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The use of metal nanoparticles (NPs) to enhance hydrogen production in dark fermentation (DF) has become a pioneering field of interest. In particular, iron-based nanoparticles (FeNPs) play a pivotal role in enhancing the activity of metalloenzymes and optimizing feedstock utilization, resulting in improved hydrogen production. This study investigated the effect of FeNPs (magnetite) supplementation at three different concentrations of 50, 100, and 200 ppm in a continuous dark fermenter for the production of hydrogen from rice straw acid hydrolysate. The highest hydrogen production rate of 2.6 ± 0.3 NL H2/L-d was achieved with the addition of 100 ppm of nanoparticles, representing a 53% increase compared to the condition without FeNPs addition. This improvement was driven by a microbial community in which Clostridium was the major dominant genus. In addition, increasing the nanoparticle concentration to 100 ppm resulted in an increase in butyrate concentration to 2.0 ± 0.1 g/L, which is 43% higher than the butyrate concentration without FeNPs. However, when the NP concentration was increased to 200 ppm, the hydrogen production rate decreased to 1.6 ± 0.2 NL H2/L-d. This study can serve as a guideline for future research aimed at evaluating the effects of FeNPs in continuous dark fermentation systems. This work highlights the potential benefits and challenges associated with the use of FeNPs, paving the way for future studies to optimize their application and improve the efficiency of dark fermentation processes.

[This corrects the article DOI: 10.3389/fbioe.2021.635509.].

The production of microbial lipid using lignocellulosic agroforestry residues has attracted much attention. But, various inhibitors such as phenols and furans, which are produced during lignocellulosic hydrolysate preparation, are harmful to microbial lipid accumulation. Herein, we developed a novel detoxification strategy of rice straw hydrolysate using immobilized laccase on magnetic Fe3O4 nanoparticles for improving lipid production of Rhodotorula glutinis. Compared with free laccase, the immobilized laccase on magnetic nanoparticles showed better stability, which still retained 76% of original activity at 70 °C and 56% at pH 2 for 6 h. This immobilized laccase was reused to remove inhibitors in acid-pretreated rice straw hydrolysate through recycling with external magnetic field. The results showed that most of phenols, parts of furans, and formic acids could be removed by immobilized laccase after the first batch. Notably, the immobilized laccase exhibited good reusability in repeated batch detoxification. 78.2% phenols, 43.8% furfural, 30.4% HMF, and 16.5% formic acid in the hydrolysate were removed after the fourth batch. Furthermore, these detoxified rice straw hydrolysates, as substrates, were applied to the lipid production of Rhodotorula glutinis. The lipid yield in detoxified hydrolysate was significantly higher than that in undetoxified hydrolysate. These findings suggest that the immobilized laccase on magnetic nanoparticles has a potential to detoxify lignocellusic hydrolysate for improving microbial lipid production.

Rice straw hydrolysate produced in rice straw pretreatment, comprising a lot of fermentable sugars, is generally released into the environment. This not only causes environment pollution but also wastes fermentable sugars from rice straw. To alleviate environment impact, maximize ethanol production from rice straw, and reduce the cost of ethanol production, rice straw hydrolysate and NaOH-pretreated rice straw were converted to ethanol using ethanol-type fermentation and simultaneous saccharification and fermentation (SSF) by sludge. Meanwhile, microbial community in sludge was analyzed to find the relationship between ethanol production and microbial community succession during ethanol-type fermentation and SSF. Under the optimal condition of the COD, pH and oxidation–reduction potential (ORP) value of rice straw hydrolysate with 6280.56 mg/L, 6.7, and − 42 mV, ethanol-type fermentation with the sludge of 15 g obtained the highest ethanol concentration (8.34 g/L) and the highest COD removal rate (54.83%). For SSF, the maximum ethanol concentration (3.75 g/L) produced by pretreated rice straw and sludge from ethanol-type fermentation with the sludge of 15 g was higher than that (2.61 g/L) generated by pretreated rice straw and sludge from ethanol-type fermentation with the sludge of 22.5 g. This indicated that sludge from ethanol-type fermentation with the sludge of 15 g more efficiently converted rice straw to ethanol than sludge from ethanol-type fermentation with the sludge of 22.5 g. Microbial community analysis suggested that ethanol production had a negative correlation with the relative abundance changes of Bacteroidetes, when the relative abundance of Firmicutes constantly rose in ethanol-type fermentation and SSF. This study provides a scientific basis for maximizing ethanol production from rice straw by microbial regulation in sludge, which could further reduce the cost of ethanol production.

Rice straw and sugar cane industrial waste are a plentiful source of lignocellulosic biomass with a high polysaccharide content, that is hydrolyzed into sugar for microbial growth and their metabolites. 3-Hydroxypropionic acid (3-HP) is a promising chemical building block that can be produced from renewable resources. The malonyl-CoA pathway is one of the biosynthetic routes for 3-HP production by expressing the malonyl-CoA reductase gene (mcr). However, the problem of the activity imbalance between the C and N-terminal causes a low conversion rate of malonyl-CoA to 3-hydroxypropionic acid. This study aimed to balance the bi-functional MCR enzyme by dissecting MCR into two fragments and enhancing the supply of intermediates to increase the production of 3-HP. The recombinant strain harboring the dissected mcr gene showed a 21-fold increase in 3-HP titer compared to the strain carrying the full-length mcr gene. The addition of cerulenin and acetate to the fermented medium enhanced 3-HP yield by 8 times, in which recombinant yeast produced 3-HP up to 10 g/L (0.201 gproduct/gsubstrate). The results of using rice straw hydrolysate as a carbon source indicated that Saccharomyces cerevisiae S2 produced 3-HP of 4.02 g/L, which was 0.074 gproduct/gglucose in the diluted hydrolysate. These findings provide an alternative and sustainable strategy for utilizing lignocellulosic biomass for future 3-HP production at an industrial scale.

The non-proteinogenic amino acid 5-amino valeric acid (5-AVA) and the diamine putrescine are potential building blocks in the bio-polyamide industry. The production of 5-AVA and putrescine using engineered Corynebacterium glutamicum by the co-consumption of biomass-derived sugars is an attractive strategy and an alternative to their petrochemical synthesis. In our previous work, 5-AVA production from pure xylose by C. glutamicum was shown by heterologously expressing xylA from Xanthomonas campestris and xylB from C. glutamicum . Apart from this AVA Xyl culture, the heterologous expression of xylA Xc and xylB Cg was also carried out in a putrescine producing C. glutamicum to engineer a PUT Xyl strain. Even though, the pure glucose (40 g L –1 ) gave the maximum product yield by both the strains, the utilization of varying combinations of pure xylose and glucose by AVA Xyl and PUT Xyl in CGXII synthetic medium was initially validated. A blend of 25 g L –1 of glucose and 15 g L –1 of xylose in CGXII medium yielded 109 ± 2 mg L –1 putrescine and 874 ± 1 mg L –1 5-AVA after 72 h of fermentation. Subsequently, to demonstrate the utilization of biomass-derived sugars, the alkali (NaOH) pretreated-enzyme hydrolyzed rice straw containing a mixture of glucose (23.7 g L –1 ) and xylose (13.6 g L –1 ) was fermented by PUT Xyl and AVA Xyl to yield 91 ± 3 mg L –1 putrescine and 260 ± 2 mg L –1 5-AVA, respectively, after 72 h of fermentation. To the best of our knowledge, this is the first proof of concept report on the production of 5-AVA and putrescine using rice straw hydrolysate (RSH) as the raw material.

BackgroundBioethanol obtained by fermenting cellulosic fraction of biomass holds promise for blending in petroleum. Cellulose hydrolysis yields glucose while hemicellulose hydrolysis predominantly yields xylose. Economic feasibility of bioethanol depends on complete utilization of biomass carbohydrates and an efficient co-fermenting organism is a prerequisite. While hexose fermentation capability of Saccharomyces cerevisiae is a boon, however, its inability to ferment pentose is a setback.ResultsTwo xylose fermenting Kodamaea ohmeri strains were isolated from Lagenaria siceraria flowers through enrichment on xylose. They showed 61% glucose fermentation efficiency in fortified medium. Medium engineering with 0.1% yeast extract and peptone, stimulated co-fermentation potential of both strains yielding maximum ethanol 0.25 g g−1 on mixed sugars with ~ 50% fermentation efficiency. Strains were tolerant to inhibitors like 5-hydroxymethyl furfural, furfural and acetic acid. Both K. ohmeri strains grew well on biologically pretreated rice straw hydrolysates and produced ethanol.ConclusionsThis is the first report of native Kodamaea sp. exhibiting notable mixed substrate utilization and ethanol fermentation. K. ohmeri strains showed relevant traits like utilizing and co-fermenting mixed sugars, exhibiting excellent growth, inhibitor tolerance, and ethanol production on rice straw hydrolysates.

Bacterial synthesis of 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV) copolymer [P(3HB- co -3HV)] using the hydrolysate of rice straw waste as a carbon source was affected by the composition of the hydrolysate, which depends highly on the rice straw pretreatment condition. Acid digestion with 2 % sulfuric acid generated larger production of P(3HB- co -3HV) than 6 % sulfuric acid, but 3HV concentration in the copolymer produced with 2 % acid hydrolysate was only 8.8 % compared to 18.1 % with 6 % acid hydrolysate. To obtain a higher 3HV mole fraction for enhanced flexibility of the copolymer, an additional heating was conducted with the 2 % acid hydrolysate after removal of residual rice straw. As the additional heating time increased a higher concentration of levulinic acid was generated, and consequently, the mole fraction of 3HV in P(3HB- co -3HV) increased. Among the conditions tested (i.e., 20-, 40-, 60-min), 60-min additional heating following 2 % sulfuric acid digestion achieved the highest 3HV mole fraction of 22.9 %. However, a longer heating time decreased the P(3HB- co -3HV) productivity, probably due to the increased intermediates concentrations acting as inhibitors in the hydrolysates. Therefore, the use of additional heating needs to consider both the increase in the 3HV mole fraction and the decrease in the P(3HB- co -3HV) productivity.

Doc number: 23 Abstract Background: The use of lignocellulosic constituents in biotechnological processes requires a selective separation of the main fractions (cellulose, hemicellulose and lignin). During diluted acid hydrolysis for hemicellulose extraction, several toxic compounds are formed by the degradation of sugars and lignin, which have ability to inhibit microbial metabolism. Thus, the use of a detoxification step represents an important aspect to be considered for the improvement of fermentation processes from hydrolysates. In this paper, we evaluated the application of Advanced Oxidative Processes (AOPs) for the detoxification of rice straw hemicellulosic hydrolysate with the goal of improving ethanol bioproduction by Pichia stipitis yeast. Aiming to reduce the toxicity of the hemicellulosic hydrolysate, different treatment conditions were analyzed. The treatments were carried out according to a Taguchi L16 orthogonal array to evaluate the influence of Fe+2 , H2 O2 , UV, O3 and pH on the concentration of aromatic compounds and the fermentative process. Results: The results showed that the AOPs were able to remove aromatic compounds (furan and phenolic compounds derived from lignin) without affecting the sugar concentration in the hydrolysate. Ozonation in alkaline medium (pH 8) in the presence of H2 O2 (treatment A3) or UV radiation (treatment A5) were the most effective for hydrolysate detoxification and had a positive effect on increasing the yeast fermentability of rice straw hemicellulose hydrolysate. Under these conditions, the higher removal of total phenols (above 40%), low molecular weight phenolic compounds (above 95%) and furans (above 52%) were observed. In addition, the ethanol volumetric productivity by P. stipitis was increased in approximately twice in relation the untreated hydrolysate. Conclusion: These results demonstrate that AOPs are a promising methods to reduce toxicity and improve the fermentability of lignocellulosic hydrolysates.

A detoxification method using activated charcoal with concentrated rice straw hemicellulosic hydrolysate improved the conversion of xylose to xylitol by the yeast Candida guilliermondii by 22%. This was achieved when the hydrolysate:charcoal ratio was 40 g g^sup -1^, resulting in removal of 27% of phenolic compounds. Under this condition, the xylitol yield factor (0.72 g g^sup -1^) and volumetric productivity (0.61 g l^sup -1^ h^sup -1^) were close to those attained in a semi-defined medium simulating hydrolysate sugars.[PUBLICATION ABSTRACT]