Exploring the emulsification potential of chitosan modified with phenolic acids: Emulsifying properties, functional activities, and application in curcumin encapsulation.

This study successfully grafted caffeic acid and 3,4-dihydroxybenzoic acid into chitosan through a coupling reaction, yielding grafting ratio of 8.93 % for caffeic acid grafted chitosan (CA-GC) and 9.15 % for 3,4-dihydroxybenzoic acid grafted chitosan (DHB-GC) at an optimal concentration of 4 mmol phenolic acids. The characterization of modified chitosans through ultraviolet visible spectrometer (UV-vis), Fourier transform infrared spectrometer (FTIR), proton nuclear magnetic resonance (1H NMR), and x-ray photoelectron spectrometer (XPS) confirmed the successful grafting of phenolic acids. In the subsequent step of emulsion preparation, confocal laser scanning microscope images confirmed the formation of O/W (oil-in-water) emulsions. The phenolic acid-grafted chitosans exhibited better emulsification properties compared to native chitosan, such as reduced droplet size, more uniform emulsion droplet distribution, increased ζ-potential, and enhanced emulsifying activity and stability. Moreover, the modified chitosans demonstrated increased antioxidant activities (evidenced by DPPH and ß-carotene assays) and displayed greater antimicrobial effects against E. coli and S. aureus. Its efficacy in curcumin encapsulation was also notable, with improved encapsulation efficiency, sustained release rates, and enhanced storage and photostability. These findings hint at the potential of modified chitosans as an effective emulsifier.

2-Furoic acid associated with the infection of nematodes by Dactylellina haptotyla and its biocontrol potential on plant root-knot nematodes.

Dactylellina haptotyla is a typical nematode-trapping fungus that has garnered the attention of many scholars for its highly effective lethal potential for nematodes. Secondary metabolites play an important role in D. haptotyla-nematode interactions, but which metabolites perform which function remains unclear. We report the metabolic functions based on high-quality, chromosome-level genome assembly of wild D. haptotyla YMF1.03409. The results indicate that a large variety of secondary metabolites and their biosynthetic genes were significantly upregulated during the nematode-trapping stage. In parallel, we identified that 2-furoic acid was specifically produced during nematode trapping by D. haptotyla YMF1.03409 and isolated it from fermentation production. 2-Furoic acid demonstrated strong nematicidal activity with an LD50 value of 55.05 µg/mL against Meloidogyne incognita at 48 h. Furthermore, the pot experiment showed that the number of galls of tomato root was significantly reduced in the experimental group treated with 2-furoic acid. The considerable increase in the 2-furoic acid content during the infection process and its virulent nematicidal activity revealed an essential synergistic effect during the process of nematode-trapping fungal infection. IMPORTANCE Dactylellina haptotyla have significant application potential in nematode biocontrol. In this study, we determined the chromosome-level genome sequence of D. haptotyla YMF1.03409 by long-read sequencing technology. Comparative genomic analysis identified a series of pathogenesis-related genes and revealed significant gene family contraction events during the evolution of D. haptotyla YMF1.03409. Combining transcriptomic and metabolomic data as well as in vitro activity test results, a compound with important application potential in nematode biocontrol, 2-furoic acid, was identified. Our result expanded the genetic resource of D. haptotyla and identified a previously unreported nematicidal small molecule, which provides new options for the development of plant biocontrol agents.

Pectin grafted with resorcinol and 4-hexylresorcinol: Preparation, characterization and application in meat preservation.

To augment the functional attributes of pectin and expand its prospective utilization in food preservation, this research explored the enzymatic grafting of resorcinol and 4-hexylresorcinol onto pectin. Structural analysis verified the successful grafting of both resorcinol and 4-hexylresorcinol to pectin via esterification, with the 1-OH of resorcinol and 4-hexylresorcinol and the carboxyl group of pectin functioning as grafting sites. The grafting ratios of resorcinol-modified pectin (Re-Pe) and 4-hexylresorcinol-modified pectin (He-Pe) were 17.84 % and 10.98 %, respectively. This grafting modification notably enhanced the antioxidative and antibacterial properties of pectin. Specifically, DPPH clearance and the inhibition ratio in the ß-carotene bleaching assay increased from 11.38 % and 20.13 % (native pectin, Na-Pe) to 41.15 % and 36.67 % (Re-Pe), and 74.72 % and 53.40 % (He-Pe). Moreover, the inhibition zone diameter against Escherichia coli and Staphylococcus aureus rose from 10.12 and 10.08 mm (Na-Pe) to 12.36 and 11.52 mm (Re-Pe), and 16.78 and 14.87 mm (He-Pe). Additionally, the application of native and modified pectin coatings effectively impeded pork spoilage, with the modified pectins demonstrating a more potent effect. Among the two modified pectins, He-Pe exhibited the most significant enhancement in pork shelf life.

Identification and Mechanism of Action of the Global Secondary Metabolism Regulator SaraC in Stereum hirsutum.

DNA methylation is an important factor in the regulation of gene expression. In analyzing genomic data of Stereum hirsutum FP-91666, we found a hypothetical bifunctional transcription regulator/O6Meguanine-DNA methyltransferase (named SaraC), which is widely present in both bacteria and fungi, and confirmed that its function in bacteria is mainly for DNA reparation. In this paper, we confirmed that SaraC has the function of DNA binding and demethylation through surface plasma resonance and reaction experiments in vitro. Then, we achieved the overexpression of SaraC (OES) in S. hirsutum, sequenced the methylation and transcription levels of the whole-genome, and further conducted untargeted metabolomics analyses of the OES transformants and the wild type (WT). The results confirmed that the overall-methylation levels of the transformants were significantly downregulated, and various genes related to secondary metabolism were upregulated. Through comparative untargeted metabolomic analyses, it showed that OES SA6 transformant produced a greater number of hybrid polyketides, and we identified 2 novel hybrid polyketides from the fermentation products of SA6. Our results show that overexpression SaraC can effectively stimulate the expression of secondary-metabolism-related genes, which could be a broad-spectrum tool for discovery of metabolites due to its cross-species conservation. IMPORTANCE Fungi are one of the important sources of active compounds. However, in fungi, most of the secondary metabolic biosynthetic gene clusters are weakly expressed or silenced under conventional culture conditions. How to efficiently excavate potential new compounds contained in fungi is becoming a research hot spot in the world. In this study, we found a DNA demethylation protein (SaraC) and confirmed that it is a global secondary metabolism regulator in Stereum hirsutum FP-91666. In the past, SaraC-like proteins were mainly regarded as DNA repair proteins, but our findings proved that it will be a powerful tool for mining secondary metabolites for overexpression of SaraC, which can effectively stimulate the expression of genes related to secondary metabolism.

Comparison of intrarenal pressure between convention and vacuum-assisted ureteral access sheath using an ex vivo porcine kidney model.

OBJECTIVE: This study aimed to prove the vacuum-assisted ureteral access sheath (vaUAS) is more effective in maintaining a lower IRP than conventional ureteral access sheath (cUAS). MATERIALS: The model consisted of 12 freshly harvested adult porcine kidneys. METHODS: Either a 12/14F cUAS or vaUAS was alternately inserted into the ureter to one cm below the renal pelvis. Upper, middle, and lower calyces were punctured, and 6F pressure monitor catheters were introduced. IRP with cUAS was monitored using various irrigation rates. IRP with vaUAS was monitored with the same irrigation rates; various aspiration pressures; and vent fully closed, 50% closed, and fully open. RESULTS: cUAS with irrigation rate of 50 cc/min resulted in IRP < 30 mmHg. 50 to 100 cc/min should be used with caution. When irrigation rate exceeded 100 cc/min, IRP rose to ≥ 30 mmHg in most instances. With vent closed, vaUAS with vacuum pressure ≥ 150 mmHg and irrigation rate of 50 cc, 100 cc, and 150 cc/min generally resulted in IRPs < 5 mmHg. With vent half closed, vaUAS with vacuum pressure ≥ 300 mmHg and irrigation rate of ≤ 100 cc/min avoided IRP > 30 mmHg. vaUAS with vent open showed limited advantages over cUAS. CONCLUSION: vaUAS maintains lower IRP than cUAS under same parameters. Both vaUAS and cUAS can be used when irrigation is ≤ 50 cc/min vaUAS showed clear advantages over cUAS in maintaining lower pressure when irrigation rate is ≥ 100 cc/min.

Stereumamides E-H, four new minor quaternary ammonium hybrids from Stereum hirsutum.

In our preview research, four novel quaternary ammonium hybrids were isolated from the Stereum hirsutum FP-91666. To further discover this type of compounds, S. hirsutum was fermented in 30 L YMG broth, and eight hybrid compounds including four new quaternary ammoniums were obtained, which are sesquiterpenes combined with α-amino acids. Their structures were elucidated by extensive spectroscopic analyses, including 1D- and 2D-NMR, and HR-MS experiments.

Recent Advances in the Synthetic Biology of Natural Drugs.

Natural drugs have been transformed and optimized during the long process of evolution. These compounds play a very important role in the protection of human health and treatment of human diseases. Sustainable approaches to the generation of raw materials for pharmaceutical products have been extensively investigated in drug research and development because chemical synthesis is costly and generates pollution. The present review provides an overview of the recent advances in the synthetic biology of natural drugs. Particular attention is paid to the investigations of drugs that may be mass-produced by the pharmaceutical industry after optimization of the corresponding synthetic systems. The present review describes the reconstruction and optimization of biosynthetic pathways for nine drugs, including seven drugs from plant sources and two drugs from microbial sources, suggesting a new strategy for the large-scale preparation of some rare natural plant metabolites and highly bioactive microbial compounds. Some of the suggested synthetic methods remain in a preliminary exploration stage; however, a number of these methods demonstrated considerable application potential. The authors also discuss the advantages and disadvantages of the application of synthetic biology and various expression systems for heterologous expression of natural drugs. Thus, the present review provides a useful perspective for researchers attempting to use synthetic biology to produce natural drugs.

Sesquiterpenoids and their quaternary ammonium hybrids from the mycelium of mushroom Stereum hirsutum by medium optimization.

The fungal genus Stereum (Stereaceae) produces a broad variety of specialised metabolites, including a wide range of terpenes. This probably relates to the presence of an extensive biosynthetic machinery for this group of compounds: genomic analysis of Stereum hirsutum has identified 16 terpene synthase gene clusters, 6 polyketide synthase gene clusters, and 1 polyketide synthase non-ribosomal polypeptide heterodimer gene cluster in S. hirsutum FP-91666. In the present study, the One Strain Many Compounds (OSMAC) approach was employed to discover undescribed metabolites from this strain. Fermentation was carried out in five media and the products of the strain cultivated on different media were analyzed by LC-MS. From cultures grow in WGB medium (30.0 g wheat bran, 20.0 g glucose, 1.5 g KH2PO4, and 1.5 g MgSO4), four previously undescribed metabolites, a sesquiterpene sterostrein X and three mixed terpenes (stereumamides I-K) were isolated, together with seven known compounds (drimene-2,11-diol, stereumamide E, stereumamide D, stereumamide B, stereumamide A, stereumamide C, and sterostrein Q). The drimane-type sesquiterpene drimene-2,11-diol was found in S. hirsutum FP-91666 for the first time. All structures were elucidated by spectroscopic data analysis. The absolute configurations of stereumamides I, J and K were assigned by comparing their experimental and calculated electronic circular dichroism (ECD) spectra. An anti-Mycobacterium tuberculosis experiment showed that stereumamides I-K and sterostrein Q had weak antibacterial activity against this pathogen.

Vib-PT, an Aromatic Prenyltransferase Involved in the Biosynthesis of Vibralactone from Stereum vibrans.

Vibralactone, a hybrid compound derived from phenols and a prenyl group, is a strong pancreatic lipase inhibitor with a rare fused bicyclic ß-lactone skeleton. Recently, a researcher reported a vibralactone derivative (compound C1) that caused inhibition of pancreatic lipase with a half-maximal inhibitory concentration of 14 nM determined by structure-based optimization, suggesting a potential candidate as a new antiobesity treatment. In the present study, we sought to identify the main gene encoding prenyltransferase in Stereum vibrans, which is responsible for the prenylation of phenol leading to vibralactone synthesis. Two RNA silencing transformants of the identified gene (vib-PT) were obtained through Agrobacterium tumefaciens-mediated transformation. Compared to wild-type strains, the transformants showed a decrease in vib-PT expression ranging from 11.0 to 56.0% at 5, 10, and 15 days in reverse transcription-quantitative PCR analysis, along with a reduction in primary vibralactone production of 37 to 64% at 15 and 21 days, respectively, as determined using ultra-high-performance liquid chromatography-mass spectrometry analysis. A soluble and enzymatically active fusion Vib-PT protein was obtained by expressing vib-PT in Escherichia coli, and the enzyme's optimal reaction conditions and catalytic efficiency (Km /kcat) were determined. In vitro experiments established that Vib-PT catalyzed the C-prenylation at C-3 of 4-hydroxy-benzaldehyde and the O-prenylation at the 4-hydroxy of 4-hydroxy-benzenemethanol in the presence of dimethylallyl diphosphate. Moreover, Vib-PT shows promiscuity toward aromatic compounds and prenyl donors.IMPORTANCE Vibralactone is a lead compound with a novel skeleton structure that shows strong inhibitory activity against pancreatic lipase. Vibralactone is not encoded by the genome directly but rather is synthesized from phenol, followed by prenylation and other enzyme reactions. Here, we used an RNA silencing approach to identify and characterize a prenyltransferase in a basidiomycete species that is responsible for the synthesis of vibralactone. The identified gene, vib-PT, was expressed in Escherichia coli to obtain a soluble and enzymatically active fusion Vib-PT protein. In vitro characterization of the enzyme demonstrated the catalytic mechanism of prenylation and broad substrate range for different aromatic acceptors and prenyl donors. These characteristics highlight the possibility of Vib-PT to generate prenylated derivatives of aromatics and other compounds as improved bioactive agents or potential prodrugs.

Surface convexity of anammox granular sludge: Digital characterization, state indication and formation mechanism.

Anammox granular sludge system is a promising biotechnology for nitrogen removal from wastewaters. The anammox granules possess the distinctive morphological features which can be developed as visible indicators for anammox process monitoring. In this study, the surface convexity of anammox granular sludge (AnGS) was systematically investigated for the first time. The AnGS was withdrawn from four long-term operated anammox bioreactors at different nitrogen loading rates. Firstly, the spherical convexity and gap concavity with a diameter of 50-100 µm were observed to be distributed on the granules surface under the microscopic observation. Then, the surface convexity was determined by image processing technology and the statistical analysis showed that the convexity had a significant difference (p = 0.003) among bioreactors and the average surface convexity decreased from 0.937 ±â€¯0.030 to 0.899 ±â€¯0.034, and then rose to 0.914 ±â€¯0.035 which had a significant correlation with the volumetric gas production rate of bioreactor (r = -0.873, p < 0.05). An optical method was further developed to fast characterize the surface convexity using relative lightness (LSCE⁎/LSCI⁎) as the index. At last, the composition and structure of AnGS were investigated to deduce the formation mechanism of surface convexity. The formation could be attributed to the outward growth of zoogloea led to the surface protrusion (convexity); the periodical extrusion of microbubbles caused the striping of surface zoogloea (concavity) and the gas-driven collision and friction between granules which finally shaped the surface convexity. The produced dinitrogen gas links the metabolic activity with the formation of surface convexity and concavity of AnGS. This finding provided an alternative visible performance indictor of anammox process.

A secret of high-rate mass transfer in anammox granular sludge: "Lung-like breathing".

The granulation of anaerobic ammonium oxidation (Anammox) biomass can play a key role in developing stable and high-rate working of anammox process. It is important to know the working mechanism of anammox granular sludge (AnGS) for the optimization of reactor performance. In this study, a "lung-like breathing" determinator was invented to investigate the working behavior of AnGS in the bioreactor. The results showed that the AnGS had a regular expansion and contraction phenomenon, which was called "lung-like breathing". With the biological loading rate (BLR) at 0.114 kg-N/(kg-VSS·d), the expansion and contraction amplitude (ExCA) was 1.29 ±â€¯0.05%, and the expansion and contraction frequency (ExCF) was 39.3 ±â€¯1.6 times/h. The AnGS cultivated in a bioreactor with higher nitrogen removal rate (NRR) was found to have the higher ExCA and ExCF when determinated at the same BLR, and the "lung-like breathing" behavior of one type of AnGS was revealed to bear a significantly (p < 0.05) positive correlation with the specific anammox activity (SAA). The mass transfer flux from "lung-like breathing" was far greater than that from molecular diffusion, which was regarded as a vital mechanism for the AnGS to demonstrate its high activity. These findings provided theoretical basis and technical parameters for the optimization of anammox nitrogen removal process.

Color characterization of anammox granular sludge: Chromogenic substance, microbial succession and state indication.

Vision is a direct and convenient way to get information, and color characteristics are important visual information to identify objects. In this study, the L*a*b* color space was introduced for the first time to digitize the surface color of anammox granular sludge (AnGS). Three AnGSs under typical biological loading rates were chosen and the color value a*/b* was found to have a positive correlation with the specific anammox activity and the biological loading rate of AnGS. Cytochrome c was detected to be the key chromogenic substance determining the red hue of AnGS. The concentration of cytochrome c was highly consistent with the abundance of anammox bacteria. Nitrospira was observed to compete with Candidatus Kuenenia for nitrite under survival state, resulting in the obvious decrease of a* value; while the growth of sulfur-related Limnobacter and Thiobacillus was enhanced under starvation state, leading to the production of Fe-S compounds covering over the surface and decrease of a* and b* value simultaneously. The unique color characteristics of AnGS were evaluated as a visual indicator to serve the on-line control and better judgement of anammox process.

Characterization of oligotrophic AnAOB culture: morphological, physiological, and ecological features.

Anaerobic ammonium oxidation (anammox) process is regarded as a promising nitrogen removal technology to treat ammonium wastewaters in a wide concentration range. Oligotrophic anaerobic ammonia oxidation bacteria (O-AnAOB) culture has been successfully achieved from a new anammox system to treat superlow ammonium concentration wastewaters. In this work, the O-AnAOB culture was compared with the eutrophic AnAOB (E-AnAOB) culture to reveal its physiological, morphological, and ecological features. Results showed that the specific anammox activity (SAA) of O-AnAOB culture was 0.07 kgN/(kgVSS·d) with the nitrogen removal rate (NRR) of 0.20 kgN/ (m3 d) in the reactor, while the SAA of E-AnAOB culture was 2.11 kgN/(kgVSS·d) with the NRR of 11.10 kgN/(m3 d). The hzs gene transcription levels (hzs-mRNA) of O-AnAOB and E-AnAOB cultures were 1.32 × 109 copies/gVSS and 1.51 × 1010 copies/gVSS, respectively. Morphologically, the O-AnAOB culture took on the unique brown color rather than the typical red color of E-AnAOB. The O-AnAOB cells lived in a disperse pattern in the culture. The cells were seriously deformed with deep craters on the cell wall. The size of anammoxsome and paryphoplasm compartments inside the O-AnAOB cells was smaller than that inside the E-AnAOB cells. Ecologically, the O-AnAOB culture had special microbial community with a higher bacterial diversity than the E-AnAOB. The most dominant genera in O-AnAOB were Anaerolineaceae (33.7%, fermentative bacteria), Candidatus Kuenenia (17.4%, anammox bacteria), and Nitrospira (7.3%, nitrite oxidizing bacteria). This study provided an insight into the new anammox process for deep nitrogen removal from wastewaters.

Nitrate removal and microbial analysis by combined micro-electrolysis and autotrophic denitrification.

A process combining micro-electrolysis and autotrophic denitrification (CEAD) with iron-carbon micro-electrolysis carriers was developed for nitrate removal. The process was performed using organic-free influent with a NO3(-)-N concentration of 40.0±3.0mg/L and provided an average nitrate removal efficiency of 95% in stable stages. The total nitrogen removal efficiency reached 75%, with 21% of NO3(-)-N converted into NH4(+)-N. The corresponding hydraulic retention time was 8-10h, and the optimal pH ranged from 8.5 to 9.5. Microbial analysis with high-throughput sequencing revealed that dominant microorganisms in the reactor belonged to the classes of ß-, γ-, and α-Proteobacteria. The abundance of the genera Thermomonas significantly increased during the operation, comprising 21.4% and 24.1% in sludge attached to the carriers in the middle and at the bottom of the reactor, respectively. The developed CEAD achieved efficient nitrate removal from water without organics, which is suitable for practical application.