Pyrolysis of Furfural Residues and Possible Utilization Pathway

The manuscript attempts to understand the evolution of NOx precursors: NH3 and HCN from Pyrolysis of furfural residue (FR). The pyrolysis process was carried out in a thermogravimetric analyzer (TGA) coupled to Fourier-transform infrared (FTIR) spectrometer. The combination revealed insightful information on the evolution of NH3 and HCN. This could help us better understand the characteristics of FR derived from furfural production especially with regard to NH3 and HCN. Nitrogen is considered a minor component in biomass wastes; in this study nitrogen content is about 0.57%. However, the pollution potential poised by low nitrogen content is huge through both direct and indirect processes. Thus, this study presents results that were found with regard to FR pyrolysis in pure nitrogen environment. At the heating rate of 40°C/min−1, the only NOx precursor detected was HCN at 713 cm−1 as per the database provided by National Institute of Standards and Technology (NIST). NH3 was not detected. The particle size of FR used ranged between 0.15–0.25 mm.

The N-Acetylglucosamine Kinase from Yarrowia lipolytica Is a Moonlighting Protein

In Yarrowia lipolytica, expression of the genes encoding the enzymes of the N-acetylglucosamine (NAGA) utilization pathway (NAG genes) becomes independent of the presence of NAGA in a Ylnag5 mutant lacking NAGA kinase. We addressed the question of whether the altered transcription was due to a lack of kinase activity or to a moonlighting role of this protein. Glucosamine-6-phosphate deaminase (Nag1) activity was measured as a reporter of NAG genes expression. The NGT1 gene encoding the NAGA transporter was deleted, creating a Ylnag5 ngt1 strain. In glucose cultures of this strain, Nag1 activity was similar to that of the Ylnag5 strain, ruling out the possibility that NAGA derived from cell wall turnover could trigger the derepression. Heterologous NAGA kinases were expressed in a Ylnag5 strain. Among them, the protein from Arabidopsis thaliana did not restore kinase activity but lowered Nag1 activity 4-fold with respect to a control. Expression in the Ylnag5 strain of YlNag5 variants F320S or D214V with low kinase activity caused a repression similar to that of the wild-type protein. Together, these results indicate that YlNag5 behaves as a moonlighting protein. An RNA-seq analysis revealed that the Ylnag5 mutation had a limited transcriptomic effect besides derepression of the NAG genes.

Improvement of putrescine production through the arginine decarboxylase pathway in Escherichia coli K-12

In the bio-based polymer industry, putrescine is in the spotlight for use as a material. We constructed strains of Escherichia coli to assess its putrescine production capabilities through the arginine decarboxylase pathway in batch fermentation. N-Acetylglutamate (ArgA) synthase is subjected to feedback inhibition by arginine. Therefore, the 19th amino acid residue, Tyr, of argA was substituted with Cys to desensitize the feedback inhibition of arginine, resulting in improved putrescine production. The inefficient initiation codon GTG of argA was substituted with the effective ATG codon, but its replacement did not affect putrescine production. The essential genes for the putrescine production pathway, speA and speB, were cloned into the same plasmid with argAATG Y19C to form an operon. These genes were introduced under different promoters; lacIp, lacIqp, lacIq1p, and T5p. Among these, the T5 promoter demonstrated the best putrescine production. In addition, disruption of the puuA gene encoding enzyme of the first step of putrescine degradation pathway increased the putrescine production. Of note, putrescine production was not affected by the disruption of patA, which encodes putrescine aminotransferase, the initial enzyme of another putrescine utilization pathway. We also report that the strain KT160, which has a genomic mutation of YifEQ100TAG, had the greatest putrescine production. At 48 h of batch fermentation, strain KT160 grown in terrific broth with 0.01 mM IPTG produced 19.8 mM of putrescine.

Valorization of Urban Street Tree Pruning Residues in Biorefineries by Steam Refining: Conversion Into Fibers, Emulsifiers, and Biogas

Street tree pruning residues are a widely available and currently undervalorized bioresource. Their utilization could help alleviate an increasing biomass shortage and offset costs of the pruning process for the municipalities. In this work, a holistic valorization pathway of pruning residues leading to fibers, oligosaccharides, biogas, and compost is presented. For this, representative mixtures of tree pruning materials from the most prevalent street tree genera (oak, linden, maple) found in Hamburg (Germany) were prepared by shredding and cleaning procedures. Collection of sample material was performed in summer and winter to account for seasonality. A steam-based fractionation was conducted using treatment severities ranging from log R0 = 2.5 to 4.0. At the highest severity, a fiber yield of around 66%, and liquor yield of 26–30% was determined. The fibers were evaluated with respect to their properties for paper product applications, with higher treatment severities leading to higher paper strengths. From the oligosaccharide-rich liquor, emulsions were created, which showed promising stability properties over 8 weeks of storage. The liquors and the rejects from the material preparation also displayed good potential for biomethane production. Overall, the differences between material collected in summer and winter were found to be small, indicating the possibility for a year-round utilization of pruning residues. For the presented utilization pathway, high severity treatments were the most promising, featuring a high liquor yield, good biomethane potential, and the highest paper strengths.

Developing an Ethanol Utilization Pathway based NADH Regeneration System in Escherichia coli

Interests remain in searching for cofactor regeneration system with higher efficiency at lower substrate cost. Glucose dehydrogenase (GDH) system has been dominant in NADH regeneration, but it only has a theoretical yield of one NADH per glucose molecule. This work sought to explore the utility of a two-step ethanol utilization pathway (EUP) in pathway-based NADH regeneration. The pathway runs from ethanol to acetaldehyde and to acetyl-CoA with each step generating one NADH, that together results in a higher theoretical yield of two NADH per ethanol molecule. In this project, anaerobic biotransformation of ketone (acetophenone or butanone) to alcohol by cpsADH from Candida parapsilosis was used as readout for evaluating relative efficacy and operating modes for EUP cofactor regeneration in Escherichia coli BL21 (DE3). Experiment tests validated that EUP was more efficient than GDH in NADH regeneration. Further, growing cell delivered higher biotransformation efficiency compared to resting cell due to the driving force generated by cell growth. Finally, preculture or cultivation in M9 + 10 g/L ethanol medium delivered higher biotransformation efficiency compared to LB medium. Overall, EUP could help regenerate NADH in support of a biocatalytic reaction, and is more efficient in cofactor regeneration than GDH.

Nicotinate degradation in a microbial eukaryote: a novel, complete pathway extant in Aspergillus nidulans

Several strikingly different aerobic and anaerobic pathways of nicotinate utilization had been described in bacteria. No similar work is extant in any eukaryote. Here we elucidate a complete eukaryotic nicotinate utilization pathway, by constructing single or multiple gene deleted strains and identifying metabolic intermediates by ultra-high performance liquid chromatography — high-resolution mass spectrometry. Enzymes catalyzing each step and all intermediate metabolites were identified. We previously established that the cognate eleven genes organized in three clusters constitute a regulon, strictly dependent on HxnR, a pathway-specific transcription factor. The first step, hydroxylation of nicotinic acid to 6-hydroxynicotinic acid is analogous to that occurring in bacterial pathways and is catalyzed by an independently evolved molybdenum-containing hydroxylase. The following enzymatic steps have no prokaryotic equivalents: 6-hydroxynicotinic acid is converted to 2,3,6-trihydroxypyridine through 2,5-dihydroxypiridine and the trihydroxylated pyridine ring is then saturated to 5,6-dihydroxypiperidine-2-one followed by the oxidation of the C6 hydroxyl group resulting in 3-hydroxypiperidine-2,6-dione. The latter two heterocyclic compounds are newly identified cellular metabolites, while 5,6-dihydroxypiperidine-2-one is a completely new chemical compound. Ring opening between C and N results in α-hydroxyglutaramate, an unprecedented compound in prokaryotic nicotinate catabolic routes. The pathway extant in A. nidulans, and in many other ascomycetes, is different from any other previously analyzed in bacteria. Our earlier phylogenetic analysis of Hxn proteins together with the complete novel biochemical pathway we now describe further illustrates the convergent evolution of catabolic pathways between fungi and bacteria.

Recent ways of management and disposal of agricultural waste - A Review

With the development of agriculture in India, the production of agricultural wastes increased rapidly. Basically these wastes are bio products and are very important for survival of animals and human beings. The occurrence of agricultural wastes is unique in the different areas. The agricultural straw and livestock excrement are considered to be potential resources. These substances are widely available on earth and can be a good source of energy or be converted into useful products. The wastes generated from crop have a good potential to convert to energy through related energy sector. The waste produce from animal or from crop residue is called biomass which has an interdependent relationship with ecosystem from production to disposal and has physicochemical properties. The recycling and utilization of agricultural wastes are considered to be the important step in environmental protection, energy structure and agricultural development. The present review deals with the research work carried out in the conversion of biomass and agricultural waste and to illuminate the potential environmental risk, recycling and utilization pathway, influencing factors and policy suggestions in the recycling and utilization progress of agricultural wastes. An attempt is carried out to increase the economic value of agricultural waste into useful product. The survey provided the development mode of industrialization and scale of agricultural waste recycling. The recycling and utilization pathway of agricultural wastes were also analysed. The crucial suggestions may be proposed, such as cultivating new industry, building economy incentive standard, improving laws and regulations, and creating rural market strengthening medium and long-term plans of agricultural waste recycling. The resource consumption, ecological crisis and other issues caused by agricultural wastes were evaluated.

ATP-dependent protease ClpP and its subunits ClpA, ClpB, and ClpX involved in the field bismerthiazol-resistance in Xanthomonas oryzae pv. oryzae

Resistance of Xanthomonas oryzae pv. oryzae (Xoo), which causes rice bacterial leaf blight, to bismerthiazol has been detected in China since the 1990s. Interestingly, the strains resistant to bismerthiazol on rice plants were more sensitive to bismerthiazol than wild-type (WT) strains in vitro. Here, qPCR was applied to detect the fold expression of ATP-dependent proteases, ClpP and its subunits, which cope with stresses including bactericides in bismerthiazol-resistant strains and their parental susceptible WT strain (ZJ173). Results showed that the expression of ClpP and its subunits were all higher in bismerthiazol-resistant strains than that in ZJ173. They were up-regulated during early growth phase and down-regulated during middle growth phase in ZJ173 treated with bismerthiazol but did not change in the resistant strains. ClpP and its subunits were overexpressed in Xoo in this study, the higher expression of these genes increased sensitivity in vitro and enhanced resistance in vivo to bismerthiazol. Inhibition of bismerthiazol to exopolysaccharide production, biofilm, and motility was also reduced in ClpP and its subunit’s overexpression mutants of Xoo. The deletion mutants of ClpP and its subunits in ZJ173 abated pathogenicity, biofilm production, swimming ability, exopolysaccharide production, and growth in low-nutrient environments. Moreover, ClpP and its subunits may act downstream of the histidine utilization pathway, which could be inhibited by bismerthiazol in Xoo. Taken together, our results indicated that ClpP and its subunits of Xoo influenced the resistance to bismerthiazol.