Reactivity improvement of cellulolytic enzyme lignin via mild hydrothermal modification

Isolated by the cellulolytic enzyme lignin (CEL) process, water-alcohol (1:1, v/v) was introduced as co-solvent in the process of the hydrothermal treatment. The modification parameters such as reaction temperature and time, solid-to-liquid ratio, and catalysts (NaOH and NaOAlO₂) have been investigated in terms of the specific lignin properties, such as the phenolic hydroxyl content (OH_phen), DPPH free radical scavenging rate, and formaldehyde value. The CELs were also characterized by GPC, FT-IR and ¹ H NMR spectroscopy, and Py-GC/MS. The key data are under optimal lignin modification conditions (solid-to-liquid ratio of 1:10 (w/v) and a temperature of 250 °C for 60 min) are: OH_phen content: 2.50 mmol/g; half maximal inhibitory concentration (IC₅₀) towards DPPH free radicals: 88.2 mg/L; formaldehyde value: 446.9 g/kg). Both base catalysts decrease the residue rate, but phenol reactivities of the products were also detracted. Py-GC/MS results revealed that modified lignin had a higher phenolic composition than the CEL did, especially the modified lignin without catalyst (ML), which represented 74.51% phenolic content.     

Optimized pulse-feeding fed-batch fermentation for enhanced lignin to polyhydroxyalkanoate transformation

With an anticipated growth of Bio-Circular-Green economy, the amount of lignin generated as by-product from biorefineries is increasing. Hence, lignin valorising strategies become a very interesting option to improve economic viability of the biorefineries. This study revealed the development of bioprocesses for upgrading lignin into bioplastic. Specifically, a novel strain of Pseudomonas fulva has been applied for microbial bioconversion of organosolv lignin to fermentative polyhydroxyalkanoate (PHA) production. Fed-batch fermentation of lignin-to-PHA with pulse-feeding approach was optimized using Design of Experiment. Effects of C:N ratio and feeding profiles on PHA accumulation in P. fulva were investigated to determine optimal operation. Under optimized fed-batch, the PHA concentration of 195.2 ± 6.6 mg/L could be reached and the PHA content was more than 2 folds enhancement compared to batch process. Type of PHA produced was also characterized for chemical composition via GC–MS analysis. The established lignin to PHA conversion could provide platform for developing integrated lignin bioprocessing to promote cost-effective biorefineries.     

Protocol for assessing soybean antixenosis to Heliothis virescens

Larvae of Heliothis virescens (Fabricius) (Lepidoptera: Noctuidae) often infest soybean crops, Glycine max (L.) (Fabaceae), causing significant yield losses in important soybean-producing regions. The use of soybean varieties resistant to lepidopteran larvae is a major approach in soybean integrated pest management. However, standardization and optimization of bioassays that are used to screen genotypes for insect resistance are essential for high-throughput phenotyping. Methodologies for screening were assessed to determine the most effective method for discriminating levels of antixenosis to H. virescens in soybean plants. Feeding and oviposition preference assays were performed to determine optimal densities of larvae and adults, and optimal plant structures and growth stages for conducting assays. In addition, trichome densities, and fiber and lignin contents were quantified in plant structures of soybean cultivars differing in resistance. Resistance levels of cultivars were best differentiated using nine neonate larvae and two 6-day-old larvae, and by using young leaves of plants at the vegetative stage. This was likely due to the more pronounced differences in lignin and fiber contents in young leaves of vegetative-stage plants. Density of adult pairs, plant structure, and growth stage did not affect ability to distinguish differences in oviposition preference by H. virescens. Higher numbers of eggs were found on the leaves, which were the plant structures that exhibited the lowest trichome densities. The protocol developed in this work will benefit future evaluations of soybean genotypes for antixenosis against H. virescens.     

Regulatory role of cinnamyl alcohol dehydrogenase in the formation of guaiacyl and syringyl lignins

The substrate specificities of cinnamyl alcohol dehydrogenase (CAD) of angiosperms and gymnosperms were examined using coniferaldehyde and sinapaldehyd     

Soil organic carbon content and composition of 130-year crop, pasture and forest land-use managements

Conversion of former agricultural land to grassland and forest ecosystems is a suggested option for mitigation of increased atmospheric CO₂. A Sharpsburg prairie loess soil (fine, smectitic, mesic Typic Argiudoll) provided treatments to study the impact of long‐term land use on soil organic carbon (SOC) content and composition for a 130‐year‐old cropped, pasture and forest comparison. The forest and pasture land use significantly retained more SOC, 46% and 25%, respectively, compared with cropped land use, and forest land use increased soil C content by 29% compared with the pasture. Organic C retained in the soils was a function of the soil N content (r=0.98, P<0.001) and the soil carbohydrate (CH) concentration (r=0.96, P<0.001). Statistical analyses found that soil aggregation processes increased as organic C content increased in the forest and pasture soils, but not in the cropped soil. SOC was composed of similar percentages of CHs (49%, 42% and 51%), amino acids (22%, 15% and 18%), lipids (2.3%, 2.3% and 2.9%) and unidentified C (21%, 29% and 27%), but differed for phenolic acids (PAs) (5.7%, 11.6% and 1.0%) for the pasture, forest and cropped soils, respectively. The results suggested that the majority of the surface soil C sequestered in the long‐term pasture and forest soils was identified as C of plant origin through the use of CH and PA biomarkers, although the increase in amino sugar concentration of microbial origin indicates a greater increase in microbial inputs in the three subsoils. The practice of permanent pastures and afforestation of agricultural land showed long‐term potential for potential mitigation of atmospheric CO₂.     

Variable silicon accumulation in plants affects terrestrial carbon cycling by controlling lignin synthesis

Current climate and land‐use changes affect regional and global cycles of silicon (Si), with yet uncertain consequences for ecosystems. The key role of Si in marine ecology by controlling algae growth is well recognized but research on terrestrial ecosystems neglected Si since not considered an essential plant nutrient. However, grasses and various other plants accumulate large amounts of Si, and recently it has been hypothesized that incorporation of Si as a structural plant component may substitute for the energetically more expensive biosynthesis of lignin. Herein, we provide evidence supporting this hypothesis. We demonstrate that in straw of rice (Oryza sativa) deriving from a large geographic gradient across South‐East Asia, the Si concentrations (ranging from 1.6% to 10.7%) are negatively related to the concentrations of carbon (31.3% to 42.5%) and lignin‐derived phenols (32 to 102 mg/g carbon). Less lignin may explain results of previous studies that Si‐rich straw decomposes faster. Hence, Si seems a significant but hardly recognized factor in organic carbon cycling through grasslands and other ecosystems dominated by Si‐accumulating plants.     

A New Lignan (Polonilignan) and Inhibitors of Nitric Oxide Production from Penicillium polonicum,an Endophytic Fungi of Piper nigrum

Inhibiting nitric oxide (NO) or its production is found to be of therapeutic benefit. To discover natural small molecule inhibitors of NO production, a bioassay- and LC/MS-guided chemical investigation was done on the metabolites of endophytic fungus isolated from fresh Piper nigrum fruits. The isolated pure strain was identified as Penicillium polonicum by 16S rDNA sequence comparison. The culture broth extract of P. polonicum (EEPP) exhibited a significant reduction of NO production (Griess method) in LPS-stimulated RAW 264.7 cells (P<0.0001). To understand the chemical constituents of bioactive EEPP, column chromatography and p-TLC studies were carried out, which yielded eight pure compounds. They were characterised as botryosphaeridione ( 1 ), 3-(3,5-di-tert-butyl-4-hydroxy)phenylpropionic acid ( 2 ), variabilone ( 3 ), 2,4-di-tert-butylphenol ( 4 ), indole-3-carboxylic acid ( 5 ), tyrosol ( 6 ), ethyl ferulate ( 7 ) and a new lignan ( 8 ) based on the spectral analysis. The structure elucidation of the new lignan, named polonilignan ( 8 ), was based on HR-MS, ¹H- & ¹³C-NMR, H−H COSY, HSQC and HMBC spectra. Compounds 2 , 4 , 5 and 6 showed a significant decrease (P<0.0001) in the production of NO in LPS-induced RAW 264.7 cells. Tyrosol ( 6 ) and indole-3-carboxylic acid ( 5 ) controlled nitrite release with IC₅₀ values of 22.84 and 55.01 μM, respectively. This is the first report of (i) P. polonicum as an endophytic fungus of pepper fruits, (ii) isolation of compounds 1 – 8 except 6 from P. polonicum culture broth extract and (iii) NO inhibition effect of 2 , 4 , 5 and 6 .