3PO4, 0.02% Na2SO3) media were obtained at 33.8 and 21.8g/L, respectively. Finally, the SCB-hydrolysates containing 20g/L glucose were used for ethanol fermentation in the presence of dilute alkali salts. After 48h, the ethanol yield was 0.42g ethanol/g glucose, which represents 82.1% of the theoretical yield. In conclusion, this study provided an effective pretreatment strategy for enhancing SCB's saccharification, which has potential application of other lignocellulosic materials.

Asunto(s)

RESUMEN

In this study, dilute alkali salts (0.6% NaClO, 0.067% Na2S) pretreatment at 10% sulfidity under the autoclave system at 120°C for 40min was used for pretreating bamboo shoot shell (BSS). Furthermore, FT-IR, XRD and SEM were employed to characterize the changes in the cellulose structural characteristics (porosity, morphology, and crystallinity) of the pretreated BSS solid residue. After 72h, the reducing sugars and glucose from the enzymatic in situ hydrolysis of 50g/L pretreated BSS in dilute NaClO/Na2S media could be obtained at 31.11 and 20.32g/L, respectively. Finally, the obtained BSS-hydrolysates containing alkalic salt NaClO/Na2S resulted in slightly negative effects on the ethanol production. Glucose in BSS-hydrolysates was fermented from 20.0 to 0.17g/L within 48h, and an ethanol yield of 0.41g/g glucose, which represents 80.1% of the theoretical yield, was obtained. This study provided an effective strategy for potential utilization of BSS.

Asunto(s)

RESUMEN

Aspergillus niger is an important microorganism that has been used for decades to produce extracellular enzymes. In this study, a novel Aspergillus niger strain integrated with a eukaryotic expression vector harboring the gpd-Shi promoter of shiitake mushrooms and cellulase gene of Ampullaria gigas Spix was engineered to improve cellulase production for the achievement of highly efficient saccharification of agricultural residues. In one strain, designated ACShi27, which exhibited the highest total cellulase expression, total cellulase, endoglucanase, exoglucanase, and xylanase expression levels were 1.73, 16.23, 17.73, and 150.83 U ml-1, respectively; these values were 14.5, 22.3, 24.6, and 17.3% higher than those of the wild-type Aspergillus niger M85 using wheat bran as an induction substrate. Production of cellulases and xylanase by solid-state fermentation followed by in situ saccharification of ACShi27 was investigated with alkaline-pretreated rice straw as a substrate. After 2 days of enzyme induction at 30 °C, followed by 48 h of saccharification at 50 °C, the conversion rate of carbon polymers into reducing sugar reached 293.2 mg g-1, which was 1.23-fold higher than that of the wild-type strain. The expression of sestc in Aspergillus niger can improve the total cellulase and xylanase activity and synergism, thereby enhancing the lignocellulose in situ saccharification.

RESUMEN

In this study, it was the first time to report that the cellulases of Galactomyces sp. CCZU11-1 showed high activity and stability in the culture and reaction media containing IL [Mmim]DMP. Using untreated chestnut shell (CNS) as carbon source in the culture media containing IL [Mmim]DMP (5%, w/v), high activity of FPA (28.6U/mL), xylanase (186.2U/mL), and CMCase (107.3U/mL) were obtained, and 184.9mg/L of total protein was achieved. Furthermore, the changes in the structural features (crystallinity, morphology, and porosity) of the solid residue of CNS utilized with Galactomyces sp. CCZU11-1 were characterized with Fourier transform infrared spectroscopy, scanning electron microscopy, and X-ray diffraction. After was enzymatically hydrolyzed with the prepared crude enzymes in IL diluted to 20% (w/v), a high yield of reducing sugars, 62.1%, was obtained. Significantly, Galactomyces sp. CCZU11-1 showed high potential for the efficient transformation of lignocellulosic materials to glucose in a single-step process.

Asunto(s)

RESUMEN

Choline acetate (ChOAc), a cholinium ionic liquid (IL), showed almost the same bagasse pretreatment capability as 1-ethyl-3-methylimidazolium acetate (EmimOAc), a conventional imidazolium IL used for biomass pretreatment. Moreover, ChOAc showed less of an inhibitory effect on cellulase than EmimOAc. Thus, ChOAc was used for IL/ultrasound-assisted pretreatment and in situ enzymatic saccharification, where IL was not washed out from the pretreated bagasse but diluted with the addition of a buffer solution. When in situ saccharification was performed for 48h in the presence of 10% ChOAc, the cellulose and hemicellulose saccharification percentages were 80% and 72%, respectively. When ChOAc was increased to 20%, the saccharification percentages were 72% and 53%, respectively. However, the values were just 28% and 2%, respectively, in case of 20% EmimOAc. A glucose/xylose solution free from IL and ChOAc aqueous solution without these sugars could be recovered separately by electrodialysis of the hydrolysate of in situ saccharification.

Asunto(s)