saccharification
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Bioethanol production from lignocellulosic materials is hindered by the high costs of pretreatment and the enzymes. The present study aimed to evaluate whether co-cultivation of four selected cellulolytic fungi yields higher cellulase and xylanase activities compared to the monocultures and to investigate whether the enzymes from the co-cultures yield higher saccharification on selected plant materials without thermo-chemical pretreatment. The fungal isolates, Trichoderma reesei F118, Penicillium javanicum FS7, Talaromyces sp. F113, and Talaromyces pinophilus FM9, were grown as monocultures and binary co-cultures under submerged conditions for 7 days. The cellulase and xylanase activities of the culture filtrates were measured, and the culture filtrates were employed for the saccharification of sugarcane leaves, Guinea grass leaves, and water hyacinth stems and leaves. Total reducing sugars and individual sugars released from each plant material were quantified. The co-culture of Talaromyces sp. F113 with Penicillium javanicum FS7 and of T. reesei F118 with T. pinophilus FM9 produced significantly higher cellulase activities compared to the corresponding monocultures whereas no effect was observed on xylanase activities. Overall, the highest amounts of total reducing sugars and individual sugars were obtained from Guinea grass leaves saccharified with the co-culture of T. reesei F118 with T. pinophilus FM9, yielding 63.5% saccharification. Guinea grass leaves were found to be the most susceptible to enzymatic saccharification without pre-treatment, while water hyacinth stems and leaves were the least. Accordingly, the study suggests that fungal co-cultivation could be a promising approach for the saccharification of lignocellulosic materials for bioethanol production.
- MeSH
- celulasa * metabolismus MeSH
- endo-1,4-beta-xylanasy metabolismus MeSH
- ethanol metabolismus MeSH
- Hypocreales enzymologie metabolismus růst a vývoj MeSH
- kokultivační techniky * MeSH
- lignin * metabolismus MeSH
- listy rostlin mikrobiologie MeSH
- Penicillium * enzymologie metabolismus růst a vývoj MeSH
- Saccharum * mikrobiologie metabolismus MeSH
- Talaromyces * enzymologie metabolismus růst a vývoj MeSH
- Publikační typ
- časopisecké články MeSH
Endo-glucanase (cellulase) and xylanase have high industrial demand due to their vast application in industrial processes. This study reports statistical based experimental optimization for co-production of endo-glucanase and xylanase from Bacillus sonorensis BD92. Response surface methodology (RSM) involving central composite design (CCD) with full factorial experiments (23) was applied to elucidate the components that significantly affect co-production of endo-glucanase and xylanase. The optimum co-production conditions for endo-glucanase and xylanase were as follows: carboxymethyl cellulose (CMC) 20 g/L, yeast extract 15 g/L, and time 72 h. The maximum endo-glucanase and xylanase production obtained was 1.46 and 5.69 U/mL, respectively, while the minimum endo-glucanase and xylanase production obtained was 0.66 and 0.25 U/mL, respectively. This statistical model was efficient because only 20 experimental runs were necessary to assess the highest production conditions, and the model accuracy was very satisfactory as coefficient of determination (R2) was 0.95 and 0.89 for endo-glucanase and xylanase, respectively. Further, potential application of these enzymes for saccharification of lignocellulosic biomass (wheat bran, wheat straw, rice straw, and cotton stalk) was also investigated. The results revealed that the biomass was susceptible to enzymatic saccharification and the amount of reducing sugars (glucose and xylose) increased with increase in incubation time. In conclusion, Bacillus sonorensis BD92 reveals a promise as a source of potential endo-glucanase and xylanase producer that could be useful for degrading plant biomass into value-added products of economic importance using precise statistically optimized conditions.
- MeSH
- Bacillus růst a vývoj metabolismus MeSH
- biomasa * MeSH
- celulasa biosyntéza MeSH
- endo-1,4-beta-xylanasy biosyntéza MeSH
- fermentace MeSH
- hydrolýza MeSH
- průmyslová mikrobiologie metody MeSH
- rýže (rod) metabolismus MeSH
- sodná sůl karboxymethylcelulosy MeSH
- statistické modely MeSH
- Publikační typ
- časopisecké články MeSH
This review provides current information on the production of ethanol from lignocellulosic biomass, with the main focus on relationships between process design and efficiency, expressed as ethanol concentration, yield and productivity. In spite of unquestionable advantages of lignocellulosic biomass as a feedstock for ethanol production (availability, price, non-competitiveness with food, waste material), many technological bottlenecks hinder its wide industrial application and competitiveness with 1st generation ethanol production. Among the main technological challenges are the recalcitrant structure of the material, and thus the need for extensive pretreatment (usually physico-chemical followed by enzymatic hydrolysis) to yield fermentable sugars, and a relatively low concentration of monosaccharides in the medium that hinder the achievement of ethanol concentrations comparable with those obtained using 1st generation feedstocks (e.g. corn or molasses). The presence of both pentose and hexose sugars in the fermentation broth, the price of cellulolytic enzymes, and the presence of toxic compounds that can inhibit cellulolytic enzymes and microbial producers of ethanol are major issues. In this review, different process configurations of the main technological steps (enzymatic hydrolysis, fermentation of hexose/and or pentose sugars) are discussed and their efficiencies are compared. The main features, benefits and drawbacks of simultaneous saccharification and fermentation (SSF), simultaneous saccharification and fermentation with delayed inoculation (dSSF), consolidated bioprocesses (CBP) combining production of cellulolytic enzymes, hydrolysis of biomass and fermentation into one step, together with an approach combining utilization of both pentose and hexose sugars are discussed and compared with separate hydrolysis and fermentation (SHF) processes. The impact of individual technological steps on final process efficiency is emphasized and the potential for use of immobilized biocatalysts is considered.
Enzymatic hydrolysis of waste paper is becoming a perspective way to obtain raw material for production of liquid biofuels. Reducing sugars solutions that arise from the process of saccharification are a precursors for following or simultaneous fermentation to ethanol. Different types of waste paper were evaluated, in terms of composition and usability, in order to select the appropriate type of the waste paper for the enzymatic hydrolysis process. Novozymes® enzymes NS50013 and NS50010 were used in a laboratory scale trials. Technological conditions, which seem to be the most suitable for hydrolysis after testing on cellulose pulp and filter paper, were applied to hydrolysis of widely available waste papers - offset paper, cardboard, recycled paper in two qualities, matte MYsol offset paper and for comparison again on model materials. The highest yields were achieved for the cardboard, which was further tested using various pretreatment combinations in purpose of increasing the hydrolysis yields.
In this work hydrothermally pretreated wheat straw was used for production of bioethanol by Saccharomyces cerevisiae and carotene-enriched biomass by red yeasts Rhodotorula glutinis, Cystofilobasidium capitatum and Sporobolomyces roseus. To evaluate the convertibility of pretreated wheat straw into ethanol, simultaneous saccharification and fermentation of S. cerevisiae was performed under semi-anaerobic conditions. The highest ethanol production efficiency of 65-66% was obtained following pretreatment at 200°C without the catalytic action of acetic acid, and at 195 and 200°C respectively in the presence of catalyst. Red yeast strain S. roseus produced 1.73-2.22 mg g(-1) of ergosterol on the filter cake, 1.15-4.17 mg g(-1) of ergosterol and 1.23-1.56 mg g(-1) of β-carotene on pretreated wheat straw hydrolysates and also the highest amount of carotenoids and ergosterol on untreated wheat straw (1.70 and 4.17 mg g(-1), respectively).
- MeSH
- biomasa * MeSH
- biopaliva mikrobiologie MeSH
- bioreaktory mikrobiologie MeSH
- biotechnologie metody MeSH
- ergosterol biosyntéza MeSH
- ethanol metabolismus MeSH
- glukosa metabolismus MeSH
- hydrolýza účinky léků MeSH
- karotenoidy metabolismus MeSH
- kvasinky účinky léků růst a vývoj metabolismus MeSH
- odpadní produkty analýza MeSH
- pšenice účinky léků MeSH
- sacharidy chemie MeSH
- teplota * MeSH
- voda farmakologie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Waste paper belongs to a group of quantitatively the most produced waste types. Enzymatic hydrolysis is becoming a suitable way to treat this type of waste and at the same time, to produce a valuable liquid biofuel, because reducing sugars solutions that are formed during the process of saccharification can be a precursor for following or simultaneous fermentation. If it will be possible to make the enzymatic hydrolysis of the waste paper economically viable, it could serve as one of the new ways to lower the dependence of the transport sector on oil in the future. Only several studies comparing the enzymatic hydrolysis of different waste papers were performed in the past; they are summarized in this manuscript. In our experimental trials, suitable technological conditions for waste paper enzymatic hydrolysis using enzymes from Novozymes® biomass kit: enzymes NS50013 and NS50010 were investigated. The following enzymatic hydrolysis parameters in laboratory scale trials were verified on high cellulose content substrates-filter paper and cellulose pulp: type of buffer, pH, temperature, concentration of the substrate, loading of the enzyme and rate of stirring.
Lytic polysaccharide monooxygenases (LPMOs) are industrially important oxidoreductases employed in lignocellulose saccharification. Using advanced time-resolved mass spectrometric techniques, we elucidated the structural determinants for substrate-mediated stabilization of the fungal LPMO9C from Neurosporacrassa during catalysis. LPMOs require a reduction in the active-site copper for catalytic activity. We show that copper reduction in NcLPMO9C leads to structural rearrangements and compaction around the active site. However, longer exposure to the reducing agent ascorbic acid also initiated an uncoupling reaction of the bound oxygen species, leading to oxidative damage, partial unfolding, and even fragmentation of NcLPMO9C. Interestingly, no changes in the hydrogen/deuterium exchange rate were detected upon incubation of oxidized or reduced LPMO with crystalline cellulose, indicating that the LPMO-substrate interactions are mainly side-chain mediated and neither affect intraprotein hydrogen bonding nor induce significant shielding of the protein surface. On the other hand, we observed a protective effect of the substrate, which slowed down the autooxidative damage induced by the uncoupling reaction. These observations further complement the picture of structural changes during LPMO catalysis.
- MeSH
- celulosa chemie MeSH
- fungální proteiny chemie MeSH
- hmotnostní spektrometrie s elektrosprejovou ionizací MeSH
- hmotnostní spektrometrie MeSH
- katalytická doména MeSH
- katalýza MeSH
- koncentrace vodíkových iontů MeSH
- konformace proteinů MeSH
- kyslík chemie MeSH
- lignin chemie MeSH
- měď chemie MeSH
- Neurospora crassa enzymologie MeSH
- oxidační stres MeSH
- oxidoreduktasy chemie MeSH
- oxygenasy se smíšenou funkcí chemie MeSH
- polysacharidy chemie MeSH
- reaktivní formy kyslíku chemie MeSH
- substrátová specifita MeSH
- vazba proteinů MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
BACKGROUND: QTL cloning for the discovery of genes underlying polygenic traits has historically been cumbersome in long-lived perennial plants like Populus. Linkage disequilibrium-based association mapping has been proposed as a cloning tool, and recent advances in high-throughput genotyping and whole-genome resequencing enable marker saturation to levels sufficient for association mapping with no a priori candidate gene selection. Here, multiyear and multienvironment evaluation of cell wall phenotypes was conducted in an interspecific P. trichocarpa x P. deltoides pseudo-backcross mapping pedigree and two partially overlapping populations of unrelated P. trichocarpa genotypes using pyrolysis molecular beam mass spectrometry, saccharification, and/ or traditional wet chemistry. QTL mapping was conducted using a high-density genetic map with 3,568 SNP markers. As a fine-mapping approach, chromosome-wide association mapping targeting a QTL hot-spot on linkage group XIV was performed in the two P. trichocarpa populations. Both populations were genotyped using the 34 K Populus Infinium SNP array and whole-genome resequencing of one of the populations facilitated marker-saturation of candidate intervals for gene identification. RESULTS: Five QTLs ranging in size from 0.6 to 1.8 Mb were mapped on linkage group XIV for lignin content, syringyl to guaiacyl (S/G) ratio, 5- and 6-carbon sugars using the mapping pedigree. Six candidate loci exhibiting significant associations with phenotypes were identified within QTL intervals. These associations were reproducible across multiple environments, two independent genotyping platforms, and different plant growth stages. cDNA sequencing for allelic variants of three of the six loci identified polymorphisms leading to variable length poly glutamine (PolyQ) stretch in a transcription factor annotated as an ANGUSTIFOLIA C-terminus Binding Protein (CtBP) and premature stop codons in a KANADI transcription factor as well as a protein kinase. Results from protoplast transient expression assays suggested that each of the polymorphisms conferred allelic differences in the activation of cellulose, hemicelluloses, and lignin pathway marker genes. CONCLUSION: This study illustrates the utility of complementary QTL and association mapping as tools for gene discovery with no a priori candidate gene selection. This proof of concept in a perennial organism opens up opportunities for discovery of novel genetic determinants of economically important but complex traits in plants.
- MeSH
- alely MeSH
- buněčná stěna genetika MeSH
- celulosa metabolismus MeSH
- fenotyp MeSH
- genetická vazba MeSH
- genotyp MeSH
- jednonukleotidový polymorfismus MeSH
- lignin biosyntéza MeSH
- lod skóre MeSH
- lokus kvantitativního znaku MeSH
- mapování chromozomů MeSH
- Populus genetika MeSH
- rostlinné geny * MeSH
- rostlinné proteiny chemie genetika MeSH
- sekvence nukleotidů MeSH
- sekvenční seřazení MeSH
- transkripční faktory chemie genetika MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
