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Cellulose is one of the most abundant renewable resources in nature. However, its recalcitrant crystalline structure hinders efficient enzymatic depolymerization. Unlike cellulases, lytic polysaccharide monooxygenases (LPMOs) can oxidatively cleave glycosidic bonds in the crystalline regions of cellulose, playing a crucial role in its enzymatic depolymerization. An AA9 LPMO from Myceliophthora thermophila was previously identified and shown to exhibit a highly efficient catalytic performance. To further enhance its catalytic efficiency, consensus mutagenesis was applied. Compared with the wild-type enzyme, the oxidative activities of mutants A165S and P167N increased by 1.8-fold and 1.4-fold, respectively, and their catalytic efficiencies (kcat/Km) improved by 1.6-fold and 1.2-fold, respectively. The mutants also showed significantly enhanced activity in the synergistic degradation of cellulose with cellobiohydrolase. Additionally, the P167N mutant exhibited better H2O2 tolerance. A molecular dynamics analysis revealed that the increased activity of mutants A165S and P167N was due to the closer proximity of the active center to the substrate post-mutation. This study demonstrates that selecting appropriate mutation sites via a semi-rational design can significantly improve LPMO activity, providing valuable insights for the protein engineering of similar enzymes. [ABSTRACT FROM AUTHOR]

In this study, KGMOS (DP, 2-13), derived from KGM (Konjac glucomannan), was applied to elucidate plant immunity in a Nicotiana benthamiana Phytophthora nicotianae model. Application of KGMOS (25–100 mg/L) notably inhibited P. nicotianae, resulting in reduced disease indices and a significant accumulation of defense molecules such as H2O2 and callose. Transcriptomic analysis revealed that genes shared between KGMOS-treated and control plants are involved in signaling pathways, transcription regulation, hydrogen peroxide catabolism, and oxidative stress response. This suggests that KGMOS triggers H2O2 accumulation, callose deposition, and activation of the salicylic acid (SA) and jasmonic acid/ethylene (JA/ET) pathways after pathogen inoculation. Upregulated defense-response genes in the KGMOS group included SA-related late blight-resistant, pathogenesis-related (PR), and JA/ET-related ethylene response factor (ERF) genes. Heatmap analysis showed more upregulated defense genes (PR and NPR) related to SA in the KGMOS-treated group than in controls. RT-qPCR validation revealed significant upregulation of SA and JA/ET pathway genes in KGMOS-treated plants. Higher SA content in these plants suggests enhanced disease resistance. This study concludes that KGMOS pre-treatment induced resistance against P. nicotianae, especially at a lower concentration (25 mg/L). These findings could offer valuable insights for the future application of KGMOS in controlling plant diseases for sustainable agriculture and postharvest management. [ABSTRACT FROM AUTHOR]

Konjac glucomannan (KGM) is a natural polysaccharide derived from konjac, which has been widely used in various fields due to its numerous beneficial properties. However, the high viscosity and water absorption of KGM limit its application. Compared with KGM, Konjac glucomannan oligosaccharides (KGMOS) have higher water solubility and stronger application value. In this paper, a novel mannanase KgManA was cloned from Klebsiella grimontii to develop a new KGMOS-producing enzyme. Bioinformatic analysis shows that the structural similarity between KgManA and other enzymes was less than 18.33%. Phylogenetic analysis shows that KgManA shares different branches with the traditional mannanases containing the CMB35 domain, indicating that it is a novel mannanase. Then, the enzymatic properties were determined and substrate specificity was characterized. Surprisingly, KgManA is stable in a very wide pH range of 3.0 to 10.0; it has a special substrate specificity and seems to be active only for mannans without galactose in the side chain. Additionally, the three-dimensional structure of the enzyme was simulated and molecular docking of the mannotetraose substrate was performed. As far as we know, this is the first report to characterize the enzymatic properties and to simulate the structure of mannanase from K. grimontii. This work will contribute to the development and characterization of novel K. grimontii-derived mannanases. The above results indicate that KgManA is a promising tool for the production of KGMOS. [ABSTRACT FROM AUTHOR]

SUMMARY: As an essential enzyme in the uridine diphosphate (UDP)‐GlcNAc biosynthesis pathway, the significant role of N‐acetylglucosamine phosphate mutase (AGM) remains unknown in plants. In the present study, a functional plant AGM (AtAGM) was identified from Arabidopsis thaliana. AtAGM catalyzes the isomerization of GlcNAc‐1‐P and GlcNAc‐6‐P, and has broad catalytic activity on different phosphohexoses. UDP‐GlcNAc contents were significantly decreased in AtAGM T‐DNA insertional mutants, which caused temperature‐dependent growth defects in seedlings and vigorous growth in adult plants. Further analysis revealed that protein O‐GlcNAcylation but not N‐glycosylation was dramatically impaired in Atagm mutants due to UDP‐GlcNAc shortage. Combined with the results from O‐GlcNAcylation or N‐glycosylation deficient mutants, and O‐GlcNAcase inhibitor all suggested that protein O‐GlcNAcylation impairment mainly leads to the phenotypic variations of Atagm plants. In conclusion, based on the essential role in UDP‐GlcNAc biosynthesis, AtAGM is important for plant growth mainly via protein O‐GlcNAcylation‐level regulation. [ABSTRACT FROM AUTHOR]

Oligogalacturonides (OGs) are a bioactive carbohydrate derived from homogalacturonan. The OGs synthesized in this study significantly inhibited the mycelial growth of Rhizoctonia solani AG-4HGI in vitro, even at a low concentration (10 mg/L). The seed vigor test demonstrated that the application of 50 mg/L OGs to sugar beet seeds significantly increased average germination percentage, germination energy, germination index, and seedling vigor index. The same concentration of OGs also improved the seedling emergence percentage of sugar beet when seeds were sown in soil inoculated with D2 and D31 isolates, respectively. The lesion diameter on mature sugar beet roots caused by R. solani AG-4HGI isolates D2 and D31 also decreased by 40.60% and 39.86%, respectively, in sugar beets roots first treated with 50 mg/mL OGs in the wound site, relative to lesion size in untreated/pathogen inoculated wounds. Sugar beet roots treated with 50 mg/mL OGs prior to inoculation with the D2 isolate exhibited up-regulation of the defense-related genes glutathione peroxidase (GPX) and superoxide dismutase (SOD) by 2.4- and 1.6-fold, respectively, relative to control roots. Sugar beet roots treated with 50 mg/mL OGs prior to inoculation with D31 exhibited a 2.0- and 1.6-fold up-regulation of GPX and SOD, respectively, relative to the control. Our results indicate that OGs have the potential to be used for the protection of sugar beet against R. solani AG-4HGI. [ABSTRACT FROM AUTHOR]

Consider a multi-cell mobile edge computing network, in which each user wishes to compute the product of a user-generated data matrix with a network-stored matrix. This is done through task offloading by means of input uploading, distributed computing at edge nodes (ENs), and output downloading. Task offloading may suffer long delay since servers at some ENs may be straggling due to random computation time, and wireless channels may experience severe fading and interference. This paper aims to investigate the interplay among upload, computation, and download latencies during the offloading process in the high signal-to-noise ratio regime from an information-theoretic perspective. A policy based on cascaded coded computing and on coordinated and cooperative interference management in uplink and downlink is proposed and proved to be approximately optimal for a sufficiently large upload time. By investing more time in uplink transmission, the policy creates data redundancy at the ENs, which can reduce the computation time, by enabling the use of coded computing, as well as the download time via transmitter cooperation. Moreover, the policy allows computation time to be traded for download time. Numerical examples demonstrate that the proposed policy can improve over existing schemes by significantly reducing the end-to-end execution time. [ABSTRACT FROM AUTHOR]

Existing works on task offloading in mobile edge computing (MEC) networks often assume a task is executed once at a single edge node (EN). Downloading the computed result from the EN back to the mobile user may suffer long delay if the downlink channel experiences strong interference or deep fading. This paper exploits the idea of computation replication in MEC networks to speed up the downloading phase. Computation replication allows each user to offload its task to multiple ENs for repetitive execution so as to create multiple copies of the computed result at different ENs which can then enable transmission cooperation and hence reduce the communication latency for result downloading. Yet, computation replication may also increase the communication latency for task uploading, despite the obvious increase in computation load. The main contribution of this work is to characterize asymptotically an order-optimal upload-download communication latency pair for a given computation load in a multi-user multi-server MEC network. Analysis shows when the computation load increases within a certain range, the downloading time decreases in an inversely proportional way if it is binary offloading or decreases linearly if it is partial offloading, both at the expense of linear increase in the uploading time. [ABSTRACT FROM AUTHOR]

Endo‐polygalacturonase II B (PgaB) from Aspergillus luchuensis was orthologous to endo‐polygalacturonase from Aspergillus niger with mutant sites Thr42Ser and Glu52Ala. Mature pgaB gene was cloned from the genomic DNA of A. luchuensis and secreted expressed with over 90% purity in Pichia Pastoris and reached 1.0 g/L after 144 hr culture. The recombinant PgaB was further purified by Ni‐NTA chromatography. Using polygalacturonic acid (PGA) as substrate, the optimal condition for PgaB activity was 40°C and pH 4.5, respectively. Km and Vmax of PgaB were 0.19 mmol/l and 103.58 μmol min−1 mg−1, respectively. The relative activity of PgaB remained more than 60% and 40% of maximum activity at 50 and 60°C for 7 hr. PgaB increased the light transmittance by 85% and showed high efficiency in juice clarification. The main product was galacturonic acid oligosaccharides with degrees of polymers (DP) 1–3. The PgaB is a potential pectinolytic enzyme in food industries. Practical applications: Endo‐polygalacturonase II B (PgaB) was identified from Aspergillus luchuensis, a filamentous fungus widely used in food and beverage fermentation in East Asia. PgaB still kept its most activity at 60°C for 7 hr. Polygalacturonic acid (PGA) can be digested effectively by the PgaB and the main products are galacturonic acid oligosaccharides with degrees of polymers (DP) 1–3. PgaB shows high efficiency in juice clarification. The PgaB is a potential pectinolytic enzyme for the applications in food industries. [ABSTRACT FROM AUTHOR]

Colon cancer (CC) is the third common neoplasm worldwide, and it is still a big challenge for exploring new effective medicine for treating CC. Natural product promoting human health has become a hot topic and attracted many researchers recently. Pectin, a complex polysaccharide in plant cell wall, mainly consists of four major types of polysaccharides: homogalacturonan, xylogalacturonan, rhamnogalacturonan I and II, all of which can be degraded into various pectin oligosaccharides (POS) and may provide abundant resource for exploring potential anticancer drugs. POS have been regarded as a novel class of potential functional food with multiple health-promoting properties. POS have antibacterial activities against some aggressive and recurrent bacterial infection and exert beneficial immunomodulation for controlling CC risk. However, the molecular functional role of POS in the prevention of CC risk and progression remains doubtful. The review focuses on antioxidant and anti-inflammatory roles of POS for promoting human health by regulating some potential oxidative and inflammation-activated pathways, such as ATP-activated protein kinase (AMPK), nuclear factor erythroid-2-related factor-2 (Nrf2), and nuclear factor-κB (NF-κB) pathways. The activation of these signaling pathways increases the antioxidant and antiinflammatory activities, which will result in the apoptosis of CC cells or in the prevention of CC risk and progression. Thus, POS may inhibit CC development by affecting antioxidant and antiinflammatory signaling pathways AMPK, Nrf2, and NF-κB. However, POS also can activate signal transduction and transcriptional activator 1 and 3 signaling pathway, which will reduce antioxidant and anti-inflammatory properties and promote CC progression. Specific structural and structurally modified POS may be associated with their functions and should be deeply explored in the future. The present review paper lacks the important information for the linkage between the specific structure of POS and its function. To further explore the effects of prebiotic potential of POS and their derivatives on human immunomodulation in the prevention of CC, the specific POS with a certain degree of polymerization or purified polymers are highly demanded to be performed in clinical practice. [ABSTRACT FROM AUTHOR]

In this paper, we study the probabilistic caching for an $N$ -tier wireless heterogeneous network (HetNet) using stochastic geometry. A general and tractable expression of the successful delivery probability (SDP) is first derived. We then optimize the caching probabilities for maximizing the SDP in the high signal-to-noise ratio regime. The problem is proved to be convex and solved efficiently. We next establish an interesting connection between $N$ -tier HetNets and single-tier networks. Unlike the single-tier network where the optimal performance only depends on the cache size, the optimal performance of $N$ -tier HetNets depends also on the base station (BS) densities. The performance upper bound is, however, determined by an equivalent single-tier network. We further show that with uniform caching probabilities regardless of content popularities, to achieve a target SDP, the BS density of a tier can be reduced by increasing the cache size of the tier when the cache size is larger than a threshold; otherwise, the BS density and BS cache size can be increased simultaneously. It is also found analytically that the BS density of a tier is inverse to the BS cache size of the same tier and is linear to BS cache sizes of other tiers. [ABSTRACT FROM PUBLISHER]

As a large number of new energy is employed as the driving force for the operation and transportation machinery of underground space projects, the lithium battery load in confined spaces, such as working faces, roadways and tunnels increases in geometric progression, and the coupled risks of heat damage and smoke poisoning caused by possible fires become more serious. In this paper, experimental and numerical methods were implemented to study the propagation mechanism of heat- and mass-induced disasters under catastrophic conditions, and a quantitative characterization model of multiple risk factors of thermal runaway and toxic gas diffusion of battery fire was proposed. The fuzzy analytical hierarchy process (FAHP) was conducted to calculate and grade the risk of lithium battery fire in a typical mine working face under multiple factors, including hazard source, personnel, working environment and emergency response. In addition, a quantitative early warning and control model was established for identified high-risk probability events. The results promote the quantitative and scientific development of multiple risk assessment and decision-making of confined space fire.

Actinidia arguta, commonly called hardy kiwifruit or kiwiberry, is a perennial vine of Actinidiaceae Actinidia genus. Understanding the main pathogens that cause the fruit rot of A. arguta during storage is of great significance for finding strategies to prevent fruit rot. In September 2020, the A. arguta (Sieb.et Zucc.) Planch. ex Miq. LD133 was harvested from a farm in Dandong City, Liaoning Province, China (40°31'N, 124°20'E). After being stored at room temperature for about a week, the fruit rotted (no mechanical damage or wound). Initial symptoms were localized irregular spots, which then became soft and the spots connected into large, flaky, light brown lesions. Later symptoms were dark brown lesions and rot that affect the entire fruit. The strain that shows the same morphology as observed in the four decaying tissues was isolated on PDA. The strain was white, edge irregular and surface wavy, and the reverse side was pale yellow (Fig. S1A). The black viscous acervuli appeared on the surface of the mycelium after 7 to 10 days at 25℃ with 12 hours photoperiod. Conidia were fusiform to ellipsoid, straight to slightly curved, 5.7×25.9 µm (width × length), n=30, with five versicolor cells (three brown median cells, two hyaline cells on apical and basal). The apical cell generally contains one to four appendages on conidia (Fig. S1B). For identify the selected strains, three genetic regions (ITS, TUB and TEF 1-α) were used for amplification and sequencing. These sequences of pathogen shared 98 to 100% homology with Neopestalotiopsis clavispora. Then, a phylogenetic tree was constructed by the Bayesian algorithm using PhyloSuite (v1.2.2) (Zhang et al. 2020). Based on the morphological and molecular characterization, the pathogen was identified as Neopestalotiopsis clavispora (Chamorro et al. 2016). Next, pathogenicity of the screened strains was determined by wound inoculation method. Ripe healthy fruits are immersed in 1% NaClO, rinsed twice with sterile water. A sterile needle was used to penetrate 1-2mm of peel, and then inoculate hyphae (about 5mm in length, 1mm in diameter), and 10 µL of sterile water as a control. The treated fruits are stored in an artificial climate chamber (22°C, 70% relative humidity with 12 hours photoperiod). The inoculated fruit began to show signs of rot on the second day, and after 6 days, lesions similar to those found during storage of the pathogenic isolated fruit appeared. Similarly, 10 µL (106 conidia/mL) conidial suspension exhibited decay symptoms. Compared with conidial suspension as inoculum, hyphae has shorter incubation period and stronger pathogenicity to fruit (Fig. S2). The pathogen was re-isolated from these infected fruits and identified as N. clavispora, thus fulfilling Koch's postulates. N. clavispora has been reported causing root and crown rot on strawberry in Spain, Argentina, Uruguay and Italy (Chamorro et al. 2016; Gilardi et al. 2019; Machin et al. 2019; Obregon et al. 2018), and as a pathogen on blueberry in Spain and Korea (Borrero et al. 2018; Lee et al. 2019), and causing leaf spot on macadamia in Brazil (Santos et al. 2019), and causing leaf spot on Syzygium cumini in India (Banerjee and Rana 2020). However, to our knowledge, this is the first report of Neopestalotiopsis clavispora causing postharvest fruit rot of A. argute in the worldwide. The identification of the pathogen is of great significance for conducting research on A. argute fruit preservation to prolong its shelf life and improve its merchantability.