174 research outputs found
Machine learning reveals sequence-function relationships in family 7 glycoside hydrolases
Family 7 glycoside hydrolases (GH7) are among the principal enzymes for cellulose degradation in nature and industrially. These enzymes are often bimodular, including a catalytic domain and carbohydrate-binding module (CBM) attached via a flexible linker, and exhibit an active site that binds cello-oligomers of up to ten glucosyl moieties. GH7 cellulases consist of two major subtypes: cellobiohydrolases (CBH) and endoglucanases (EG). Despite the critical importance of GH7 enzymes, there remain gaps in our understanding of how GH7 sequence and structure relate to function. Here, we employed machine learning to gain data-driven insights into relation-ships between sequence, structure, and function across the GH7 family. Machine-learning models, trained only on the number of residues in the active-site loops as features, were able to discriminate GH7 CBHs and EGs with up to 99% ac-curacy, demonstrating that the lengths of loops A4, B2, B3, and B4 strongly correlate with functional subtype across the GH7 family. Classification rules were derived such that specific residues at 42 different sequence positions each predicted the functional subtype with accuracies surpassing 87%. A random forest model trained on residues at 19 positions in the catalytic domain predicted the presence of a CBM with 89.5% accuracy. Our machine learning results recapitulate, as top-performing features, a substantial number of the sequence positions determined by previous experimental studies to play vital roles in GH7 activity. We surmise that the yet-to-be-explored sequence positions among the top-performing features also contribute to GH7 functional variation and may be exploited to understand and manipulate function
Reductive Catalytic Fractionation of Corn Stover Lignin
Reductive catalytic fractionation (RCF) has emerged as an effective biomass pretreatment strategy to depolymerize lignin into tractable fragments in high yields. We investigate the RCF of corn stover, a highly abundant herbaceous feedstock, using carbon-supported Ru and Ni catalysts at 200 and 250 °C in methanol and, in the presence or absence of an acid cocatalyst (H₃PO₄ or an acidified carbon support). Three key performance variables were studied: (1) the effectiveness of lignin extraction as measured by the yield of lignin oil, (2) the yield of monomers in the lignin oil, and (3) the carbohydrate retention in the residual solids after RCF. The monomers included methyl coumarate/ferulate, propyl guaiacol/syringol, and ethyl guaiacol/syringol. The Ru and Ni catalysts performed similarly in terms of product distribution and monomer yields. The monomer yields increased monotonically as a function of time for both temperatures. At 6 h, monomer yields of 27.2 and 28.3% were obtained at 250 and 200 °C, respectively, with Ni/C. The addition of an acid cocatalysts to the Ni/C system increased monomer yields to 32% for acidified carbon and 38% for phosphoric acid at 200 °C. The monomer product distribution was dominated by methyl coumarate regardless of the use of the acid cocatalysts. The use of phosphoric acid at 200 °C or the high temperature condition without acid resulted in complete lignin extraction and partial sugar solubilization (up to 50%) thereby generating lignin oil yields that exceeded the theoretical limit. In contrast, using either Ni/C or Ni on acidified carbon at 200 °C resulted in moderate lignin oil yields of ca. 55%, with sugar retention values >90%. Notably, these sugars were amenable to enzymatic digestion, reaching conversions >90% at 96 h. Characterization studies on the lignin oils using two-dimensional heteronuclear single quantum coherence nuclear magnetic resonance and gel permeation chromatrography revealed that soluble oligomers are formed via solvolysis, followed by further fragmentation on the catalyst surface via hydrogenolysis. Overall, the results show that clear trade-offs exist between the levels of lignin extraction, monomer yields, and carbohydrate retention in the residual solids for different RCF conditions of corn stover.National Science Foundation (U.S.) (1454299
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Technoeconomic and life-cycle analysis of single-step catalytic conversion of wet ethanol into fungible fuel blendstocks
Technoeconomic and life-cycle analyses are presented for catalytic conversion of ethanol to fungible hydrocarbon fuel blendstocks, informed by advances in catalyst and process development. Whereas prior work toward this end focused on 3-step processes featuring dehydration, oligomerization, and hydrogenation, the consolidated alcohol dehydration and oligomerization (CADO) approach described here results in 1-step conversion of wet ethanol vapor (40 wt% in water) to hydrocarbons and water over a metal-modified zeolite catalyst. A development project increased liquid hydrocarbon yields from 36% of theoretical to >80%, reduced catalyst cost by an order of magnitude, scaled up the process by 300-fold, and reduced projected costs of ethanol conversion 12-fold. Current CADO products conform most closely to gasoline blendstocks, but can be blended with jet fuel at low levels today, and could potentially be blended at higher levels in the future. Operating plus annualized capital costs for conversion of wet ethanol to fungible blendstocks are estimated at 1.44/GJ in the future, similar to the unit energy cost of producing anhydrous ethanol from wet ethanol (100 per barrel but not at 60 per barrel. Life-cycle greenhouse gas emission reductions for CADO-derived hydrocarbon blendstocks closely follow those for the ethanol feedstock
Continuous succinic acid production by Actinobacillus succinogenes on xylose‑enriched hydrolysate
BACKGROUND : Bio-manufacturing of high-value chemicals in parallel to renewable biofuels has the potential to
dramatically improve the overall economic landscape of integrated lignocellulosic biorefineries. However, this will
require the generation of carbohydrate streams from lignocellulose in a form suitable for efficient microbial conversion
and downstream processing appropriate to the desired end use, making overall process development, along
with selection of appropriate target molecules, crucial to the integrated biorefinery. Succinic acid (SA), a high-value
target molecule, can be biologically produced from sugars and has the potential to serve as a platform chemical for
various chemical and polymer applications. However, the feasibility of microbial SA production at industrially relevant
productivities and yields from lignocellulosic biorefinery streams has not yet been reported.
RESULTS : Actinobacillus succinogenes 130Z was immobilised in a custom continuous fermentation setup to produce
SA on the xylose-enriched fraction of a non-detoxified, xylose-rich corn stover hydrolysate stream produced from
deacetylation and dilute acid pretreatment. Effective biofilm attachment, which serves as a natural cell retention
strategy to increase cell densities, productivities and resistance to toxicity, was accomplished by means of a novel
agitator fitting. A maximum SA titre, yield and productivity of 39.6 g L−1, 0.78 g g−1 and 1.77 g L−1 h−1 were achieved,
respectively. Steady states were obtained at dilution rates of 0.02, 0.03, 0.04, and 0.05 h−1 and the stirred biofilm reactor
was stable over prolonged periods of operation with a combined fermentation time of 1550 h. Furthermore, it was
found that a gradual increase in the dilution rate was required to facilitate adaptation of the culture to the hydrolysate,
suggesting a strong evolutionary response to the toxic compounds in the hydrolysate. Moreover, the two primary
suspected fermentation inhibitors, furfural and HMF, were metabolised during fermentation with the concentration of
each remaining at zero across all steady states.
CONCLUSIONS : The results demonstrate that immobilised A. succinogenes has the potential for effective conversion
of an industrially relevant, biomass-derived feed stream to succinic acid. Furthermore, due to the attractive yields,
productivities and titres achieved in this study, the process has the potential to serve as a means for value-added
chemical manufacturing in the integrated biorefinery.The National Research Foundation
(NRF) and the US Department of Energy Bioenergy Technologies Office.http://biotechnologyforbiofuels.biomedcentral.comam201
Lignin-first biorefining of Nordic poplar to produce cellulose fibers could displace cotton production on agricultural lands
Here, we show that lignin-first biorefining of poplar can enable the production of dissolving cellulose pulp that can produce regenerated cellulose, which could substitute cotton. These results in turn indicate that agricultural land dedicated to cotton could be reclaimed for food production by extending poplar plantations to produce textile fibers. Based on climate-adapted poplar clones capable of growth on marginal lands in the Nordic region, we estimate an environmentally sustainable annual biomass production of similar to 11 tonnes/ha. At scale, lignin-first biorefining of this poplar could annually generate 2.4 tonnes/ha of dissolving pulp for textiles and 1.1 m(3) biofuels. Life cycle assessment indicates that, relative to cotton production, this approach could substantially reduce water consumption and identifies certain areas for further improvement. Overall, this work highlights a new value chain to reduce the environmental footprint of textiles, chemicals, and biofuels while enabling land reclamation and water savings from cotton back to food production
A promiscuous cytochrome P450 aromatic O-demethylase for lignin bioconversion
FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOMicrobial aromatic catabolism offers a promising approach to convert lignin, a vast source of renewable carbon, into useful products. Aryl-O-demethylation is an essential biochemical reaction to ultimately catabolize coniferyl and sinapyl lignin-derived a9FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOFAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO2013/08293-72014/10448-12016/22956-7We acknowledge funding from NSF grants to J.L.D. (MCB-1715176), K.N.H. (CHE-1361104), and E.L.N. (DEB-1556541 and MCB-1615365) and BBSRC grants to J.E.M. (BB/P011918/1, BB/L001926/1 and a studentship to S.J.B.M.). G.T.B., M.M.M., C.W.J., M.F.C., E.L.N.,
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