Solid state fermentation of straw withNeurospora crassa for CMCase and β-glucosidase production

Summary CMCase and -glucosidase were produced by the mutantNeurospora crassa 40b cultivated on untreated wheat straw in a solid state fermentation. Best enzyme activities were observed when the growth medium was composed of wheat straw mixed with certain mineral solutions at a ratio 1:2 (w/v). A partially purified enzyme preparation showed optimum enzyme activities of CMCase and -glucosidase at pH 4.0 and 5.0 and temperature 50 and 60°C respectively. The apparent Km values for the same enzymes were 16.8 g/l and 1.03x10^–4 M respectively. At optimum growth and enzyme assay conditions yields as high as 586.2 U CMCase and 58.4 U -glucosidase per gram of straw were obtained.     

Culturable and metagenomic approaches of wheat bran and wheat straw phyllosphere’s highlight new lignocellulolytic microorganisms

The phyllosphere, defined as the aerial parts of plants, is one of the most prevalent microbial habitats on earth. The microorganisms present on the phyllosphere can have several interactions with the plant. The phyllosphere represents then a unique niche where microorganisms have evolved through time in that stressful environment and may have acquired the ability to degrade lignocellulosic plant cell walls in order to survive to oligotrophic conditions. The dynamic lignocellulolytic potential of two phyllospheric microbial consortia (wheat straw and wheat bran) has been studied. The microbial diversity rapidly changed between the native phyllospheres and the final degrading microbial consortia after 48 h of culture. Indeed, the initial microbial consortia was dominated by the Ralstonia (35·8%) and Micrococcus (75·2%) genera for the wheat bran and wheat straw whereas they were dominated by Candidatus phytoplasma (59%) and Acinetobacter (31·8%) in the final degrading microbial consortia respectively. Culturable experiments leading to the isolation of several new lignocellulolytic isolates (belonging to Moraxella and Atlantibacter genera) and metagenomic reconstruction of the microbial consortia highlighted the existence of an unpredicted microbial diversity involved in lignocellulose fractionation but also the existence of new pathways in known genera (presence of CE2 for Acinetobacter, several AAs for Pseudomonas and several GHs for Bacillus in different metagenomes-assembled genomes). The phyllosphere from agricultural co-products represents then a new niche as a lignocellulolytic degrading ecosystem.     

Kinetics of wheat straw solid-state fermentation with Trametes versicolor and Pleurotus ostreatus — lignin and polysaccharide alteration and production of related enzymatic activities

Summary The kinetics of straw solid-state fermentation (SSF) with Trametes versicolor and Pleurotus ostreatus was investigated to characterize the delignification processes by these white-rot fungi. Two successive phases could be defined during straw transformation, characterized by changes in respiratory activity, changes in lignin and polysaccharide content and composition, increase in in-vitro digestibility, and enzymatic activities produced by the fungi. Lignin composition was analysed after CuO alkaline degradation, and decreases in syringyl/guaiacyl and syringyl/p-hydroxyphenyl ratios and cinnamic acid content were observed during the fungal treatment. An increase in the phenolic acid yield, revealing fungal degradation of side-chains in lignin, was produced by P. ostreatus. The highest xylanase level was produced by P. ostreatus, and exocellulase activity was nearly absent from straw treated with this fungus. Lactase activity was found in straw treated with both fungi, but lignin peroxidase was only detected during the initial phase of straw transformation with T. versicolor. High levels of H₂O₂-producing aryl-alcohol oxidase occurred throughout the straw SSF with P. ostreatus. Offprint requests to: A. T. Martínez     

Effect of endoxylanase and α-L-arabinofuranosidase supplementation on the enzymatic hydrolysis of steam exploded wheat straw

The cost and hydrolytic efficiency of enzymes are major factors that restrict the commercialization of the bioethanol production process from lignocellulosic biomass. Hemicellulases and other accessory enzymes are becoming crucial to increase enzymatic hydrolysis (EH) yields at low cellulase dosages. The aim of this work was to evaluate the effect of two recombinant hemicellulolytic enzymes on the EH of steam pretreated wheat straw. Pretreatments at two severity conditions were performed and the whole slurry obtained after steam explosion pretreatment was employed as substrate. An endoxylanase (Xln C) from Aspergillus nidulans and an α-l-arabinofuranosidase (AF) from Aspergillus niger, have been applied in combination with cellulase enzymes. A degree of synergism of 29.5% and increases up to 10% in the EH yields were obtained, showing the potential of accessory activities to improve the EH step and make the whole process more effective.     

Oxalate production at different initial Pb²+ concentrations and the influence of oxalate during solid-state fermentation of straw with Phanerochaete chrysosporium

The production of oxalate at different initial Pb²⁺ concentrations during solid-state fermentation of straw with Phanerochaete chrysosporium was investigated. It was found that the maximal peak value of oxalate concentration (22.84 mM) was detected at the initial Pb²⁺ concentration of 200 mg kg⁻¹ dry straw, while the minimum (15.89 mM) at the concentration of 600 mg Pb²⁺ kg⁻¹ dry straw, and at moderate concentration of Pb²⁺ the capability of oxalic acid secretion was enhanced. In addition, it was also found that more oxalic acid accumulation went together with better Pb²⁺ passivation effect and higher manganese peroxidase (MnP) activity. The present findings will improve the understandings of the interactions of heavy metals with white-rot fungi and the role of oxalate in lignin degradation system, which could provide useful references for more efficient treatment of Pb-contaminated lignocellulosic waste.     

Immobilized β-glucosidase on magnetic chitosan microspheres for hydrolysis of straw cellulose

β-Glucosidase immobilized on magnetic chitosan microspheres for potential recycling usage in hydrolysis of cellulosic biomass was investigated. The immobilized enzyme had an activity of 6.4 U/g support under optimized condition when using cellobiose as substrate. Immobilization resulted in less increase of the apparent K_m, low drift of the optimal pH, as well as improved stability relative to the free enzyme. The immobilized β-glucosidase was applied to enzymatic hydrolysis of corn straw to produce 60.2 g/l reducing sugar with a conversion rate of 78.2% over the course of a 32-h reaction. This conversion rate was maintained above 76.5% after recycling the enzyme for use in eight batches (total 256 h), showing favorable operational stability of the immobilized enzyme.     

Effect of tillage and straw return on carbon footprints,soil organic carbon fractions and soil microbial community in different textured soils under rice–wheat rotation: a review

Reviews in Environmental Science and Bio/Technology - Measuring the influence of long-term agricultural tillage practices on soil organic carbon (SOC) is of great importance to farmers and...     

Degradation of wheat straw by a microbial community —stimulation by a polysaccharidase complex

Laboratory-scale reactors were used to watch aspects of biodegradation of wheat straw when supplemented with polysaccharidases (Czym) to increase the enzyme production of microorganisms involved during a composting process for mushroom production. Biochemical and biological parameters were tested both under aerobic and O₂-limited conditions to assess degradability. These were measurement of released CO₂ and NH₃, determination of neutral detergent fibre content and cellulase activities from compost extract. The addition of Czym to decomposing straw had three consequences: (i) it supplied and released low quantities of readily available sugars; (ii) it increased the cellulase activities in the substrates; (iii) it increased the number of bacteria under aerobic conditions. The three effects were linked and the small quantity of sugars released by the addition of Czym may have acted as an activator of bacterial activities through an inductive mechanism. Correspondence to: S. Libmond     

Properties important for solid–liquid separations change during the enzymatic hydrolysis of pretreated wheat straw

Objectives

The biochemical conversion of lignocellulosic biomass into renewable fuels and chemicals provides new challenges for industrial scale processes. One such process, which has received little attention, but is of great importance for efficient product recovery, is solid–liquid separations, which may occur both after pretreatment and after the enzymatic hydrolysis steps. Due to the changing nature of the solid biomass during processing, the solid–liquid separation properties of the biomass can also change. The objective of this study was to show the effect of enzymatic hydrolysis of cellulose upon the water retention properties of pretreated biomass over the course of the hydrolysis reaction.

Results

Water retention value measurements, coupled with ¹H NMR T₂ relaxometry data, showed an increase in water retention and constraint of water by the biomass with increasing levels of cellulose hydrolysis. This correlated with an increase in the fines fraction and a decrease in particle size, suggesting that structural decomposition rather than changes in chemical composition was the most dominant characteristic.

Conclusions

With increased water retained by the insoluble fraction as cellulose hydrolysis proceeds, it may prove more difficult to efficiently separate hydrolysis residues from the liquid fraction with improved hydrolysis.

     

Improving grain yield in backcross populations from Avena sativa × A. sterilis matings by using independent culling for harvest index and vegetative growth index or unit straw weight

Summary Grain yield was selected in Avena sativa X A. sterilis populations of segregates by applying independent culling for harvest index and either vegetative growth index or unit straw weight. Samples of lines intensively selected for harvest index had high harvest index but low grain yield because they had low vigor. Populations intensively selected for growth rate index or unit straw weight had high biological yield but low harvest index. Intensive selection for grain yield per se resulted in samples with high grain yield, but they were late and tall. Samples selected for harvest index at a 25% selection intensity first and subsequently selected for vegetative growth index or unit straw weight had grain yield as high as the samples selected for grain yield per se and vegetative growth index, and they had acceptable heading date and plant height. Backcrosses three and four were best, among the various BC generations, for selecting oat lines with high grain yield and suitable agronomic traits. CI 7463 was superior to CI 8044 as a recurrent parent, and B 445 was inferior to other A. sterilis accessions as a donor parent.Journal Paper No. J-12287 of the Iowa Agric, and Home Econ. Exp. Stn., Ames, IA 50011. Project 2447. This research was supported in part by a grant from the World Food Institute, Iowa State University, Ames, IA, USA     

Methane emissions from two breeds of beef cows offered diets containing barley straw with either grass silage or brewers’ grains

Increasing the concentration of dietary lipid is a promising strategy for reducing methane (CH₄) emissions from ruminants. This study investigated the effect of replacing grass silage with brewers’ grains on CH₄ emissions of pregnant, non-lactating beef cows of two breeds. The experiment was a two×two factorial design comprising two breeds (LIMx, crossbred Limousin; and LUI, purebred Luing) and two diets consisting of (g/kg diet dry matter (DM)) barley straw (687) and grass silage (301, GS), or barley straw (763) and brewers’ grains (226, BG), which were offered ad libitum. Replacing GS with BG increased the acid-hydrolysed ether extract concentration from 21 to 37 g/kg diet DM. Cows (n=48) were group-housed in equal numbers of each breed across two pens and each diet was allocated to one pen. Before measurements of CH₄, individual dry matter intake (DMI), weekly BW and weekly body condition score were measured for a minimum of 3 weeks, following a 4-week period to acclimatise to the diets. CH₄ emissions were subsequently measured on one occasion from each cow using individual respiration chambers. Due to occasional equipment failures, CH₄ measurements were run over 9 weeks giving 10 observations for each breed×treatment combination (total n=40). There were no differences between diets for daily DMI measured in the chambers (9.92 v. 9.86 kg/day for BG and GS, respectively; P>0.05). Cows offered the BG diet produced less daily CH₄ than GS-fed cows (131 v. 156 g/day: P<0.01). When expressed either as g/kg DMI or kJ/MJ gross energy intake (GEI), BG-fed cows produced less CH₄ than GS-fed cows (13.5 v. 16.4 g/kg DMI, P<0.05; 39.2 v. 48.6 kJ/MJ GEI, P<0.01). Breed did not affect daily DMI or CH₄ expressed as g/day, g/kg DMI or kJ/MJ GEI (P>0.05). However, when expressed as a proportion of metabolic BW (BW^0.75), LUI cows had greater DMI than LIMx cows (84.5 v. 75.7 g DMI/kg BW^0.75, P<0.05) and produced more CH₄ per kg BW^0.75 than LIMx cows (1.30 v. 1.05 g CH₄/kg BW^0.75; P<0.01). Molar proportions of acetate were higher (P<0.001) and propionate and butyrate lower (P<0.01) in rumen fluid samples from BG-fed compared with GS-fed cows. This study demonstrated that replacing GS with BG in barley straw-based diets can effectively reduce CH₄ emissions from beef cows, with no suppression of DMI.     

Natural variation in the glucose content of dilute sulfuric acid–pretreated rice straw liquid hydrolysates: implications for bioethanol production

Rice straw is a promising resource for bioethanol production. Because the glucose content of pretreatment liquid hydrolysates is highly correlated with ethanol yield, the selection of appropriate rice cultivars is essential. The glucose content in liquid hydrolysates of pretreated rice straws of 208 diverse cultivars was evaluated in natural field in 2013 and 2014 using a novel high-throughput system. The glucose content of the rice straw samples varied across cultivars and was affected by environmental factors such as temperature and solar radiation. Several high-quality cultivars exhibiting high glucose content in both years were identified. The results of this study can aid in development of novel rice cultivars suitable as both feedstocks for bioethanol production and cooking.     

Production of β-xylosidase from <Emphasis Type="Italic">Trichoderma asperellum</Emphasis> KIF125 and its application in efficient hydrolysis of pretreated rice straw with fungal cellulase

On-site cellulase and hemicellulase production is a promising way to reduce enzyme cost in the commercialization of the lignocellulose-to-ethanol process. A hemicellulase-producing fungal strain suitable for on-site enzyme production was selected from cultures prepared using wet disc-milling rice straw (WDM-RS) and identified as Trichoderma asperellum KIF125. KIF125 hemicellulase showed uniquely high abundance of β-xylosidase in the xylanolytic enzyme system compared to other fungal hemicellulase preparations. Supplementation of Talaromyces cellulolyticus cellulase with KIF125 hemicellulase was more effective than that with the hemicellulases from other fungal sources in reducing the total enzyme loading for the improvement of xylose yield in the hydrolysis of ball-milling RS, due to its high β-xylosidase dominance. β-Xylosidase in KIF125 hemicellulase was purified and classified as a glycosyl hydrolase family 3 enzyme with relatively high specificity for xylobiose. The production of KIF125 β-xylosidase in the fermentor was estimated as 118 U/g-WDM-RS (2350 U/L culture) at 48 h. These results demonstrate that KIF125 is promising as a practical hemicellulase source to combine with on-site cellulase production using T. cellulolyticus.     

Which is the most preventive measure against tail damage in finisher pigs: tail docking,straw provision or lowered stocking density?

One challenge of intensive pig production is tail damage caused by tail biting, and farmers often decrease the prevalence of tail damage through tail docking. However, tail docking is not an optimal preventive measure against tail damage and thus, it would be preferable to replace it. The aim of the current study was to investigate the relative effect of three possible preventive measures against tail damage. The study included 112 pens with 1624 finisher pigs divided between four batches. Pens were randomly assigned to one level of each of three treatments: (1) tail-docked (n=60 pens) v. undocked (n=52 pens), (2) 150 g of straw provided per pig per day on the solid floor (n=56 pens) v. no straw provided (n=56 pens), (3) stocking density of 1.21 m²/pig (11 pig/pen; n=56 pens) v. 0.73 m²/pig (18 pigs/pen; n=56 pens). Tail damage was recorded three times per week throughout the finisher period by scoring the tail of each individual pig. A pen was recorded as a tail damage pen and no longer included in the study if at least one pig in a pen had a bleeding tail wound; thus, only the first incidence of tail damage on pen level was recorded. Data were analysed by a Cox regression for survival analysis assuming proportional hazards. Results are presented as hazards, and a higher hazard means that a pen has a higher risk of tail damage and of it happening earlier in the finisher period. Pens with undocked pigs had a 4.32-fold higher hazard of tail damage compared with pens with docked pigs (P<0.001). Pens with no straw provided had a 2.22-fold higher hazard of tail damage compared with pens with straw provided (P<0.01). No interactions was seen between the treatments, but the effect of tail docking was higher than the effect of straw provision (P<0.001). Stocking density did not have a significant effect on the hazard of tail damage (hazard rate ratios (HRR)=1.67; P=0.064). However, a combination of straw provision and lowered stocking density showed a similar hazard of tail damage as seen with only tail docking (HRR=1.58; P=0.39). In conclusion, tail docking and straw provision were preventive measures against tail damage, and tail docking reduced the risk more than straw provision. A combination of other preventive measures is necessary to reduce the risk of tail damage in undocked pigs to the same level as in docked pigs.     

High level expression of Acidothermus cellulolyticus β-1, 4-endoglucanase in transgenic rice enhances the hydrolysis of its straw by cultured cow gastric fluid

Background

In the hydrolysis of lignocellulosic materials, thermostable enzymes decrease the amount of enzyme needed due to higher specific activity and elongate the hydrolysis time due to improved stability. For cost-efficient use of enzymes in large-scale industrial applications, high-level expression of enzymes in recombinant hosts is usually a prerequisite. The main aim of the present study was to compare the biochemical and hydrolytic properties of two thermostable recombinant glycosyl hydrolase families 10 and 11 (GH10 and GH11, respectively) xylanases with respect to their potential application in the hydrolysis of lignocellulosic substrates.

Results

The xylanases from Nonomuraea flexuosa (Nf Xyn11A) and from Thermoascus aurantiacus (Ta Xyn10A) were purified by heat treatment and gel permeation chromatography. Ta Xyn10A exhibited higher hydrolytic efficiency than Nf Xyn11A toward birchwood glucuronoxylan, insoluble oat spelt arabinoxylan and hydrothermally pretreated wheat straw, and it produced more reducing sugars. Oligosaccharides from xylobiose to xylopentaose as well as higher degree of polymerization (DP) xylooligosaccharides (XOSs), but not xylose, were released during the initial hydrolysis of xylans by Nf Xyn11A, indicating its potential for the production of XOS. The mode of action of Nf Xyn11A and Ta Xyn10A on glucuronoxylan and arabinoxylan showed typical production patterns of endoxylanases belonging to GH11 and GH10, respectively.

Conclusions

Because of its high catalytic activity and good thermostability, T. aurantiacus xylanase shows great potential for applications aimed at total hydrolysis of lignocellulosic materials for platform sugars, whereas N. flexuosa xylanase shows more significant potential for the production of XOSs.     

Syntheses and spectroscopic properties of α- and β-phosphatidylcholines and phosphatidylethanolamines

The widely used partial synthesis of phospholipids via deacylation of naturally occurring phospholipids, followed by reacylation with fatty acid anhydrides, is accompanied by phosphoryl migration. The resulting mixture of α- and β-phospholipids was separated by short-column chromatography. Milder acylation procedures in which no phosphoryl migration occurs, were developed. 1,2-Dilinoleoyl-sn-glycero-3-phosphocholine was prepared in 50% yield by acylation of sn-glycero-3-phosphocholine (GPC) with N-linoleoylimidazole. Detailed NMR and infrared spectra of α- and β-phosphatidylcholines (PCs) and -ethanolamines (PEs) are reported and the differences between isomers discussed.     

Phenotypic plasticity and longevity in plants and animals: cause and effect?

Immobile plants and immobile modular animals outlive unitary animals. This paper discusses competing but not necessarily mutually exclusive theories to explain this extreme longevity, especially from the perspective of phenotypic plasticity. Stem cell immortality, vascular autonomy, and epicormic branching are some important features of the phenotypic plasticity of plants that contribute to their longevity. Monocarpy versus polycarpy can also influence the kind of senescent processes experienced by plants. How density-dependent phenomena affecting the establishment of juveniles in these immobile organisms can influence the evolution of senescence, and consequently longevity, is reviewed and discussed. Whether climate change scenarios will favour long-lived or short-lived organisms, with their attendant levels of plasticity, is also presented.     

Dogmatisms and Catechisms — Ethics and Companion Animals

Abstract

The current reconsideration of the place in nature of human beings unfortunately continues to be an acrimonious one. All too often the debate is more akin to a warlike encounter where each side attempts to gain control or the upper hand than a search for points of agreement. Given this context, it is important to entertain views that emanate from different cultural traditions as a way to infuse the debate with new life. Students of Native American culture have consistently pointed out that the essential concepts of life balance and reciprocity represented there may serve as useful points of consideration as we struggle with the appropriate relationships with animals and nature. This article presents a representative Zuni story, told by Governor Robert E. Lewis, that illustrates these notions.