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"Saccharification"
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Conversion of Starchy Waste Streams into Polyhydroxyalkanoates Using Cupriavidus necator DSM 545
by
Basaglia, Marina
,
Favaro, Lorenzo
,
Parro, Elettra
in
Biodegradability
,
Bioplastics
,
By products
2020
Due to oil shortage and environmental problems, synthetic plastics have to be replaced by different biodegradable materials. A promising alternative could be polyhydroxyalkanoates (PHAs), and the low-cost abundant agricultural starchy by-products could be usefully converted into PHAs by properly selected and/or developed microbes. Among the widely available starchy waste streams, a variety of residues have been explored as substrates, such as broken, discolored, unripe rice and white or purple sweet potato waste. Cupriavidus necator DSM 545, a well-known producer of PHAs, was adopted in a simultaneous saccharification and fermentation (SSF) process through an optimized dosage of the commercial amylases cocktail STARGEN™ 002. Broken rice was found to be the most promising carbon source with PHAs levels of up to 5.18 g/L. This research demonstrates that rice and sweet potato waste are low-cost feedstocks for PHAs production, paving the way for the processing of other starchy materials into bioplastics.
Journal Article
Relative Saccharification of Sawdust Materials at Different Incubation pH-values
by
Seeletse, J. B. M.
,
van Wyk, J. P. H.
,
Ndukwe, N. A.
in
sawdust, cellulose, cellulase, delignification, saccharification
2025
The uncontrolled production of waste is a daily phenomenon that is experienced by most global communities, and the situation worsens due to the lack of effective waste management procedures. Solid waste such as sawdust is primarily produced by the forestry industry and although it is utilized by certain countries as briquettes to make fire or as an absorbent to clean fluid spillage as well as a component of ceilings, most of the sawdust along the Lagos Lagoon in Nigeria is left unattended as waste, contributing to environmental pollution. Cellulose, composed of glucose units is a structural component of sawdust and when saccharified the resulting glucose can be fermented into renewable substances such as bio-ethanol. The cellulose degradation process can be performed with a cellulase enzyme such as available in the fungus Aspergillus niger and during the current investigation, this enzyme system was used to bio-convert the cellulose component of sawdust from ten different trees along the Lagoon into glucose. To increase the cellulase action all sawdust materials were delignified before cellulase action with the main aim of determining the optimum pH value for maximum degradation of the various sawdust materials. The pH-related saccharification profile of each type of sawdust was constructed as well as the relative percentage of saccharification and it was concluded that all the materials were optimum degraded at acidic pH-values which varied between pH 5.0 and pH 6.0 that are like optimum pH-values reported for the other types of cellulose materials.
Journal Article
Effects of feruloyl-CoA 6'-hydroxylase 1 overexpression on lignin and cell wall characteristics in transgenic hybrid aspen
2025
In plant cell walls, lignin, cellulose, and the hemicelluloses form intricate three-dimensional structures. Owing to its complexity, lignin often acts as a bottleneck for the efficient utilization of polysaccharide components as biochemicals and functional materials. A promising approach to mitigate and/or overcome lignin recalcitrance is the qualitative and quantitative modification of lignin by genetic engineering. Feruloyl-CoA 6'-hydroxylase (F6'H1) is a 2-oxoglutarate-dependent dioxygenase that catalyzes the conversion of feruloyl-CoA, one of the intermediates of the lignin biosynthetic pathway, into 6'-hydroxyferuloyl-CoA, the precursor of scopoletin (7-hydroxy-6-methoxycoumarin). In a previous study with Arabidopsis thaliana, we demonstrated that overexpression of F6'H1 under a xylem-preferential promoter led to scopoletin incorporation into the cell wall. This altered the chemical structure of lignin without affecting lignin content or saccharification efficiency. In the present study, the same F6'H1 construct was introduced into hybrid aspen (Populus tremula × tremuloides T89), a model woody plant, and its effects on plant morphology, lignin chemical structure, global gene expression, and phenolic metabolism were examined. The transgenic plants successfully overproduced scopoletin while exhibiting severe growth retardation, a phenotype not previously observed in Arabidopsis. Scopoletin accumulation was most pronounced in the secondary walls of tracheary elements and the compound middle lamella, with low levels in the fiber cell walls. Overexpression of F6'H1 also affected the metabolism of aromatics, including lignin precursors. Heteronuclear single-quantum coherence (HSQC) NMR spectroscopy revealed that scopoletin in cell walls was bound to lignin, leading to a reduction in lignin content and changes in its monomeric composition and molar mass distribution. Furthermore, the enzymatic saccharification efficiency of the transgenic cell walls was more than three times higher than that of the wild-type plants, even without pretreatment. Although addressing growth inhibition remains a priority, incorporating scopoletin into lignin demonstrates significant potential for improving woody biomass utilization.
Journal Article
Understanding the structural and chemical changes of plant biomass following steam explosion pretreatment
2017
Background Biorefining of lignocellulosic biomass has become one of the most valuable alternatives for the production of multi-products such as biofuels. Pretreatment is a prerequisite to increase the enzymatic conversion of the recalcitrant lignocellulose. However, there is still considerable debate regarding the key features of biomass impacting the cellulase accessibility. In this study, we evaluate the structural and chemical features of three different representative biomasses (Miscanthus × giganteus, poplar and wheat straw), before and after steam explosion pretreatment at increasing severities, by monitoring chemical analysis, SEM, FTIR and 2D NMR. Results Regardless the biomass type, combined steam explosion pretreatment with dilute sulfuric acid impregnation resulted in significant improvement of the cellulose conversion. Chemical analyses revealed that the pretreatment selectively degraded the hemicellulosic fraction and associated cross-linking ferulic acids. As a result, the pretreated residues contained mostly cellulosic glucose and lignin. In addition, the pretreatment directly affected the cellulose crystallinity but these variations were dependent upon the biomass type. Important chemical modifications also occurred in lignin since the β-O-4' aryl-ether linkages were found to be homolytically cleaved, followed by some recoupling/recondensation to β-β' and β-5' linkages, regardless the biomass type. Finally, 2D NMR analysis of the whole biomass showed that the pretreatment preferentially degraded the syringyl-type lignin fractions in miscanthus and wheat straw while it was not affected in the pretreated poplar samples. Conclusions Our findings provide an enhanced understanding of parameters impacting biomass recalcitrance, which can be easily generalized to both woody and non-woody biomass species. Results indeed suggest that the hemicellulose removal accompanied by the significant reduction in the cross-linking phenolic acids and the redistribution of lignin are strongly correlated with the enzymatic saccharification, by loosening the cell wall structure thus allowing easier cellulase accessibility. By contrast, we have shown that the changes in the syringyl/guaiacyl ratio and the cellulose crystallinity do not seem to be relevant factors in assessing the enzymatic digestibility. Some biomass type-dependent and easily measurable FTIR factors are highly correlated to saccharification.
Journal Article
A finalized determinant for complete lignocellulose enzymatic saccharification potential to maximize bioethanol production in bioenergy Miscanthus
2019
Background Miscanthus is a leading bioenergy crop with enormous lignocellulose production potential for biofuels and chemicals. However, lignocellulose recalcitrance leads to biomass process difficulty for an efficient bioethanol production. Hence, it becomes essential to identify the integrative impact of lignocellulose recalcitrant factors on cellulose accessibility for biomass enzymatic hydrolysis. In this study, we analyzed four typical pairs of Miscanthus accessions that showed distinct cell wall compositions and sorted out three major factors that affected biomass saccharification for maximum bioethanol production. Results Among the three optimal (i.e., liquid hot water, H2SO4 and NaOH) pretreatments performed, mild alkali pretreatment (4% NaOH at 50 °C) led to almost complete biomass saccharification when 1% Tween-80 was co-supplied into enzymatic hydrolysis in the desirable Miscanthus accessions. Consequently, the highest bioethanol yields were obtained at 19% (% dry matter) from yeast fermentation, with much higher sugar–ethanol conversion rates by 94–98%, compared to the other Miscanthus species subjected to stronger pretreatments as reported in previous studies. By comparison, three optimized pretreatments distinctively extracted wall polymers and specifically altered polymer features and inter-linkage styles, but the alkali pretreatment caused much increased biomass porosity than that of the other pretreatments. Based on integrative analyses, excellent equations were generated to precisely estimate hexoses and ethanol yields under various pretreatments and a hypothetical model was proposed to outline an integrative impact on biomass saccharification and bioethanol production subjective to a predominate factor (CR stain) of biomass porosity and four additional minor factors (DY stain, cellulose DP, hemicellulose X/A, lignin G-monomer). Conclusion Using four pairs of Miscanthus samples with distinct cell wall composition and varied biomass saccharification, this study has determined three main factors of lignocellulose recalcitrance that could be significantly reduced for much-increased biomass porosity upon optimal pretreatments. It has also established a novel standard that should be applicable to judge any types of biomass process technology for high biofuel production in distinct lignocellulose substrates. Hence, this study provides a potential strategy for precise genetic modification of lignocellulose in all bioenergy crops.
Journal Article
Combination of ultrasonication and deep eutectic solvent in pretreatment of lignocellulosic biomass for enhanced enzymatic saccharification
2021
The synergistic effect of ultrasonication with deep eutectic solvent (DES) on pretreatment of oil palm empty fruit bunch (OPEFB) was investigated. Three different types of DESs, namely choline chloride:lactic acid (ChCl:LA), choline chloride:urea (ChCl:U) and choline chloride:glycerol (ChCl:G) were applied. The performance of the pretreatment was evaluated based on yield of reducing sugars, lignin content, crystallinity index, structural and morphology changes. ChCl:LA pretreated OPEFB attained the highest yield of reducing sugars (36.7%) under the action of ultrasonication for 15 min at sonication power 60% (210 W) and temperature 50 °C, followed by ChCl:U (35.8%) and ChCl:G (35.3%). Under these conditions, ChCl:LA pretreated OPEFB showed significant change in structural and morphology, associated with the lowest crystallinity and lignin content. ChCl:LA promoted the pretreatment process in view of its intrinsic properties of low viscosity and low surface tension. The incorporation of ultrasonication in DES pretreatment significantly increased the reducing sugars yield suggested the synergistic effect of ultrasonication with DES pretreatment. These signifies that ultrasound-assisted DES pretreatment could be a promising alternative pretreatment technique for lignocellulosic biomass.Graphic abstract
Journal Article
Recent advances in bioethanol production from lignocelluloses: a comprehensive review with a focus on enzyme engineering and designer biocatalysts
by
Chandel, Anuj K
,
Kumar, Sachin
,
Prabhu, Ashish A
in
auxiliary proteins
,
Biocatalysts
,
Biofuels
2020
Many countries have their biofuel policy programs in place as part of their overall strategy to achieve sustainable development. Among biofuels, bioethanol as a promising alternative to gasoline is of substantial interest. However, there is limited availability of a sufficient quantity of bioethanol to meet demands due to bottlenecks in the present technologies to convert non-edible feedstocks, including lignocelluloses. This review article presents and critically discusses the recent advances in the pretreatment of lignocellulosic biomass, with a focus on the use of green solvents, including ionic liquids and deep eutectic solvents, followed by enzymatic saccharification using auxiliary proteins for the efficient saccharification of pretreated biomass. Different techniques used in strain improvement strategies to develop hyper-producing deregulated lignocellulolytic strains are also compared and discussed. The advanced techniques employed for fermentation of mixed sugars contained in lignocellulosic hydrolysates for maximizing bioethanol production are summarized with an emphasis on pathway and transporters engineering for xylose assimilation. Further, the integration of different steps is suggested and discussed for efficient biomass utilization and improved ethanol yields and productivity.
Journal Article
OsCESA9 conserved‐site mutation leads to largely enhanced plant lodging resistance and biomass enzymatic saccharification by reducing cellulose DP and crystallinity in rice
2017
Summary Genetic modification of plant cell walls has been posed to reduce lignocellulose recalcitrance for enhancing biomass saccharification. Since cellulose synthase (CESA) gene was first identified, several dozen CESA mutants have been reported, but almost all mutants exhibit the defective phenotypes in plant growth and development. In this study, the rice (Oryza sativa) Osfc16 mutant with substitutions (W481C, P482S) at P‐CR conserved site in CESA9 shows a slightly affected plant growth and higher biomass yield by 25%–41% compared with wild type (Nipponbare, a japonica variety). Chemical and ultrastructural analyses indicate that Osfc16 has a significantly reduced cellulose crystallinity (CrI) and thinner secondary cell walls compared with wild type. CESA co‐IP detection, together with implementations of a proteasome inhibitor (MG132) and two distinct cellulose inhibitors (Calcofluor, CGA), shows that CESA9 mutation could affect integrity of CESA4/7/9 complexes, which may lead to rapid CESA proteasome degradation for low‐DP cellulose biosynthesis. These may reduce cellulose CrI, which improves plant lodging resistance, a major and integrated agronomic trait on plant growth and grain production, and enhances biomass enzymatic saccharification by up to 2.3‐fold and ethanol productivity by 34%–42%. This study has for the first time reported a direct modification for the low‐DP cellulose production that has broad applications in biomass industries.
Journal Article
Lytic xylan oxidases from wood-decay fungi unlock biomass degradation
2018
Wood biomass is the most abundant feedstock envisioned for the development of modern biorefineries. However, the cost-effective conversion of this form of biomass into commodity products is limited by its resistance to enzymatic degradation. Here we describe a new family of fungal lytic polysaccharide monooxygenases (LPMOs) prevalent among white-rot and brown-rot basidiomycetes that is active on xylans--a recalcitrant polysaccharide abundant in wood biomass. Two AA14 LPMO members from the white-rot fungus Pycnoporus coccineus substantially increase the efficiency of wood saccharification through oxidative cleavage of highly refractory xylan-coated cellulose fibers. The discovery of this unique enzyme activity advances our knowledge on the degradation of woody biomass in nature and offers an innovative solution for improving enzyme cocktails for biorefinery applications.
Journal Article
Fermentation and microbial characteristics of sorghum-integrated Makgeolli using different saccharification agents Koji and Nuruk
by
Chen Chen
,
Marianne T. Howell
,
Tadesse Fikre Teferra
in
fermentation
,
Makgeolli
,
microbial dynamics
2026
This study evaluated sorghum as a partial substitute for rice in Makgeolli brewing and compared fermentation characteristics between two saccharification agents, koji and nuruk. Makgeolli was prepared with 75% rice and 25% sorghum, and physicochemical properties and microbial communities were analyzed using culture-dependent and culture-independent (16S rRNA and ITS sequencing) methods. During fermentation, pH decreased below 3.5 in all treatments. Lactic acid bacteria exceeded 7 log CFU/mL in nuruk Makgeolli but were not detected in koji Makgeolli by culture-based methods, whereas 16S rRNA sequencing identified Firmicutes and lactic acid bacteria in both fermentations. ITS analysis showed Aspergillus spp. dominance in koji and Saccharomycopsis fibuligera in nuruk. Sorghum substitution did not significantly affect acidification or microbial succession. These results demonstrate that sorghum can be incorporated into Makgeolli without altering fermentation behavior and highlight the value of combining culture-dependent and sequencing approaches to characterize microbial dynamics.
Journal Article