Effects of cellulase treatment on properties of lignocellulose-based biochar.

As the most abundant renewable carbon source, lignocellulose holds potential as a raw material for biofuels and biochar. The components required for biofuel production differ from those for biochar, so combining processes can reduce costs. Biofuel preparation necessitates cellulase treatment of lignocellulose. This study examines the effects of various enzyme treatment conditions (dosage, time, temperature) on lignocellulose, focusing on the properties of biochar derived from it (BC-SR). A mathematical model was constructed to study the relationship between enzyme treatment conditions and BC-SR properties. BC-SR exhibited high adsorption selectivity for bisphenol A and outperformed untreated biochar in fixed-bed column experiments, demonstrating greater removal efficiency and structural integrity. This study provides insights into the impact of enzymatic treatment on biochar and offers a cost-effective method for producing stable, efficient biochar. Additionally, a highly persistent biochar can enter the carbon trading market as a carbon-neutral technology, further realizing economic and environmental benefits.

Biochar promotes microbial CO2 fixation by regulating feedback inhibition of metabolites.

Chemoautotrophs, the crucial contributors to biological carbon fixation, derive energy from reducing specific inorganic substances and utilize CO2 for growth. However, the release of extracellular free organic carbon (EFOC) by chemoautotrophic microorganisms can inhibit their own growth and metabolism. To reduce the feedback inhibition effect, a low-release biochar (BC-LR) was applied to adsorb EFOC. BC-LR not only adsorbed EFOC, but also selectively adsorbed the main inhibitory component, low molecular weight organics, in EFOC. In contrast, ordinary biochar could not effectively adsorb EFOC and its addition inhibited microbial growth and CO2 fixation. In Transwell culture, BC-LR promoted microbial growth by 190% and CO2 fixation by 29%, and exhibited better economic advantage, when compared with granular activated carbon. These findings provide a novel insight into the interaction between biochar and autotrophic microbial metabolism, offering an economically feasible approach to mitigate feedback inhibition of metabolites and promoting biological CO2 fixation.

Nanobubbles improve peroxymonosulfate-based advanced oxidation: High efficiency, low toxicity/cost, and novel collaborative mechanism.

Cl- activated peroxymonosulfate (PMS) oxidation technology can effectively degrade pollutants, but the generation of chlorinated disinfection byproducts (DBPs) limits the application of this technology in water treatment. In this study, a method of nanobubbles (NBs) synergistic Cl-/PMS system was designed to try to improve this technology. The results showed the synergistic effects of NBs/Cl-/PMS were significant and universal while its upgrade rate was from 12.89% to 34.97%. Moreover, the synergistic effects can be further improved by increasing the concentration and Zeta potential of NBs. The main synergistic effects of NBs/Cl-/PMS system were due to the electrostatic attraction of negatively charged NBs to Na+ from NaCl, K+ from PMS, and H+ from phenol, which acted as a "bridge" between Cl- and HSO5- as well as phenol and Cl-/HSO5-, increasing active substance concentration. In addition, the addition of NBs completely changed the oxidation system of Cl-/PMS from one that increases environmental toxicity to one that reduces it. The reason was that the electrostatic attraction of NBs changed the active sites and degradation pathway of phenol, greatly reducing the production of highly toxic DBPs. This study developed a novel environmentally friendly oxidation technology, which provides an effective strategy to reduce the generation of DBPs in the Cl-/PMS system.

A novel mycelial pellet applied to remove polycyclic aromatic hydrocarbons: High adsorption performance & its mechanisms.

Mycelial pellets formed by Penicillium thomii ZJJ were applied as efficient biosorbents for the removal of polycyclic aromatic hydrocarbons (PAHs), which are a type of ubiquitous harmful hydrophobic pollutants. The live mycelial pellets were able to remove 93.48 % of pyrene at a concentration of 100 mg/L within 48 h, demonstrating a maximum adsorption capacity of 285.63 mg/g. Meanwhile, the heat-killed one also achieved a removal rate of 65.01 %. Among the six typical PAHs (pyrene, phenanthrene, fluorene, anthracene, fluoranthene, benzo[a]pyrene), the mycelial pellets preferentially adsorbed the high molecular weight PAHs, which also have higher toxicity, resulting in higher removal efficiency. The experimental results showed that the biosorption of mycelial pellets was mainly a spontaneous physical adsorption process that occurred as a monolayer on a homogeneous surface, with mass transfer being the key rate-limiting step. The main adsorption sites on the surface of mycelia were carboxyl and N-containing groups. Extracellular polymeric substances (EPS) produced by mycelial pellets could enhance adsorption, and its coupling with dead mycelia could achieve basically the same removal effect to that of living one. It can be concluded that biosorption by mycelial pellets occurred due to the influence of electrostatic and hydrophobic interactions, consisting of five steps. Furthermore, the potential applicability of mycelial pellets has been investigated considering diverse factors. The mycelia showed high environmental tolerance, which could effectively remove pyrene across a wide range of pH and salt concentration. And pellets diameters and humic acid concentration had a significant effect on microbial adsorption effect. Based on a cost-effectiveness analysis, mycelium pellets were found to be a low-cost adsorbent. The research outcomes facilitate a thorough comprehension of the adsorption process of pyrene by mycelial pellets and their relevant applications, proposing a cost-effective method without potential environmental issues (heat-killed mycelial pellets plus EPS) to removal PAHs.

Effects of functional group loss on biochar activated persulfate in-situ remediation of phenol pollution in groundwater and its countermeasures.

Biochar is considered a good activator for use in advanced oxidation technology. However, dissolved solids (DS) released from biochar cause unstable activation efficiency. Biochar prepared from saccharification residue of barley straw (BC-SR) had less DS than that prepared directly from barley straw (BC-O). Moreover, BC-SR had a higher C content, degree of aromatization, and electrical conductivity than BC-O. Although the effects of BC-O and BC-SR on activation of Persulfate (PS) to remove phenol were similar, the activation effect of DS from BC-O was 73% higher than that of DS from BC-SR. Moreover, the activation effect of DS was shown to originate from its functional groups. Importantly, BC-SR had higher activation stability than BC-O owing to the stable graphitized carbon structure of BC-SR. Identification of reactive oxygen species showed that SO4•-, •OH, and 1O2 were all effective in degradation by BC-SR/PS and BC-O/PS systems, but their relative contributions differed. Furthermore, BC-SR as an activator showed high anti-interference ability in the complex groundwater matrix, indicating it has practical application value. Overall, this study provides novel insight that can facilitate the design and optimization of a green, economical, stable, and efficient biochar-activated PS for groundwater organic pollution remediation.

Influence of Different Pretreatment Steps on the Ratio of Phenolic Compounds to Saccharides in Soluble Polysaccharides Derived from Rice Straw and Their Effect on Ethanol Fermentation.

The complex structure of rice straw is such that its bioconversion requires multiple physical and chemical pretreatment steps. In this study, it was found that a large amount of soluble polysaccharides (SPs) are formed during the pretreatment of straw. The yield of NaOH-based SPs (4.8%) was much larger than that of ball-milled SPs (1.5%) and H2SO4-based SPs (1.1%). For all the pretreatments, the ratio of phenolic compounds to saccharides (P/S) for each type of SPs increased upon increasing the concentration of ethanol in the order of 90% > 70% > 50%. The yield of NaOH-based SPs was much higher than that of acid-based and ball-milled SPs. The changes in the 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS), ferric reducing antioxidant power assay (FRAP), and 2,2-diphenyl-1-picrylhydrazyl (DPPH) of SPs follow the same rule, i.e., the higher the P/S ratio, the higher the antioxidant values of the SPs. The flow cytometry and laser scanning microscopy results show that the P/S ratio can significantly influence the effect of SPs on microbial growth and cell membrane permeability. Upon varying the ethanol concentration in the range of 50-90%, the P/S ratio increased from 0.02 to 0.17, resulting in an increase in the promoting effects of the SPs on yeast cell growth. Furthermore, H2O2, NAD+/NADH, and NADP+/NADPH assays indicate that SPs with a high P/S ratio can reduce intracellular H2O2 and change the intracellular redox status.

Antibacterial Effect of Phenolic Acids Derived from Rice Straw and in Combination with Antibiotics Against Escherichia coli.

Many studies have demonstrated that natural plant extracts have inhibitory effects on microorganisms. The purpose of this study was to investigate the inhibitory effect of phenolic acids from rice straw (PAs) on Escherichia coli and their synergistic effect in combination with antibiotics. PAs can inhibit the growth of E. coli effectively by inducing the formation of H2O2; PA-treated cells had a tenfold greater intracellular H2O2 concentration than the control group. The synergistic effect caused by the interaction of PAs and antibiotics on inhibiting the growth of E. coli was significant. This effect may be caused by a PA-induced change in the permeability of E. coli cell membrane. The treatment with PAs made the extracellular K+ concentration reached 15 mg/L within 30 min, while the K+ concentration in the control group was very low and did not change significantly over time. Similarly to the extracellular K+, the extracellular protein concentration exceeded 150 mg/L in the PA treatment group, while it remained very low in the control group. Due to the increased cell permeability, more antibiotics can enter the cell. Hence, this study may provide a novel method of improving the safe use of antibiotics.

Effect of dissolved solids released from biochar on soil microbial metabolism.

Dissolved solids released from biochar (DSRB), including organic and inorganic compounds, may affect the role of biochar as a soil amendment. In this study, the effects of DSRB on soil microbe metabolism, especially CO2 fixation, were evaluated in liquid soil extract. DSRB were found to be released in large amounts (289.05 mg L-1 at 1 hour) from biochar over a short period of time before their rate of release slowed to a gradual pace. They increased the microbial biomass and provided energy and reducing power to microbes, while reducing their metabolic output of extracellular proteins and polysaccharides. DSRB inputs led to the redistribution of metabolic flux in soil microorganisms and an increased organic carbon content in the short term. This content gradually decreased as it was utilized. DSRB did not improve microbial CO2 fixation but, rather, enhanced its release, while promoting specific soil microorganism genera, including Cupriavidus, Flavisolibacter, and Pseudoxanthomonas. These heterotrophic genera may compete with autotrophic microorganisms for nutrients but have positive synergistic relationships with autotrophs during CO2 fixation. These results demonstrated that reducing the DSRB in biochar can improve its role as a soil amendment by enhancing soil carbon storage and CO2 fixation capabilities.

Oligosaccharides in straw hydrolysate could improve the production of single-cell protein with Saccharomyces cerevisiae.

BACKGROUND: Using agricultural wastes to produce single-cell proteins (SCP) can reduce production costs effectively. The aims of this study were to investigate the effects of enzyme loading on the components of rice straw (RS) hydrolysate and their effects on the growth of yeast. RESULTS: At the same glucose concentration, the dry weight of cells produced in the hydrolysate was 2.89 times higher than that in 2 g L-1 yeast extract (YE) medium, indicating that the hydrolysate was a suitable substrate for yeast growth. Ethanol precipitation followed by analysis showed that there were many oligosaccharides in the hydrolysate. The amount of cellulase had an important effect on the production of monosaccharides but had a smaller effect on the amounts and compositions of oligosaccharides. Adding oligosaccharides to the medium had no effect on ethanol production, but it promoted yeast growth and increased SCP production effectively. The results indicate that oligosaccharides were an important growth factor for yeast in the hydrolysate. Compared with YE medium, the cost of the medium with the hydrolysate was reduced by 68.47% when the same dry cell weight was obtained. CONCLUSION: Oligosaccharides in the hydrolysate can improve SCP production with low nutrient cost. This finding could reduce the amounts of cellulase required during saccharification and nutrients during culture, providing a new low-cost method for SCP production. © 2021 Society of Chemical Industry.

Utilization of straw-based phenolic acids as a biofugicide for a green agricultural production.

Phenolic compounds inhibit phytopathogenic fungal infections effectively. In this study, the antifungal effects of rice straw-derived phenolic acids (PAs) against Fusarium oxysporum were investigated. PAs can inhibit hyphal growth and spore germination, and p-coumaric acid (CA) is the main antifungal substance in PAs. PAs could induce the formation of hydrogen peroxide and increase the relative conductivity and extracellular K+ concentration. Observations using Scanning Electron Microscopy, Laser Scanning Confocal Microscopy and Transmission Electron Microscopy revealed that PAs could damage membrane permeability, which caused cytoplasm leakage. This phenomenon was verified by conductivity and the release of extracellular K+. The chlorophyll fluorescence maps of tomato leaves suggested that F. oxysporum damaged the tomato' photosynthetic system and that PAs reduced the area infected, thereby alleviating the damage. Moreover, PAs could decrease the disease incidence of tomato fruit. The results confirmed the feasibility of using PAs as a biofungicide and provide a way to increase the value of rice straw.

Reduction of Fermentation-Associated Stresses by Straw-Based Soluble Saccharides for Enhancing Ethanol Production.

In this study, the effect of soluble polysaccharides (SPs) derived from agricultural waste, rice straw, on fermentation-associated stresses (temperature and concentrations of glucose and ethanol) was investigated to achieve high-performance ethanol production. The increase in temperature and concentrations of glucose and ethanol significantly inhibited Saccharomyces cerevisiae growth and lowered ethanol fermentation efficiency. Flow cytometric assays indicated that SPs could alleviate membrane permeability damage caused by fermentation-associated stresses. Atomic force microscopy and transmission electron microscopy revealed that fermentation-associated stresses induced cell surface shrinkage, causing a decrease in the cell size, whereas SPs stimulated the formation of extracellular matrices (EMs), which made the cell surface smooth and the cell morphology regular. Cells with EMs induced by SPs could efficiently produce ethanol under severe stresses. As a result, the titer of ethanol in the fermentation with SPs was 1.40-fold (from 26.40 to 36.98 g/L) higher than that in the fermentation without SPs, suggesting the stress-alleviating effect of SPs on ethanol production.

Simultaneous production of cellulase and ferulic acid esterase by Penicillium decumbens with rice straw as the sole carbon source.

As well as cellulose and hemicelluloses, rice straw contains phenolic acids. The simultaneous production of monosaccharides and phenolic acids could improve the value of rice straw. In this study, it was confirmed that Penicillium decumbens produces more ferulic acid esterase (FAE) than other cellulase-producing fungi. Cellulose, destarched wheat bran (DSWB), and rice straw were used as carbon sources. Little phenolic acid was released by cellulose- and DSWB-based enzymes during the saccharification of rice straw, whereas rice straw was a favorable carbon source for the simultaneous production of cellulase and FAE. High-performance liquid chromatography showed that during enzyme production, phenolic acids were released from rice straw, and ball-milling affected this release of phenolic acids. Small amounts of phenolic acids induced FAE production. Although the enzymes produced with rice straw showed lower FAE activity than those produced with DSWB, phenolic acids were produced efficiently during the saccharification of rice straw in response to the synergistic effects of cellulase and FAE. Therefore, we suggest that the production of enzymes by P. decumbens on rice straw as the sole carbon source will allow the production of more valuable products from rice straw, making the utilization of rice straw more economic.

Saccharides in straw hydrolysate decrease cell membrane damage by phenolics by inducing the formation of extracellular matrix in yeast.

The bioconversion of rice straw into ethanol can alleviate the energy crisis and solve problems related to waste treatment. In this study, the effect of soluble polysaccharides (SPs) produced during rice straw saccharification on the formation of extracellular matrices (EMs) by the yeast Saccharomyces cerevisiae was investigated. SPs were characterized by high-performance liquid chromatography (HPLC) and fourier transform infrared spectroscopy (FT-IR). SPs reduced the inhibition of alcohol dehydrogenase activity by phenolic acids (PAs) and regulated the intracellular redox state, resulting in higher ethanol production. The results of flow cytometry, confocal laser scanning microscopy, and atomic force microscopy indicated that PAs changed microbial morphology and caused damage in microbial cell membranes. The protective effect of SPs against cell membrane damage could be attributed to the synthesis of polysaccharide-dependent extracellular matrix, which maintained cellular integrity even under phenolic acid stress. These findings provide new strategies to improve pretreatment and saccharification processes.

Extraction of Flavonoids from the Saccharification of Rice Straw Is an Integrated Process for Straw Utilization.

To date, bioethanol is not economically competitive. One strategy to overcome this limitation is co-producing ethanol and high value-added products as an integrated process. The results of this study demonstrated that flavonoids could be extracted from rice straw, and the flavonoids apigenin and kaempferol were detected by HPLC. Compared with untreated straw, ball-milling slightly increased the total amount of flavonoids and antioxidant activity measured by ABTS, DPPH, and FRAP assays. The saccharification step in the bioconversion of straw strongly affected the extraction of flavonoids from straw. The residue obtained after saccharification of ball-milled straw for glucose production was more suitable for flavonoid extraction than untreated and ball-milled straw. The yield of flavonoids from the residue was 1.51-fold higher than that from untreated straw. The antioxidant activity of flavonoids derived from the residue was similar to that of flavonoid-rich biomasses such as rice bran and wheat bran. More importantly, saccharification may significantly affect the conditions of flavonoid extraction. In this respect, treatment with cellulase may reduce the extraction time from 2.0 to 0.5 h and the extraction temperature from 80 to 30 °C. Therefore, saccharification in the bioconversion of straw may be considered as an enzyme pretreatment step for the efficient extraction of flavonoids from straw, serving as a sustainable process for straw utilization.

Utilization of the saccharification residue of rice straw in the preparation of biochar is a novel strategy for reducing CO2 emissions.

Once rice straw has been bioconverted into biofuels, it is difficult to further biodegrade or decompose the saccharification residue (mainly lignin). Taking into account the pyrolysis characteristics of lignin, in this study the saccharification residue was used as a raw material for the preparation of biochar (biochar-SR), a potential soil amendment. Biochar was prepared directly from rice straw (biochar-O) with a yield of 32.45 g/100 g rice straw, whereas 30.14 g biochar-SR and 30.46 g monosaccharides (including 20.46 g glucose, 9.11 g xylose, and 0.89 g arabinose) were obtained from 100 g of rice straw. When added to liquid soil extracts as a soil amendment, almost nothing was released from biochar-SR, whereas numerous dissolved solids (about 70 mg/L) were released from biochar-O. Adding a mixture of biochar-SR and autotrophic bacteria improved soil total organic carbon 1.8-fold and increased the transcription levels of cbbL and cbbM, which were 4.76 × 103 and 3.76 × 105 times those of the initial blank, respectively. By analyzing the soil microbial community, it was clear that the above mixture favored the growth of CO2-fixing bacteria such as Ochrobactrum. Compared with burning rice straw or preparing biochar-O, the preparation of biochar-SR reduced CO2 emissions by 67.53% or 37.13%, respectively. These results demonstrate that biochar-SR has potential applications in reducing the cost of sustainable energy and addressing environmental issues.

Tapping the Bioactivity Potential of Residual Stream from Its Pretreatments May Be a Green Strategy for Low-Cost Bioconversion of Rice Straw.

In this study, it was found that the residual stream from pretreatments of rice straw exhibited high antioxidant activity. Assays based on the Folin-Ciocalteu colorimetric method confirmed that the residual stream contained large amounts of phenolic compounds. Three antioxidant assays were employed to evaluate the bioactivity of the residual stream. Strong linear correlations existed among the release of phenolic compounds, saccharification efficiency, and antioxidant activity. The alkaline pretreatment provided a much greater release of phenolic compounds, especially phenolic acids, compared to the acid pretreatment, and consequently, it had stronger linear correlations than the acid pretreatment. Antibacterial experiments demonstrated the ability of the phenolic compounds in the residual stream to inhibit the growth of microorganisms, indicating the potential of these compounds as antimicrobial agents. To discuss the possibility of the co-production of antimicrobial agents and biofuels/biochemicals, both acid and alkaline pretreatments were optimized using response surface methodology. Under the optimal conditions, 285.7 g glucose could be produced from 1 kg rice straw with the co-production of 3.84 g FA and 6.98 g p-CA after alkaline pretreatment. These results show that the recovery of phenolic compounds from the residual stream could be a green strategy for the low-cost bioconversion of rice straw.

Influence of rice straw-derived dissolved organic matter on lactic acid fermentation by Rhizopus oryzae.

Rice straw can be used as carbon sources for lactic acid fermentation. However, only a small amount of lactic acid is produced even though Rhizopus oryzae can consume glucose in rice straw-derived hydrolysates. This study correlated the inhibitory effect of rice straw with rice straw-derived dissolved organic matter (DOM). Lactic acid fermentations with and without DOM were conducted to investigate the effect of DOM on lactic acid fermentation by R. oryzae. Fermentation using control medium with DOM showed a similar trend to fermentation with rice straw-derived hydrolysates, showing that DOM contained the major inhibitor of rice straw. DOM assay indicated that it mainly consisted of polyphenols and polysaccharides. The addition of polyphenols and polysaccharides derived from rice straw confirmed that lactic acid fermentation was promoted by polysaccharides and significantly inhibited by polyphenols. The removal of polyphenols also improved lactic acid production. However, the loss of polysaccharides during the removal of polyphenols resulted in low glucose consumption. This study is the first to investigate the effects of rice straw-derived DOM on lactic acid fermentation by R. oryzae. The results may provide a theoretical basis for identifying inhibitors and promoters associated with lactic acid fermentation and for establishing suitable pretreatment methods.

Use of magnetic powder to effectively improve the performance of sequencing batch reactors (SBRs) in municipal wastewater treatment.

This study aims to investigate the effect of adding magnetic powder in the sequencing batch reactor (SBR) on the reactor performance and microbial community. Results indicated that, the magnetic activated sludge sequencing batch reactor (MAS-SBR) had 7.76% and 4.76% higher ammonia nitrogen (NH4+-N) and chemical oxygen demand (COD) removal efficiencies than that of the conventional SBR (C-SBR). The MAS-SBR also achieved 6.86% sludge reduction compared with the C-SBR. High-throughput sequencing demonstrated that the dominant phyla of both SBRs (present as ≥1% of the sequence reads) were Protebacteria, Bacteroidetes, Chloroflexi, Saccharibacteria, Chlorobi, Firmicutes, Actinobactoria, Acidobacteria, Planctomycetes and unclassified_Bacteria. The relative abundance of Protebacteria and Bacteroidetes simultaneously declined whereas the other 8 phyla increased following the addition of magnetic powder. Adding magnetic powder in the SBR significantly affected the microbial diversity and richness of activated sludge, consequently affecting the reactor performance.

Effect of magnetic powder on membrane fouling mitigation and microbial community/composition in membrane bioreactors (MBRs) for municipal wastewater treatment.

This study aims to investigate the usefulness of magnetic powder addition in membrane bioreactors (MBRs) for membrane fouling mitigation and its effect on microbial community and composition. The comparison between the two MBRs (one with magnetic powder (MAS-MBR) and one without magnetic powder (C-MBR)) was carried out to treat synthetic municipal wastewater. Results showed that bioflocculation and adsorption of magnetic powder contributed only minimally to membrane fouling mitigation while the slower fouling rate might be ascribed to magnetic bio-effect. The macromolecules (larger than 500 kDa and 300-500 kDa) of soluble microbial product from the MAS-MBR were reduced by 24.06% and 11.11%, respectively. High-throughput sequencing demonstrated the most abundant genera of biofilm sludge indicated lower abundance in bulk sludge from the MAS-MBR compared to the C-MBR. It is possible that less membrane fouling is connected to reductions in large molecules and pioneer bacteria from bulk sludge.

Inhibitory effects of phenolic compounds of rice straw formed by saccharification during ethanol fermentation by Pichia stipitis.

In this study, it was found that the type of phenolic acids derived from rice straw was the major factor affecting ethanol fermentation by Pichia stipitis. The aim of this study was to investigate the inhibitory effect of phenolic acids on ethanol fermentation with rice straw. Different cellulases produced different ratios of free phenolic acids to soluble conjugated phenolic acids, resulting in different fermentation efficiencies. Free phenolic acids exhibited much higher inhibitory effect than conjugated phenolic acids. The flow cytometry results indicated that the damage to cell membranes was the primary mechanism of inhibition of ethanol fermentation by phenolic acids. The removal of free phenolic acids from the hydrolysates increased ethanol productivity by 2.0-fold, indicating that the free phenolic acids would be the major inhibitors formed during saccharification. The integrated process for ethanol and phenolic acids may constitute a new strategy for the production of low-cost ethanol.