Brachytic2 mutation is able to counteract the main pleiotropic effects of brown midrib3 mutant in maize.

Maize is the basis of nutrition of domesticated herbivores and one of the most promising energy crops. The presence of lignin in the cell wall, tightly associated to carbohydrates, prevents the physical access of enzymes such as cellulase, limiting the carbohydrate degradability and consequently the energy value. To increase the utilization of the biomass cellulose content, the challenge of breeding programs is to lower or modify the lignin components. In maize several mutations are able to modify the lignin content and in particular the mutation in brown midrib3 (bm3) gene appeared as one of the most promising in breeding programs. Unfortunately this mutation has several negative pleiotropic effects on various important agronomic traits such as stay green, lodging and susceptibility to several infections.The maize Brachyitic 2 (br2) gene encodes for a putative protein involved in polar movement of auxins. br2 mutant plants are characterized by shortening of lower stalk internodes, unusual stalk strength and tolerance to wind lodging, darker leaves persisting longer in the active green state in comparison to wild type plants, suggesting a possible utilization of br2 plants to counteract the negative effects of the bm3 mutation. In this work, we report the generation and a preliminary characterization of the double mutant bm3 br2, suggesting the potential use of this new genetic material to increase biomass cellulose utilization.

Diversifying the products from the organic fraction of municipal solid waste (OFMSW) by producing polyhydroxyalkanoates from the liquid fraction and biomethane from the residual solid fraction.

This study describes the diversification of products obtainable from the organic fraction of municipal solid waste (OFMSW) by producing polyhydroxyalkanoates (PHA) from the liquid fraction and biomethane from the residual solid fraction. OFMSW samples were taken during the 2021 season from two full field scale plants treating wastes. After solid/liquid (S/L) separation, 80% of initial organic acids (OAs) were released in the liquid stream. OAs were then used as feed for PHA production and residual solid cakes were tested for biomethane production. Complete mass balance and energy balance were calculated. PHAs production was of 115 ± 23 (n = 6) g kg- 1 OFMSW (TS) and residual biomethane of 219 ± 3 g kg- 1 OFMSW TS, (n = 6). Energy balance indicated that nearly 40% of OFMSW energy was recovered as products. This value was lower than that obtained previously when AD was performed before OAs separation (i.e. 64%).

Variation in sugarcane biomass composition and enzymatic saccharification of leaves, internodes and roots.

BACKGROUND: The composition of biomass determines its suitability for different applications within a biorefinery system. The proportion of the major biomass fractions (sugar, cellulose, hemicellulose and lignin) may vary in different sugarcane genotypes and growth environments and different parts of the plant. This study investigated the composition of mature and immature internodes, roots and mature leaves of sugarcane. RESULTS: Internodes were found to have a significantly larger alcohol-soluble component than leaves and roots. The primary difference between the immature and mature internodes was the ratio of soluble sugars. In mature tissues, sucrose content was significantly higher, whereas in immature internodal tissues there was lower sucrose and heightened concentrations of reducing sugars. Carbon (C) partitioning in leaf tissues was characterised by low levels of soluble components and high "other" and cell wall fractions. Root tissue had low ratios of soluble fractions relative to their cell wall contents, indicating a lack of storage of soluble carbon. There was no significant difference in the ratio of the major cell wall fractions between the major organ types. Characterisation of individual non-cellulosic monomers indicated leaf and root tissues had significantly higher arabinose and galactose fractions. Significantly larger proportions of syringyl lignin compounds and the hydroxycinnamic compound, p-coumaric acid were observed in mature internodal tissues compared to the other tissue types. Tissue-specific differences in composition were shown to greatly affect the recalcitrance of the cell wall to enzymatic saccharification. CONCLUSIONS: Overall, this study displayed clear evidence of the differential partitioning of C throughout the sugarcane plant in specific organs. These organ-specific differences have major implications in their utility as a bioproduct feedstock. For example, the inclusion of trash (leaves) with the culms (internodes) may alter processing efficiency.

Conversion of poplar biomass into high-energy density tricyclic sesquiterpene jet fuel blendstocks.

BACKGROUND: In an effort to ensure future energy security, reduce greenhouse gas emissions and create domestic jobs, the US has invested in technologies to develop sustainable biofuels and bioproducts from renewable carbon sources such as lignocellulosic biomass. Bio-derived jet fuel is of particular interest as aviation is less amenable to electrification compared to other modes of transportation and synthetic biology provides the ability to tailor fuel properties to enhance performance. Specific energy and energy density are important properties in determining the attractiveness of potential bio-derived jet fuels. For example, increased energy content can give the industry options such as longer range, higher load or reduced takeoff weight. Energy-dense sesquiterpenes have been identified as potential next-generation jet fuels that can be renewably produced from lignocellulosic biomass. RESULTS: We developed a biomass deconstruction and conversion process that enabled the production of two tricyclic sesquiterpenes, epi-isozizaene and prespatane, from the woody biomass poplar using the versatile basidiomycete Rhodosporidium toruloides. We demonstrated terpene production at both bench and bioreactor scales, with prespatane titers reaching 1173.6 mg/L when grown in poplar hydrolysate in a 2 L bioreactor. Additionally, we examined the theoretical fuel properties of prespatane and epi-isozizaene in their hydrogenated states as blending options for jet fuel, and compared them to aviation fuel, Jet A. CONCLUSION: Our findings indicate that prespatane and epi-isozizaene in their hydrogenated states would be attractive blending options in Jet A or other lower density renewable jet fuels as they would improve viscosity and increase their energy density. Saturated epi-isozizaene and saturated prespatane have energy densities that are 16.6 and 18.8% higher than Jet A, respectively. These results highlight the potential of R. toruloides as a production host for the sustainable and scalable production of bio-derived jet fuel blends, and this is the first report of prespatane as an alternative jet fuel.

Implementing polyhydroxyalkanoates production to anaerobic digestion of organic fraction of municipal solid waste to diversify products and increase total energy recovery.

A simple biorefinery aimed at producing both biomethane (CH4) and polyhydroxyalkanoates (PHAs), was proposed to valorize the organic fraction of municipal solid waste (OFMSW). Anaerobic digestion (AD) was tested at different organic loading rates (OLR-I-II-III) (i.e. 3, 4 and 6 g L-1 d-1, respectively), producing biomethane and volatile fatty acids (VFAs)-rich digestate, the VFAs were then used to produce PHAs. Specific biogas and CH4 production remained similar when adopting different OLRs (biogas of 522-600 NL kg-1 VS and CH4 of 64-67% v/v). VFAs concentrated with OLR increases and their patterns were modified. PHA production was in the range of 117-199 g kg-1 OFMSWTS with the lowest production being associated to different polymer composition. The net energy recovery of this simple biorefinery accounted for 64% of OFMSW energy content, and the PHAs produced represented over 30% of the total energy.

Production of ent-kaurene from lignocellulosic hydrolysate in Rhodosporidium toruloides.

BACKGROUND: Rhodosporidium toruloides has emerged as a promising host for the production of bioproducts from lignocellulose, in part due to its ability to grow on lignocellulosic feedstocks, tolerate growth inhibitors, and co-utilize sugars and lignin-derived monomers. Ent-kaurene derivatives have a diverse range of potential applications from therapeutics to novel resin-based materials. RESULTS: The Design, Build, Test, and Learn (DBTL) approach was employed to engineer production of the non-native diterpene ent-kaurene in R. toruloides. Following expression of kaurene synthase (KS) in R. toruloides in the first DBTL cycle, a key limitation appeared to be the availability of the diterpene precursor, geranylgeranyl diphosphate (GGPP). Further DBTL cycles were carried out to select an optimal GGPP synthase and to balance its expression with KS, requiring two of the strongest promoters in R. toruloides, ANT (adenine nucleotide translocase) and TEF1 (translational elongation factor 1) to drive expression of the KS from Gibberella fujikuroi and a mutant version of an FPP synthase from Gallus gallus that produces GGPP. Scale-up of cultivation in a 2 L bioreactor using a corn stover hydrolysate resulted in an ent-kaurene titer of 1.4 g/L. CONCLUSION: This study builds upon previous work demonstrating the potential of R. toruloides as a robust and versatile host for the production of both mono- and sesquiterpenes, and is the first demonstration of the production of a non-native diterpene in this organism.

Idiopathic thrombocytopenic purpura in a patient with situs inversus totalis: case report and literature review.

Situs inversus totalis is a rare recessive autosomal congenital abnormality in which the mediastinal and abdominal organs are in a mirrored position when compared to the usual topography. The literature reports some cases of situs inversus totalis and concomitant conditions: spinal abnormalities, cardiac malformations and hematological diseases, such as idiopathic thrombocytopenic purpura, which is an autoimmune disease that causes thrombocytopenia due to platelet destruction or suppression of its production. This article aimed to report the coexistence of situs inversus totalis and idiopathic thrombocytopenic purpura.

Idiopathic thrombocytopenic purpura in a patient with situs inversus totalis: case report and literature review / Púrpura trombocitopênica idiopática em paciente com situs inversus totalis: relato de caso e revisão da literatura

ABSTRACT Situs inversus totalis is a rare recessive autosomal congenital abnormality in which the mediastinal and abdominal organs are in a mirrored position when compared to the usual topography. The literature reports some cases of situs inversus totalis and concomitant conditions: spinal abnormalities, cardiac malformations and hematological diseases, such as idiopathic thrombocytopenic purpura, which is an autoimmune disease that causes thrombocytopenia due to platelet destruction or suppression of its production. This article aimed to report the coexistence of situs inversus totalis and idiopathic thrombocytopenic purpura.

RESUMO Situs inversus totalis é uma anormalidade congênita autossômica recessiva rara em que os órgãos mediastinais e abdominais encontram-se em posição espelhada em relação à topografia habitual. A literatura relata alguns casos de concomitância do situs inversus totalis com outras condições: anomalias espinhais, malformações cardíacas e doenças hematológicas, como púrpura trombocitopênica idiopática, que é uma doença autoimune com plaquetopenia, devido à destruição dos trombócitos ou supressão da sua produção. Esse artigo teve o objetivo de relatar coexistência de situs inversus totalis e púrpura trombocitopênica idiopática.

One-pot bio-derived ionic liquid conversion followed by hydrogenolysis reaction for biomass valorization: A promising approach affecting the morphology and quality of lignin of switchgrass and poplar.

The use of bio-derived ionic liquids (e.g., cholinium lysinate) in a one-pot process was evaluated on overall sugar and lignin yields as a function of two model woody and herbaceous feedstocks, switchgrass and poplar, with emphasis on the study of physical and chemical alterations in lignin structure, by performing a detailed mass balance analysis and chemical characterization. Multiple chromatographic and spectroscopic analytical techniques were applied tracking lignin reactivity and partitioning during the ionic liquid one-pot conversion. Depolymerization efficiency of the lignin-rich residue derived from the whole process was investigated as a function of different temperatures and pressures during catalytic hydrogenolysis by Ni(SO)4. This study validates the potential of ionic liquid one pot process as an integrated approach for full exploitation of lignocellulosic feedstocks. The insights gained will contribute to the design of future conversion routes for efficient biomass deconstruction and lignin valorization.

Parametric study for the optimization of ionic liquid pretreatment of corn stover.

A parametric study of the efficacy of the ionic liquid (IL) pretreatment (PT) of corn stover (CS) using 1-ethyl-3-methylimidazolium acetate ([C2C1Im][OAc]) and cholinium lysinate ([Ch][Lys]) was conducted. The impact of 50% and 15% biomass loading for milled and non-milled CS on IL-PT was evaluated, as well the impact of 20 and 5mg enzyme/g glucan on saccharification efficiency. The glucose and xylose released were generated from 32 conditions - 2 ionic liquids (ILs), 2 temperatures, 2 particle sizes (S), 2 solid loadings, and 2 enzyme loadings. Statistical analysis indicates that sugar yields were correlated with lignin and xylan removal and depends on the factors, where S did not explain variation in sugar yields. Both ILs were effective in pretreating large particle sized CS, without compromising sugar yields. The knowledge from material and energy balances is an essential step in directing optimization of sugar recovery at desirable process conditions.

Evaluation of Relationships between Growth Rate, Tree Size, Lignocellulose Composition, and Enzymatic Saccharification in Interspecific Corymbia Hybrids and Parental Taxa.

In order for a lignocellulosic bioenergy feedstock to be considered sustainable, it must possess a high rate of growth to supply biomass for conversion. Despite the desirability of a fast growth rate for industrial application, it is unclear what effect growth rate has on biomass composition or saccharification. We characterized Klason lignin, glucan, and xylan content with response to growth in Corymbia interspecific F1 hybrid families (HF) and parental species Corymbia torelliana and C. citriodora subspecies variegata and measured the effects on enzymatic hydrolysis from hydrothermally pretreated biomass. Analysis of biomass composition within Corymbia populations found similar amounts of Klason lignin content (19.7-21.3%) among parental and hybrid populations, whereas glucan content was clearly distinguished within C. citriodora subspecies variegata (52%) and HF148 (60%) as compared to other populations (28-38%). Multiple linear regression indicates that biomass composition is significantly impacted by tree size measured at the same age, with Klason lignin content increasing with diameter breast height (DBH) (+0.12% per cm DBH increase), and glucan and xylan typically decreasing per DBH cm increase (-0.7 and -0.3%, respectively). Polysaccharide content within C. citriodora subspecies variegata and HF-148 were not significantly affected by tree size. High-throughput enzymatic saccharification of hydrothermally pretreated biomass found significant differences among Corymbia populations for total glucose production from biomass, with parental Corymbia torelliana and hybrids HF-148 and HF-51 generating the highest amounts of glucose (~180 mg/g biomass, respectively), with HF-51 undergoing the most efficient glucan-to-glucose conversion (74%). Based on growth rate, biomass composition, and further optimization of enzymatic saccharification yield, high production Corymbia hybrid trees are potentially suitable for fast-rotation bioenergy or biomaterial production.

Phosphatase activity and its relationship with physical and chemical parameters during vermicomposting of filter cake and cattle manure.

BACKGROUND: Recycling of phosphorus (P) from organic residues (ORs) is important to develop environmentally friendly agriculture. The use of this P source depends on phosphatase enzymes, which can be affected by a chain of parameters during maturation of ORs. In this study the phosphatase activity levels throughout vermicomposting of filter cake (FC) and cattle manure (CM) were correlated with different physical and chemical parameters in an effort to increase the knowledge about recycling of P from ORs. RESULTS: FC presented higher total nitrogen content (TNC), total organic carbon (TOC), humic acid (HA) content, water-soluble P (WSP), phosphatase activities and nanopore volume than CM during vermicomposting. Decreases in TOC of CM resulted from carbohydrate mineralization, which was not observed for FC. CM showed increased hydrophobic index during vermicomposting while FC showed a slight decrease. CONCLUSION: Phosphatase activities correlated positively with TOC, pH and WSP and negatively with HA content for both vermicomposts. Nanopore volume was negatively correlated with phosphatase activities for FC but not for CM. No correlations between hydrophobicity and phosphatase activities were found for FC. Increased hydrophobicity throughout vermicomposting of CM could be partially associated with decreases in phosphatase levels.

Penetrance of Hemochromatosis in HFE Genotypes Resulting in p.Cys282Tyr and p.[Cys282Tyr];[His63Asp] in the eMERGE Network.

Hereditary hemochromatosis (HH) is a common autosomal-recessive disorder associated with pathogenic HFE variants, most commonly those resulting in p.Cys282Tyr and p.His63Asp. Recommendations on returning incidental findings of HFE variants in individuals undergoing genome-scale sequencing should be informed by penetrance estimates of HH in unselected samples. We used the eMERGE Network, a multicenter cohort with genotype data linked to electronic medical records, to estimate the diagnostic rate and clinical penetrance of HH in 98 individuals homozygous for the variant coding for HFE p.Cys282Tyr and 397 compound heterozygotes with variants resulting in p.[His63Asp];[Cys282Tyr]. The diagnostic rate of HH in males was 24.4% for p.Cys282Tyr homozygotes and 3.5% for compound heterozygotes (p < 0.001); in females, it was 14.0% for p.Cys282Tyr homozygotes and 2.3% for compound heterozygotes (p < 0.001). Only males showed differences across genotypes in transferrin saturation levels (100% of homozygotes versus 37.5% of compound heterozygotes with transferrin saturation > 50%; p = 0.003), serum ferritin levels (77.8% versus 33.3% with serum ferritin > 300 ng/ml; p = 0.006), and diabetes (44.7% versus 28.0%; p = 0.03). No differences were found in the prevalence of heart disease, arthritis, or liver disease, except for the rate of liver biopsy (10.9% versus 1.8% [p = 0.013] in males; 9.1% versus 2% [p = 0.035] in females). Given the higher rate of HH diagnosis than in prior studies, the high penetrance of iron overload, and the frequency of at-risk genotypes, in addition to other suggested actionable adult-onset genetic conditions, opportunistic screening should be considered for p.[Cys282Tyr];[Cys282Tyr] individuals with existing genomic data.

Perspective on the use of humic acids from biomass as natural surfactants for industrial applications.

In the context of renewable vs. non-renewable sources of chemical compounds, the development of natural surfactants as a substitute for synthetic surfactants in technological applications is an important issue. In addition, as synthetic surfactants can persist in the environment causing toxic effects, the use of natural products presents a possibility to minimize impact on the environment. Nowadays, a promising new approach in surfactant-based technologies, consists of the use of humic acids (HAs) extracted directly from biomass that exhibit amphiphilic properties, and can be conveniently used as environmentally friendly surfactants. The raw material from which HAs are extracted and their macromolecular composition affect surfactant properties. Therefore fundamental data from more strictly qualitative aspects, needs to be investigated. This review highlights surfactant ability and chemical properties of HA substances coming from renewable sources in comparison to synthetic surfactants, and points out the capacity for HAs to be used effectively in this field of application.

Nanoscale structure of the cell wall protecting cellulose from enzyme attack.

The cell wall structure protects cellulose from enzymatic attack and its successive fermentation. The nature of this protection consists in the very complex macroscopic and microscopic structure of cell wall that limits transport. Explaining this kind of protection is critical in future research to improve cell polymer availability for enzymatic attack. This research shows that the complete description of the cell wall topography at a nanoscale level allows a mechanistic understanding of cellulose protection. For this purpose, we used gas adsorption methods (CO(2) at 273 K and N(2) at 77 K) to detect mesoporosity (pore size of 1.5-30 nm diameter; MeS) and microporosity (pore size of 0.3-1.5 nm diameter; MiS) of the cell wall of five energy crops, i.e., giant cane, rivet wheat straw, miscanthus, proso millet, and sorghum. The presence of both hemicelluloses in the spaces between cellulose fibrils and the unhydrolyzable and highly cross-linked lignocarbohydrate complex (LCC) determines a microporous (80% pores having diameters below 0.8 nm) structure of the cell wall that prevents the cellulase enzymes from coming into direct contact with the cellulose, as their sizes exceed the cell wall pore size. On the other hand, the removal of the hemicelluloses and of the LCC complex determines a reduction of the MiS and an increase of the available surface for enzymatic attack, i.e., pores >5 nm diameter. This was confirmed by the good negative (r = -0.87, P < 0.001, n = 11) and positive (r = 0.78, P < 0.005, n = 11) correlations found for microporosity and mesoporosity (pores of diameters >5 nm), respectively, vs the glucose production, by cellulase enzyme attack in specific enzymatic hydrolysis tests performed on biomass samples.

Micropore surface area of alkali-soluble plant macromolecules (humic acids) drives their decomposition rates in soil.

Previous studies suggested that micropore surface area (MSA) of alkali-soluble bio-macromolecules of aerial plant residues of maize constitutes an important factor that explains their humification in soil, that is, preservation against biological degradation. On the other hand, root plant residue contributes to the soil humus balance, as well. Following the experimental design used in a previous paper published in this journal, this study shows that the biochemical recalcitrance of the alkali-soluble acid-insoluble fraction of the root plant material, contributed to the root maize humification of both Wild-type maize plants and its corresponding mutant brown midrib (bm3), this latter characterized by reduced lignin content. Humic acids (HAs) existed in root (root-HAs) were less degraded in soil than corresponding HAs existed in shoot (shoot-HAs): shoot-HAs bm3 (48%)>shoot-HAs Wild-type (37%)>root-HAs Wild-type (33%)>root-HAs bm3 (22%) (degradability shown in parenthesis). These differences were related to the MSA of HAs, that is, root-HAs having a higher MSA than shoot-HAs: shoot-HAs bm3 (41.43+/-1.2m(2)g(-1))