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1. Bimetallic hybrid nanoflowers containing l-aspartic acid were synthesized by a one-pot method for successful immobilizing snailase.2. Asp@ZIF-ZnCo-Sna has excellent enzyme loading capacity, activity and better catalytic efficiency.

Noncompressible hemorrhage caused by gunshots and sharp objects leads to higher trauma mortality, and cryogels have great potential in controlling noncompressible hemorrhage applications owing to their shape-memory properties. However, the use of non-toxic crosslinkers to prepare cryogels for noncompressible hemorrhage remains a challenge. In this study, a series of cryogels were prepared using oxidized dextran (ODex) as a biocompatible crosslinker, combined with the good hemostatic properties of chitosan (CS) and human-like collagen (HLC), and polydopamine nanoparticles (PDA-NPs) were also introduced to strengthen the shape recovery speed of the cryogels and further enhance their hemostatic performance. The CS/HLC/ODex/PDA-NPs (CHOP) cryogels presented a highly interconnected macroporous structure, powerful water/blood absorption capacity, robust mechanical performance, and rapid water/blood-triggered shape recovery. In vitro coagulation and coagulation mechanism tests showed that CHOP exhibited strong procoagulant ability, high adhesion to blood cells and fibrinogen, and the capacity to activate platelets and intrinsic pathways. In vivo hemostatic tests indicated that CHOP could effectively shorten the bleeding time and reduce the bleeding volume of liver incision bleeding and liver noncompressible hemorrhage. Meanwhile, CHOP exhibited good biocompatibility and biodegradability, and could promote wound healing. These results suggest that CHOP cryogels will be a promising hemostatic dressing.

Hepatocellular carcinoma (HCC) is one of the most prevalent and deadly cancers in the world. Recently, suppression of glutamine metabolism has become one of the hottest therapy targets for cancer treatment. There is a growing amount of research that indicates that ginsenosides possess good anti-tumor activity. However, the effect of ginsenoside Rk1 on glutamine metabolism in HCC is unclear. In this study, Rk1 was demonstrated to be effective at inhibiting the proliferation of HCC through the induction of cell cycle arrest and apoptosis. Especially, Rk1 was shown for the first time to inhibit glutamine metabolism in HCC. Rk1 downregulates GLS1 expression, and consequently decreases the GSH production, stimulating ROS accumulation to induce apoptosis. In addition, transcriptomic results showed that the ERK/c-Myc signaling pathway was enriched in HepG2. Rk1 exerts an inhibitory effect on glutamine metabolism in HCC by regulating the ERK/c-Myc signaling pathway, and inducing apoptosis

Ginsenoside CK alleviated non-alcoholic fatty liver disease through the LKB1/AMPK pathway.

Ginsenoside Rh4 inhibited inflammation, oxidative stress and apoptosis in mice with gastric ulcers through modulation of MAPK/NF-κB and PGE2-Cox pathways, thereby ameliorating gastric injury.

To investigate the potential of ginsenosides in treating osteoporosis, ginsenoside compound K (GCK) was selected to explore the potential targets and mechanism based on network pharmacology (NP). Based on text mining from public databases, 206 and 6590 targets were obtained for GCK and osteoporosis, respectively, in which 138 targets were identified as co-targets of GCK and osteoporosis using intersection analysis. Five central gene clusters and key genes (STAT3, PIK3R1, VEGFA, JAK2 and MAP2K1) were identified based on Molecular Complex Detection (MCODE) analysis through constructing a protein–protein interaction network using the STRING database. Gene Ontology (GO) analysis implied that phosphatidylinositol-related biological process, molecular modification and function may play an important role for GCK in the treatment of osteoporosis. Function and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis suggested that the c-Fms-mediated osteoclast differentiation pathway was one of the most important mechanisms for GCK in treating osteoporosis. Meanwhile, except for being identified as key targets based on cytoHubba analysis using Cytoscape software, MAPK and PI3K-related proteins were enriched in the downstream of the c-Fms-mediated osteoclast differentiation pathway. Molecular docking further confirmed that GCK could interact with the cavity on the surface of a c-Fms protein with the lowest binding energy (−8.27 Kcal/moL), and their complex was stabilized by hydrogen bonds (Thr578 (1.97 Å), Leu588 (2.02 Å, 2.18 Å), Ala590 (2.16 Å, 2.84 Å) and Cys 666 (1.93 Å)), van der Waals and alkyl hydrophobic interactions. Summarily, GCK could interfere with the occurrence and progress of osteoporosis through the c-Fms-mediated MAPK and PI3K signaling axis regulating osteoclast differentiation.

Depression is a neuropsychiatric disease that threatens the physical and mental health of humans worldwide. This study explored the potential anti-depressant effects of ginsenoside Rh4 and its mechanisms of action. The results showed that Rh4 could significantly inhibit depression-like behavior in the depression mouse model and alleviate neuronal damage and hypothalamic-pituitary-adrenal axis disorder. Concurrently, Rh4 inhibits hippocampal neuronal apoptosis and synaptic structural damage due to the overexpression of proinflammatory cytokines and overactivation of microglia and astrocytes by inhibiting the immune-inflammatory response and signaling molecular interaction pathways. Rh4 can also improve intestinal flora and increase the short-chain fatty-acids content. The correlation analysis indicated that the Rh4-inhibited LPS/NLRP3/caspase-1/IL-1β signaling pathway plays a key role in ameliorating depression. Therefore, this study provides valuable insights into the mode of action of Rh4 on the brain-gut axis in depression, suggesting that Rh4 may be a promising clinical drug for the treatment of depression.

The pH diversification has been proved as an important factor affecting the self-assembly of spidroin. Herein, we constructed a novel spider silk protein (NT-MaSp1s-CT) with the pH-dependent secondary structures, containing pH-sensitive N-terminal, C-terminal domains and a repeated core region with merely 191 amino acids. Then pH sensitivity of NT-MaSp1s-CT was detected at different pH conditions and NT-MaSp1s-CT displayed pH-dependent conformational transitions consistent with rational designed objective. Besides, the micelles theory was employed to inquiry the assembly mechanism of NT-MaSp1s-CT in high concentration spinning dope. As expected, NT-MaSp1s-CT protein can be spun into continuous and uniform fibers with the pH ranging from 2 to 11, which is the largest pH boundary for artificial spider silk formation, simplifying the assembly conditions and paving a broad path for spinning process. Moreover, the hemolysis and cytotoxicity of NT-MaSp1s-CT fibers were also determined and the novel fibers exhibit excellent biocompatibility, providing wider potential applications in the biomedical and pharmaceutical fields.

Ginsenoside compound K (CK) with a wide range of pharmacological activities has been widely used in the healthcare product industry. However, the application of CK is limited by low productivity and difficult separation. The purpose of this study is to convert ginsenoside Rb1 into CK by improving conversion efficiency in novel “green” reaction medium-deep eutectic solvent (DES). Talaromyces purpureogenus was selected from ginseng rhizosphere soil to produce β-glucosidase with high activity and purity to transform ginsenosides, and Mn2+ was found to be an enzyme promoter. Among the DES based on choline chloride as hydrogen-bond receptor, choline chloride:ethylene glycol (ChCl:EG = 2:1) was the most promising solvent in maintaining enzyme activity and stability. In the presence of 30% v/v ChCl:EG = 2:1, the half-life of β-glucosidase was increased by 96%, the solubility of F2 was increased by 120%, and CK yield was increased by 54% compared with those in the buffer. Fourier transform infrared, circular dichroism, and fluorescence spectroscopy confirmed that DES did not destroy the structure and conformation of β-glucosidase. In addition, 80.6% CK conversion was obtained at 60 °C, pH 4.5, 48 h and 8 mM Rb1, which provided a feasible method for efficiently producing CK.

Multifunctional inorganic nanomaterials have opened new avenues for cancer diagnosis and therapy. However, the difficulty to functionalize them has prevented their wider application, owing to the lack of active groups on their surfaces. Here we report a novel method to functionalize manganese whitlockite (Mn-WH) with polydopamine (PDA) and hairpin DNA (hpDNA) to improve the water stability and anti-cancer effects of Mn-WH nanoparticles (Mn-WH NPs). Compared to WH NPs, the Mn-WH@PDA-hpDNA NPs exhibit better water dispersibility, high drug loading capacity, excellent photothermal performance, stable MRI imaging ability, and outstanding chemo-photothermal synergistic therapeutic potential against tumors. After intratumoral injection in nude mice, the Mn-WH@PDA-hpDNA-DOX NPs promote complete tumor ablation upon exposure to 808 laser-irradiation. The nanoparticles showed no major side effects or toxicity. Thus, these results indicate that the Mn-WH@PDA-hpDNA-DOX NPs have excellent potential as anti-cancer agents, along with excellent magnetic resonance imaging (MRI) capabilities and the reported functionalization approach provides a novel and effective strategy for the surface functionalization of inorganic nanomaterials.

In this study, a temperature-sensitive molecularly imprinted polymer was prepared by using the bifunctional monomer with the critical phase transition characteristics. Infrared spectrometry, scanning electron microscopy, and specific surface area testing were used to characterize the polymers. Then, the recognizing properties of the polymers were studied. Based on the prepared smart polymers, an SPE-HPLC analytical method for the determination of quinolizidine alkaloids in the extracts of Sophora flavescens was established and verified. Finally, the smart polymers were applied to the enrichment of quinolizidine alkaloids in plant extracts. By changing the temperature and solvents of the solid phase extraction conditions, the extraction process can increase the concentration of quinolizidine alkaloids by 4.3 to 5.2 folds. The extraction process has mild conditions and less time consumption, avoiding the use of a large number of toxic reagents, which indicate that the extraction process are more efficient and environmentally friendly.

Preventing bacterial infection in situ and accelerating skin generation simultaneously are essentially important for wound healing. Herein, a mussel-inspired Ag nanozyme-based bilayer hydrogel is constructed to address the above concerns. The bilayer hydrogel is composed of a layer with large pores absorbing the wound exudate and allowing oxygen exchange and a layer with small pores keeping the wound moist and preventing bacterial invasion. Benefitting from the polydopamine (PDA) coating-reduced Ag nanoparticles (AgNPs), the hydrogel exhibits high near infrared (NIR) absorption at 808 nm to generate hyperthermia and NIR-enhanced peroxidase (POD-like) activity to produce hydroxyl radicals (•OH), which endows the hydrogel with excellent antibacterial properties when combined with the released Ag+. In addition, the hydrogel presents adhesiveness due to the catechol group on a PDA molecule. The in vivo test results demonstrate that the bilayer hydrogel can accelerate infected skin generation by facilitating collagen deposition, decreasing tumour necrosis factor-α secretion, and promoting vascular endothelial growth factor expression.

Acetaminophen (APAP)-induced acute liver injury (AIALI) is one of the most common causes of acute liver failure. Owing to the limitations of N-acetylcysteine (NAC), which is the only antidote currently used in clinical practice for APAP, there is a need to develop new therapies that can provide extensive protection against AIALI. Ginsenoside Rk3 is a rare ginsenoside extracted from Panax notoginseng and a previous study has reported its excellent hepatoprotective function. In this study, we explored the therapeutic potential of ginsenoside Rk3 in APAP-induced acute liver injury. We found that ginsenoside Rk3 could reduce APAP-induced hepatotoxicity by reducing serum alanine aminotransferase and aspartate aminotransferase activity and pathological damage to the liver. Moreover, ginsenoside Rk3 could inhibit APAP-induced liver inflammation and oxidative stress by inhibiting the production of oxidative molecules, increasing the production of antioxidant molecules, and reducing the infiltration of inflammatory cells and the production of pro-inflammatory cytokines. Further mechanistic investigations revealed that the therapeutic effect of ginsenoside Rk3 was mainly dependent on the continuous activation of autophagy. Chloroquine, an autophagy inhibitor, was found to inhibit these protective effects. Therefore, ginsenoside Rk3 shows potential as a novel hepatoprotective agent to prevent drug-induced liver injury.

As the most common sleep disorder, insomnia seriously affects people's everyday lives. Phytochemicals have been shown to have excellent sleep-promoting effects. Therefore, this study was designed to investigate whether Rg5 and Rk1 extracted from ginseng had sleep-promoting effects and to explore their potential mechanisms. The results showed that Rg5 and Rk1 could significantly lessen the locomotor activity of mice and promote the sleep quality index, including increasing the amount of sleep in a pentobarbital sodium experiment with a threshold dose. In parallel, Rg5 and Rk1 could significantly shorten the sleep latency of mice and prolong the sleep time of mice. Furthermore, Rg5 and Rk1 augmented the GABA/Glu ratio, up-regulating the expression of the GABAA receptor and the GABAB receptor, whereas the GABAA receptor antagonist picrotoxin could antagonize the sleep quality of Rg5/Rk1. In addition, 5-HTP, the precursor of 5-HT, could enhance the sleep effect of Rg5 and Rk1 in mice, and both Rg5 and Rk1 could up-regulate the expression of 5-HT1A. These results were also confirmed by the detection of GABA and 5-HT in mouse cecum content. In conclusion, ginsenoside Rg5/Rk1 can exert sedative and hypnotic effects by affecting the GABA nervous system and the serotonin nervous system.

Background and purpose Ginsenoside Rg5 (Rg5), a triterpene saponin, extracted from the natural herbal plant ginseng, is one of the most potent anticancer drugs against various carcinoma cells. However, the therapeutic potential of Rg5 is limited by its low solubility in water, poor bioavailability, and nontargeted delivery. Therefore, we prepared folic acid (FA)-modified bovine serum albumin (BSA) nanoparticles (FA-Rg5-BSA NPs) to improve the therapeutic efficacy and tumor targetability of Rg5. Methods Various aspects of the FA-Rg5-BSA NPs were characterized, including size, polydispersity, zeta potential, morphology, entrapment efficiency (EE), drug loading (DL), in vitro drug release, thermal stability, in vitro cytotoxicity, cell apoptosis, cellular uptake, in vivo antitumor effects and in vivo biodistribution imaging. Results The FA-Rg5-BSA NPs showed a particle size of 201.4 nm with a polydispersity index of 0.081, uniform spherical shape, and drug loading of 12.64±4.02%. The aqueous solution of FA-Rg5-BSA NPs had favorable stability for 8 weeks at 4°C. The FA-Rg5-BSA NPs dissolved under acidic conditions. Moreover, the Rg5-BSA NPs and FA-Rg5-BSA NPs had advanced anticancer activity compared with Rg5 in MCF-7 cells, while poor cytotoxicity was observed in L929 cells. The FA-Rg5-BSA NPs facilitated cellular uptake and induced apoptosis in MCF-7 cells. In addition, in an MCF-7 xenograft mouse model, the in vivo antitumor evaluation revealed that FA-Rg5-BSA NPs were more effective in inhibiting tumor growth than Rg5 and Rg5-BSA NPs. The in vivo real-time bioimaging study showed that the FA-Rg5-BSA NPs exhibited superior tumor accumulation ability. Conclusion The results suggested that FA-Rg5-BSA NPs could serve as a promising system to improve the antitumor effect of Rg5.

7α- and 7β-hydroxysteroid dehydrogenases (HSDHs) are key biocatalysts for the biotransformation of ursodeoxycholic acid (UDCA) from chenodeoxycholic acid (CDCA). Various researches focused on heterogeneously expressed engineering enzymes to epimerize CDCA for UDCA, however not yet applied to further industrial application. In this work, we present the large-scale production of UDCA from CDCA by 7α- and 7β-HSDH enzymatic synthesis. Engineering enzymes were both successfully heterologous overexpressed in Escherichia coli BL21, and the effect of the enzymatic synthesis was investigated. The mass analysis (MS), IR spectrum, 1H NMR and 13C NMR were used to characterize the product. 500-L fermentor fermentation strategy producing a stable supply of HSDH and large-scale production process of UDCA in dozens kilogram class enabled industrial application.

Alcoholic liver disease (ALD), as one of the most common diseases, has become a global threat to human health. The aim of this study was designed to investigate the hepatoprotective effects of ginsenoside Rk3 against ALD and to discover the potential mechanisms of these protective effects. Mice were intragastrically administered 50% alcohol and treated with ginsenoside Rk3 (25 and 50 mg/kg) once per day for 6 weeks. The results indicated that ginsenoside Rk3 promoted hepatic function through significant downgrading AST and ALT levels in the serum, attenuating oxidative stress, and restoring antioxidant balance in hepatic tissue. Additionally, ginsenoside Rk3 significantly reduced the expression of inflammatory cytokines, such as NF-κB, TNF-α, IL-6, and IL-1β in the mice. Furthermore, ginsenoside Rk3 supplementation significantly inhibited apoptotic protein expression in the liver. The present study clearly demonstrates that ginsenoside Rk3 exerts a protective effect against ALD-induced liver injury because of its antioxidant, anti-apoptotic, and anti-inflammatory activities. The findings from the present investigation show that ginsenoside Rk3 might be a promising candidate treatment agent against ALD.

Epidemiological studies have shown that hepatocellular carcinoma (HCC) is a main cause of tumor death worldwide. Accumulating data indicate that ginsenoside CK is an effective compound for preventing HCC growth and development. However, improvement of pharmaceutical effect of the ginsenoside CK is still needed. In our study, we performed acetylation of ginsenoside CK (CK-3) and investigated the antitumor effects of the derivative in vitro and in vivo. The cytotoxicity analysis revealed that compared with CK, CK-3 could inhibit the proliferation of multiple tumor cell lines at a lower concentration. Treating with CK-3 on HCC cells arrested cell cycle in G2/M phase and induced cell apoptosis through AO/EB staining, TUNEL analysis and flow cytometry. Meanwhile, CK-3 significantly inhibited tumor growth in an HCC xenograft model and showed no side effect on the function of the main organs. Mechanistically, whole transcriptome analysis revealed that the antitumor effect of CK-3 was involved in the Hippo signaling pathway. The immunoblotting and immunofluorescence results illustrated that CK-3 directly facilitated the phosphorylation of YAP1 and decreased the expression of the main transcription factor TEAD2 in HCC cell lines and tumor tissue sections. Collectively, our results demostrate the formation of a new derivative of ginsenoside CK and its regulatory mechanism in HCC, which could activate the Hippo-YAP1-TEAD2 signaling pathway to regulate HCC progression. This research could provide a new direction for traditional Chinese medicine in the therapy of tumors.

Ginsenoside Rh4, as a bioactive component obtained from Panax notoginseng, has excellent pharmacological properties. However, its role in regulating gut microbiota and intestinal inflammation is still poorly understood. Thus, the aim of this study is to investigate the effect of Rh4 on gut microbiota, especially antibiotic-induced microbiota perturbation, and the underlying mechanisms. C57BL/6 mice were given different doses of Rh4 after the establishment of a gut microbiota disturbance model with antibiotics. Our data revealed that Rh4 administration could greatly improve the pathological phenotype, gut barrier disruption, and intestinal inflammation in mice that had been antibiotic-induced. Notably, it was found that Rh4 significantly inhibited the TLR4-MyD88-MAPK signaling pathway. In addition, Rh4 treatment could significantly increase the number of short chain fatty acids (SCFAs) and bile acids (BAs). These changes were accompanied with beneficial alterations in gut microbiota diversity and composition. In conclusion, Rh4 improves intestinal inflammation and induces potentially beneficial changes in the gut microbiota, which are conducive to revealing host-microbe interactions.

Bacterial infection and oxidative stress remain critical problems for wound closure because they frequently trigger severe complications and delay wound healing. In addition, maintaining a moist microenvironment can promote skin regeneration. In this study, a bilayer hydrogel modified with tannic acid (TA) was constructed to accelerate wound repair. The bilayer hydrogel, composed of a layer with large pores to absorb the fluids and allow gas exchange and small pores to maintain the wound moist and prevent bacterial invasion, was initially developed. Thereafter, TA was introduced into the hydrogel to form a dual crosslinked network and endowed the hydrogel with adhesiveness, antibacterial, and oxidation resistance. In addition, the TA@bilayer hydrogel exhibited shape memory behaviour and self-healing ability due to the hydrogen bonds formed between TA and the bilayer hydrogel. As a result, the TA@bilayer hydrogel significantly promoted wound closure by accelerating collagen deposition, reducing tumour necrosis factor-α (TNF-α) levels, and facilitating the expression of vascular endothelial growth factor (VEGF).

Protopanaxatriol saponin (PPT) has excellent anti-cancer, anti-diabetes, and anti-anemia effects, but its effect on intestinal bacteria has rarely been studied. In this study, we investigated whether PPT has the ability to improve intestinal health in antibiotic-treated mice. Model mice were constructed using a broad-spectrum antibiotic, cephalosporin. The composition of the gut microbiota and relative concentration of short-chain fatty acids (SCFAs), short-chain fatty acid receptor proteins (GPR41, GPR43 and GPR109A), tight junction components (ZO-1 and occludin) and pro-inflammatory cytokines (TNF-α, IL-6, IL-1β, IL-22 and IFN-γ) were determined. The results showed that PPT improved the composition of the gut microbiota, increased the concentration of SCFAs as well as receptor proteins and tight junction proteins, and decreased the pro-inflammatory cytokines. These findings indicate that PPT has a protective effect on intestinal microbes and enhances the integrity of the intestinal barrier as well as alleviates colonic inflammation in antibiotic-treated mice.

Ginsenoside Rk3 (G-Rk3) is a main active ingredient of ginsenosides. Several recent studies demonstrated that ginsenosides have potential anti-type 2 diabetes mellitus (T2DM) properties. To evaluate the anti-T2DM effect of G-Rk3 and verify its potential mechanism, a high-fat-diet/streptozocin (HFD/STZ) induced model of T2DM in C57BL/6 mice and a high glucose induced insulin resistance model of HepG2 cells were applied in this research. Our analysis indicated that G-Rk3 reduced HFD/STZ induced hyperglycemia, and serum insulin and inflammation levels, and ameliorated glucose tolerance and insulin resistance, and prevented liver histological changes. Furthermore, it also significantly reduced lipid accumulation as shown by lower TG, LDL-C and TC serum concentrations and Oil Red O staining in liver tissues. The hypoglycemic effect of G-Rk3 seemed to be partially mediated via the inhibition of hepatic gluconeogenesis, which was supported by the activated p-Akt, p-FoxO1 and GLUT2 and inhibited FoxO1, PEPCK and G6pase protein expressions in the liver as well as increased glucose uptake in high glucose induced HepG2 cells. The gene expressions of hepatic gluconeogenesis were also down-regulated by G-Rk3 in HFD/STZ induced T2DM mice. In addition, G-Rk3 suppressed HFD/STZ induced lipid accumulation by regulating related gene and protein expressions such as p-ACC, FAS and SREBP-1, which are the downstream targets of AMPK. AMPK and Akt inhibitors significantly reversed G-Rk3 mediated hepatic gluconeogenesis and lipid accumulation. Thus, our study is the first to illustrate that G-Rk3 mediates hepatic gluconeogenesis and lipid accumulation via activating the AMPK/Akt signaling pathway in HFD/STZ induced T2DM mice.

Human-like collagen (HLC), the collagen produced using fermentation technology, has been demonstrated previously to promote wound healing. However, the healing property of HLC in gastric ulcers remains to be verified. In this study, we investigated the healing efficacy and healing mechanisms of HLC on gastric ulcers. To investigate whether HLC still has healing activity on gastric ulcers after gastric digestion, we simulated gastric digestion in vitro to obtain a human-like collagen digestion product (HLCP) and used it as the control drug. A chronic gastric ulcer model induced by 60% acetic acid in rats was used to evaluate the healing effect of gastric ulcers in this study. The results showed that oral administration of HLC or HLCP for 4 or 7 days promoted ulcer healing, which can be directly observed by significant reductions in ulcer area. The oral administration of HLC and HLCP significantly increased the protein expression of growth factors (EGF, HGF, VEGF, bFGF and TGF-β1) and the HGF receptor (HGFr), promoted collagen deposition, regulated the activity of NOS, and decreased pro-inflammatory cytokines (TNF-a, il-6, il-10) and endothelin-1 (ET-1) levels in gastric tissue. Moreover, cell experiments showed that the effects of HLC on cell proliferation and migration are mainly caused by its digestion products. These findings indicate that HLC may be used as a nutritional supplement or therapeutic drug to promote the healing of gastric ulcers.

CS–HLC–HA–β-GP (chitosan–human-like collagen–hyaluronic acid–β-sodium glycerophosphate) hydrogels were prepared based on the self-assembly of CS–HLC–HA (CCA) fibers. The effects of the fibers on the synthesis, characteristics and biomedical applications of CS–HLC–HA–β-GP (CCAG) hydrogels were studied for various HA contents. The synthesis mechanism of the novel CCAG hydrogel was explored using Fourier transform infrared spectroscopy (FT-IR) and X-ray photoelectron spectroscopy (XPS). The hydrogels were characterized by a swelling test, gelling time and enzymatic treatments. The results indicated that a new amide bond (–CONH) and –NRH2+ were formed. The gelling time and swelling behaviors were dependent on the intertwining, overlap and adsorption of the polymer chains at various temperatures and pH. Furthermore, biomedical applications were evaluated by transmission electron microscopy (TEM), immunohistochemical analysis and haematoxylin and eosin (H&E) staining. The effect of the fibers on the histocompatibility of the hydrogels revealed that the fibers inside the hydrogel pores reduced the quantity of macrophages, decreased the degree of inflammation, and improved the anti-degradation of the modified hydrogels. This type of new hydrogel emerges as an interesting injectable filling biomaterial for tissue engineering.

In this study, we designed multifunctionalized hydrogel scaffolds and injectable particles based on high-molecular-weight (MW) pullulan and human-like collagen (HLC) crosslinked with 1,4-butanediol diglycidyl ether (BDDE) for combination therapy tissue restoration. The properties of the pullulan/BDDE (PB) and pullulan/BDDE/human-like collagen (PBH) hydrogels were characterized via swelling ratio measurements, mechanical tests, and enzymatic degradation in vitro and via subcutaneous injections in vivo. The results demonstrate that the dry hydrogels completely returned to their original state in deionized water. The elastic modulus of the PBH53 dry hydrogels is higher than that of the other hydrogels after exposure to bending stress and compression stress with a maximum value of 7858.93 MPa. In addition, the in vitro live/dead staining and cell adhesion of the PBH hydrogels exhibited a superior fibroblast morphology without high levels of cell death, which were considerably better than those of PB hydrogels. In vivo, PB and PBH particles with good biocompatibility and anti-biodegradation were successfully prepared via the granulation of wet PB and PBH hydrogels for efficient subcutaneous injection in Kunming mice and New Zealand rabbits. Therefore, the PB and PBH hydrogels were found to be acceptable, safe, soft materials for use in skin restoration, cartilage treatment, and lacrimal dryness therapy.

Cartilage tissue engineering has become the trend of cartilage defect repair owing to the engineered biomimetic tissue that can mimic the structural, biological and functional characteristics of natural cartilage. Biomaterials with high biocompatibility and regeneration capacity are expected to be used in cartilage tissue engineering. Herein, in this study, a dual-network bovine serum albumin/sodium alginate with hydroxyapatite nanowires composite (B-S-H) hydrogel scaffold has been prepared for cartilage repair. The obtained B-S-H hydrogel scaffold exhibits ideal physical properties, such as excellent mechanical strength, high porosity and swelling ratio, as well as the excellent biological activity to promote the human bone marrow derived mesenchymal stem cells (hBMSCs) proliferation and differentiation. The in vivo study further shows that the B-S -H hydrogel scaffold can obviously promote the generation of new cartilage that integrates well with surrounding tissues and is similar to adjacent cartilage in terms of thickness. It is considered that the B-S-H hydrogel scaffold has great potential in the application of cartilage defects repair.

Lung adenocarcinoma (LAC) is a common lung cancer with a high malignancy that urgently needs to be treated with effective drugs. Ginsenoside Rh4 exhibits outstanding antitumor activities. However, few studies reported its effects on growth, metastasis and molecular mechanisms in LAC. Here, Rh4 is certified to show a strong anti-LAC efficiency in vitro and in vivo. Results of flow cytometry and Western blot are obtained to exhibited that Rh4 markedly restrained cellular proliferation and colony formation by arresting the cell cycle in the G1 phase. Results from a wound healing assay and transwell assays demonstrated that Rh4 is active in the antimigration and anti-invasion of LAC. The analysis of Western blot, immunofluorescence and RT-qPCR confirmed that Rh4 reverses the epithelial–mesenchymal transition (EMT) through upregulating the gene expression of E-cadherin and downregulating that of snail, N-cadherin and vimentin. In vivo results from immunohistochemistry show consistent trends with cellular studies. Furthermore, Rh4 suppresses the Janus kinases2/signal transducer and activator of the transcription3 (JAK2/STAT3) signaling pathway stimulated by TGF-β1. Silencing the STAT3 signal or co-treating with AG490 both enhanced the EMT attenuation caused by Rh4, which revealed that Rh4 suppressed EMT via inhibiting the JAK2/STAT3 signaling pathway. These findings explore the capacity and mechanism of Rh4 on the antimetastasis of LAC, providing evidence for Rh4 to LAC therapy.

In this study, fatty acid compositions, physiochemical properties, and antioxidant activities of eight varieties of kiwifruit seed oil (KSO) in China were comparatively investigated. All varieties of KSO were rich in unsaturated fatty acid (UFA), especially linolenic acid. Different varieties of KSO showed subtle differences in physico-chemical characteristics; e.g., density (0.88–0.091 mg/mL), refractive index (1.4803–1.4823), acid value (3.21–3.99 mg KOH/g oil), iodine value (177.22–202.75 g I/100 g oil), and peroxide value (7.23–14.75 mM/kg oil). Different varieties of KSO demonstrated different radical scavenging capacities for DPPH (IC50 = 25.7–35.1 mg/mL), HO (IC50=0.93–1.26 mg/mL), ORAC (1.26–1.99 mM Trolox/kg), and FRAP (3.30–132.30 mg Trolox/kg). Principal component analysis indicated the variety “Hongyang” to have a higher content of UFA and better antioxidant capacity than other varieties. Furthermore, comet assay verified that KSO can protect mice lymphocytes against oxidative DNA damage. Taken together, our findings provide evidence for the bioactivity of KSO as a potential dietary supplement.

Breast cancer is the most frequently diagnosed cancer and has become the main cause of cancer-related death among women worldwide. Traditional chemotherapy for breast cancer has serious side effects for patients, such as the first-line drug docetaxel. Ginsenoside Rg5, a rare ginsenoside and the main ingredient extracted from fine black ginseng, has been proved to have anti-breast cancer efficacy in vitro. Here, the in vivo anti-breast cancer efficacy, side effects and potential molecular mechanisms of Rg5 were investigated on a BALB/c nude mouse model of human breast cancer. The tumor growth inhibition rate of high dose Rg5 (20 mg kg-1) was 71.4 ± 9.4%, similar to that of the positive control docetaxel (72.0 ± 9.1%). Compared to docetaxel, Rg5 showed fewer side effects in the treatment of breast cancer. Treatment with Rg5 induced apoptosis and autophagy in breast cancer tissues. Rg5 was proved to induce caspase-dependent apoptosis via the activation of the extrinsic death receptor and intrinsic mitochondrial signaling pathways. The autophagy induction was related to the formation of an autophagosome and accumulation of LC3BII, P62 and critical Atg proteins. Further studies showed that Rg5 in a dose-dependent manner induced apoptosis and autophagy through the inhibition of the PI3K/Akt signaling pathway as indicated by the reduced phosphorylation level of PI3K and Akt. Taken together, Rg5 could be a novel and promising clinical antitumor drug targeting breast cancer.

Pyrolysis is potentially an effective treatment of oily sludge for oil recovery, and the addition of a catalyst is expected to affect its pyrolysis behavior. In the present study, Fe/Al-pillared bentonite with various Fe/Al ratios as pyrolysis catalyst is prepared and characterized by XRD, N2adsorption, and NH3-TPD. The integration of Al and Fe in the bentonite interlayers to form pillared clay is evidenced by increase in the basal spacing. As a result, a critical ratio of Fe/Al exists in the Fe/Al-pillared bentonite catalytic pyrolysis for oil recovery from the sludge. The oil yield increases with respect to increase in Fe/Al ratio of catalysts, then decreases with further increasing of Fe/Al ratio. The optimum oil yield using 2.0 wt% of Fe/Al 0.5-pillared bentonite as catalyst attains to 52.46% compared to 29.23% without catalyst addition in the present study. In addition, the addition of Fe/Al-pillared bentonite catalyst also improves the quality of pyrolysis-produced oil and promotes the formation of CH4. Fe/Al-pillared bentonite provides acid center in the inner surface, which is beneficial to the cracking reaction of oil molecules in pyrolysis process. The present work implies that Fe/Al-pillared bentonite as addictive holds great potential in industrial pyrolysis of oily sludge.

By combining DEAE-cellulose-52 and Sepharose-CL-6B chromatography, a heteropolysaccharide ECIP-1A with an average molecular weight of 24,336 Da was isolated from the hyphae of Eurotium cristatum. The main components of the monosaccharides of ECIP-1A were mannose, glucose and galactose, and their molar ratio is 4.003: 0.592: 4.58. Analysis of the immunomodulatory activity showed that ECIP-1A could significantly promote the proliferation of RAW264.7 cells and induce the production of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and nitric oxide (NO) by RAW264.7 cells. The results therefore suggested that ECIP-1A, as an immunomodulator, may have potential application prospects in the food and pharmaceutical industries.

Panax ginseng is a traditional Chinese medicine with significant pharmaceutical effects and broad application. Rare ginsenosides with high antitumor activities can be generated via oriented modification of their glycosyl moiety. For this purpose, suitable microorganisms and their enzymatic systems can be used. In this review, we address several issues associated with these systems. Under aerobic conditions, fungus biotransformation provides an efficient and inexpensive biotransformation process that can be easily scaled up. Considering the profound use of probiotics, wild strains generally recognized as safe have shown a potential through classical fermentation in food manufacturers of deglycosylated ginsenosides. Commonly applied recombinant enzymes from E. coli, especially recombinant hyperthermophilic enzymes, showed efficient conversion in biomedical or pharmaceutical industries. In this review, key genes dedicated to the production of ginsenosides (especially in Saccharomyces cerevisiae) are highlighted in relation to the large-scale production of ginsenosides. We also evaluate biocatalytic strategies that are aimed to improve product specificity and biocatalytic efficiency with industrial applications. Perspectives of protein engineering and solvent engineering in the development and large-scale preparation of ginsenosides in anticancer drugs, food and health care products are explored. KEY POINTS : • Modification of ginsenosides with food/engineered microorganisms is summarized. • Optimization of cell factories by protein engineering remains challenging. • Solvent engineering offers an attractive potential alternative.

Skin defects caused by various reasons are currently common clinical problems. At present, hydrogels have been proposed as tissue-engineered skin scaffolds to regenerate the tissues of the defect. We used human-like collagen (HLC), which was isolated and purified after high-density fermentation of recombinant E. coli BL21 The gel uses HLC and carboxymethylated chitosan (CCS) as raw materials and combines enzyme-chemical double cross-linking technology to form a three-dimensional porous network structure that mimics the human extracellular matrix, providing attachment points and nutrients for cell growth and proliferation. For comparison, we used a common hydrogel raw material, gelatin, to prepare a hydrogel in the same way. The experimental results show that the HLC-CCS skin scaffold hydrogel has good mechanical properties, high porosity and good histocompatibility. And full-thickness skin defect repair experiments show that this hydrogel has a good ability to promote skin tissue regeneration at the wound. In summary, this HLC-based double-crosslinked hydrogel can be used as a project strategy for skin defect repair.

Ginsenoside Rk1 is a substance derived from ginseng and exhibits various activities such as anti-diabetic, anti-inflammatory and anti-cancer effects; however, its anti-tumor effect and target signaling mechanism in lung adenocarcinoma are not well understood. Here, we show that Rk1, a natural drug product, can function as an antitumor modulator that induces apoptosis in lung adenocarcinoma cells by inhibiting NF-κB transcription and triggering cell cycle arrest. Mechanistically, Rk1 suppressed the proliferation and clonal formation of two lung adenocarcinoma cell lines (A549 and PC9) in vitro and caused G1 phase cell arrest. In the A549 xenograft model, Rk1 significantly inhibited tumor growth and had few toxic side effects on normal organs. Western blotting results showed that Rk1 increased the protein expression of Bax, cleaved caspase-3, -8, and -9, and PARP, decreased the expression of Bcl-2 and blocked the NF-κB signaling pathway. Furthermore, ginsenoside Rk1 also reduced the high expression of PD-L1 in lung adenocarcinoma cells by inhibiting NF-κB signaling. These data revealed a previously unreported antitumor mechanism of Rk1, providing new ideas and an experimental basis for further study of the mechanism of action of Rk1 in lung adenocarcinoma.

Ginsenoside Rb1 is the main predominant component in Panax species. In this study, an eco-friendly and convenient preparation method for ginsenoside CK has been established, and five strains of β-glucosidase-producing microorganisms were screened out from the soil of a Panax notoginseng planting field using Esculin-R2A agar. Aspergillus niger XD101 showed that it has excellent biocatalytic activity for ginsenosides; one of the isolates can convert ginsenoside Rb1 to CK using extracellular enzyme from the mycelium. Mycelia of A. niger were cultivated in wheat bran media at 30 °C for 11 days. By the removal of mycelia from cultured broth, enzyme salt fractionation by ammonium sulfate (70%, v/v) precipitation, and dialysis, sequentially, crude enzyme preparations from fermentation liquid supernatant were obtained. The enzymatic transformed Rb1 as the following pathways: Rb1→Rd→F2→CK. The optimized reaction conditions are at reaction time of 72 h, in the range of pH 4–5, and temperature of 50–60 °C. Active minor ginsenosides can be obtained by a specific bioconversion via A. niger XD101 producing the ginsenoside-hydrolyzing β-glucosidase. In addition, the crude enzyme can be resulted in producing ginsenoside CK via conversion of ginsenoside Rb1 at high conversion yield (94.4%). FDA generally regarded, A.niger as safe microorganism. Therefore, these results indicate that A. niger XD10 may provide an alternative method to prepare ginsenoside CK without food safety issues in the pharmaceutical industry.

In this study, we fabricated multifunctionalized hydrogel scaffolds based on hyaluronic acid (HA)-tyrosine and human-like collagen (HLC) by crosslinking with 1, 4-butanedioldiglycidyl ether (BDDE) for soft tissue fillers. The physicochemical of HA-tyrosine/BDDE (HTB) and various proportions of HA-tyrosine/BDDE/HLC (HTBH) hydrogels were characterized by swelling ratio, mechanical strength, morphology and thermal stability. The results demonstrated various HTBH hydrogels had superior performance in mechanical properties than HTB hydrogels. The biodegradation in vitro results demonstrated the degradation of HTBH hydrogels were slower than HTB hydrogels, and residual masses of HTBH hydrogels varied with different proportions of HA-tyrosine and HLC in the presence of the combination of HAse/collagenase I. The cell counting kit-8 (CCK-8), Hoechst/PI staining and cell adhesion of various HTBH hydrogels showed less cytotoxic and superior fibroblast attachment than those of HTB hydrogels. Moreover, subcutaneous injections of HTBH hydrogels containing high proportions of HLC in mice and rabbits all exhibit better biocompatibility and anti-biodegradation compared to HTB hydrogels after 1, 4, 8, 12 and 16 weeks. Hematoxylin-eosin staining (H&E), immunohistochemical and transmission electron microscope (TEM) analysis results indicated HTBH hydrogels had less intense inflammatory responses with increase proportion of HLC. Taken together, HTBH hydrogels had great potential application as ideal soft tissue fillers with excellent mechanical properties, biological stability and biocompatibility.

The purpose of this study was to clearly understand the health-promoting potentials of broccoli seeds and sprouts according to identify their representative bioactive compounds and antioxidant activities in six varieties. Sulforaphane (SF) extraction was firstly optimized from seeds and sprouts. Then SF extracted under optimized conditions from seeds and sprouts were compared. Most varieties obtained the maximum SF, total phenolic (TP) and flavonoid (TF) contents in sprouts on day 3. SF contents in sprouts were 46%–97% of seeds, whereas TP and TF contents in sprouts were 1.12–3.58 times higher than seeds among varieties. After in vitro digestion, broccoli sprouts from MNL variety kept considerable SF, TF, and TP contents, as well as antioxidant capacities, with all values higher than seeds. Compared with seeds, sprouts after 3 days germination were also recommended as raw materials of functional foods that possess high health-promoting potential.

This studyinvestigated the behavior and kinetics of co-pyrolysis of apple wood (AW)with H3PO4and K3PO4as catalysts under microwaveto prepare biochar as microbialabsorbent. The kinetic studies indicate that the co-pyrolysis of AW withH3PO4orK3PO4can effectively improve the pyrolysis efficiencyand enhance the biocharcharacteristicsby reducing ofthe activation energy of the pyrolysis reaction. The kinetic parameters indicate that the activation energy of the mixturesin the main pyrolysis stage is lower than that of a single AW, whichmeanthat the co-pyrolysis of AW withH3PO4orK3PO4shows excellent synergy. Biochar characterization showed that the yield of biochar reachedthe highest58.6% whenthe ratio(H3PO4/AW) is0.5. The adsorption results show that the bacteria SL-44 can be effectively loaded on the surface of the biochar, and the adsorption process is combined with Langmuir model and process can proceed spontaneously.

This study involved the development of a β-glucosidase-catalyzed hydrolysis method based on a deep eutectic solvent (DES), choline chloride-ethylene glycol 2:1, and continuous feed technique to overcome the difficulty of high-concentration ginsenoside hydrolysis. A productivity of 142 mg·L−1·h−1 was achieved with the following conditions: 30 vol% DES, pH 5.0, 55 °C, and substrate concentration of 12 mM. In the presence of DES, the affinity and catalytic efficiency of β-glucosidase to Rd increased by 49 and 64%, respectively, which promoted the continuation of hydrolysis. Moreover, conformation of β-glucosidase was mostly retained, as confirmed by spectral information. Through a combination of a substrate fed-batch technique to reduce the inhibitory effects of substrates and products, the CK conversion rate increased by 44% compared to traditional single-batch in pure buffer. This report describes a practical method for the continuous conversion of natural compounds through biological processes and solvent engineering.

Obesity is considered as a chronic disease which seriously affecting people's health and daily life. Kiwifruit (Actinidia chinensis Planch) seed oil (KSO), as a by-product of kiwifruit processing, is rich in fatty acids. Conventional wisdom holds that KSO has many health benefits, but there is no scientific basis. Here, the relieving effects of KSO on obesity and its potential mechanism were investigated in high-fat diet (HFD)-induced C57BL/6 mice. Mice were divided into four groups: ND (normal diet); HFD; L-KSO and H-KSO (HFD supplemented with 1.0 and 3.0 mL/kg·bw of KSO per day, respectively). Results showed that continuous supplementation KSO for 12 weeks significantly decreased bodyweight, inguinal fat tissue weight, blood glucose, and HOMA-IR index and ameliorated serum lipids accumulation (TC, TG, HDL-C, and LDL-C). Relative mRNA expression of inflammatory cytokines (TNF-α, IL-6, IL-1β, COX-2, and iNOS) was down-regulated and expression of thermogenesis-related genes (PPAR-γ, UCP1, PGC1-α, and PRDM16) was up-regulated in the inguinal fat tissue of KSO treated mice. Principal component analysis showed that the microbial community compositions of four groups were different. KSO supplementation dramatically decreased the Firmicutes-to-Bacteroidetes ratio. Together, our findings demonstrated that long-term supplementation KSO ameliorates obesity by reducing inflammation, adipose thermogenesis and gut microbiota dysbiosis.

Ginsenoside Rk3 (Rk3) and ginsenoside Rh4 (Rh4) have been reported to have therapeutic effects on a variety of diseases including some hematological diseases. The aim of the present study was to treat ribavirin-induced anemia in mice with Rk3 and Rh4 and explore the therapeutic mechanisms. All the experimental mice were randomly divided into 10 groups (the normal group, the control group, the model group, the erythropoietin (EPO) group, and 6 groups of different doses of Rk3 and Rh4, n = 10 per group). The changes in the hemograms, the content of cytokines, the number of hematopoietic cells and viscera damage were observed in the experiment. The results showed that the red blood cell count, the hemoglobin amount and the hematocrit of mice in the ginsenoside groups could be restored to normal values in a short time, and the effects of both Rk3 and Rh4 are obviously better than those of EPO. Moreover, some positive regulators (EPO, erythroid transcription factor-1, and interleukin-3) related to hematopoiesis increased and some negative regulators (interferon-γ and tumour necrosis factor-α) decreased in vivo after the injection of Rk3 or Rh4. These two ginsenosides did little damage to viscera and produced different degrees of increase in the hematopoietic cells in the bone marrow and spleen, which indicated that Rk3 promoted bone marrow and extramedullary haematopoiesis while Rh4 promoted extramedullary hematopoiesis. Therefore, both Rk3 and Rh4 could be considered new therapeutic drugs for treating anemia.

Type III collagen is one of the most abundant proteins in the human body, which forms collagen fibrils and provides the stiff, resilient characteristics of many tissues. In this paper, a new method for secretory expression of recombinant hydroxylated human collagen α1(III) chain in Pichia pastoris GS115 was applied. The gene encoding for full-length human collagen α1(III) chain (COL3A1) without N-terminal propeptide and C-terminal propeptide was cloned in the pPIC9K expression vector. The prolyl 4-hydroxylase (P4H, EC 1.14.11.2) α-subunit (P4Hα) and β-subunit (P4Hβ) genes were cloned in the same expression vector, pPICZB. Fluorogenic quantitative PCR indicates that COL3A1 and P4H genes have been expressed in mRNA level. SDS-PAGE shows that secretory expression of recombinant human collagen α1(III) chain was successfully achieved in P. pastoris GS115. In addition, the result of amino acids composition analysis shows that the recombinant human collagen α1(III) chain contains hydroxyproline by coexpression with the P4H. Furthermore, liquid chromatography coupled with tandem mass spectrometry analysis demonstrates that proline residues of the recombinant human collagen α1(III) chain were hydroxylated in the X or Y positions of Gly-X-Y triplets.

Recombinant collagen and gelatin have been applied in biomedical materials field because of products from genetically engineered microorganisms with improved safety, traceability, reproducibility, and homogeneous quality. To obtain high-level secretory expression of single-chain full-length human collagen α1(III) chain (COL3A1) without the N and C telopeptides, the cDNA coding for the human COL3A1 gene was cloned into the secretory expression vector pPIC9K and integrated into Pichia pastoris GS115. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting analysis of culture supernatant from the recombinant methylotrophic yeast suggested that the unhydroxylated recombinant human COL3A1 (rhCOL3A1) was secreted into the culture medium, and exhibited an apparent molecular mass of approximately 130 kDa, which is 1.4 times higher than the theoretical one. Finally, the unhydroxylated rhCOL3A1 was purified to greater than 90% purity using a four-step approach. In addition, methylthiazolydiphenyl-tetrazolium bromide experiments indicated that low concentration of rhCOL3A1 could promote Baby hamster kidney cell (BHK21) proliferation effectively. The production and purification of rhCOL3A1 described in this study offer a new method for obtaining high level of rhCOL3A1 in relatively pure form, which is suitable for biomedical materials application.

Ginsenoside Rk3 (Rk3) is present in the roots of processed Panax notoginseng herbs and it exerts anti-platelet aggregation, pro-immunogenic and cardioprotective effects. However, little is known regarding the anticancer activities of this compound, especially in lung cancer. This study was designed to investigate the anticancer effects of Rk3 on non-small cell lung cancer (NSCLC) cells and in an H460 xenograft tumor model. Our results showed that Rk3 reduced cell viability, inhibited both cell proliferation and colony formation, and induced G1 phase cell cycle arrest by downregulating the expression of cyclin D1 and CDK4 and upregulating the expression of P21. Rk3 also induced apoptosis in a concentration-dependent manner in H460 and A549 cells by Annexin V/PI staining, TUNEL assay and JC-1 staining, resulting in a change in the nuclear morphology. Moreover, Rk3 induced the activation of caspase-8, -9, and -3, promoted changes in mitochondrial membrane potential, decreased the expression of Bcl-2, increased the expression of Bax, and caused the release of cytochrome c, which indicated that the apoptosis-inducing effects of Rk3 were triggered via death receptor-mediated mitochondria-dependent pathways. Furthermore, Rk3 significantly inhibited the growth of H460 xenograft tumors without an obvious effect on the body weight of the treated mice. Histological analysis indicated that Rk3 inhibited tumor growth by altering the proliferation and morphology of tumor cells. In addition, we confirmed that Rk3 inhibited angiogenesis via CD34 staining and chick embryo chorioallantoic membrane (CAM) assay in vivo. Taken together, our findings revealed not only the anticancer effect of Rk3 on NSCLC cells but also a new promising therapeutic agent for human NSCLC.

Collagen has been proven to be a valuable biomedical material for many medical applications. Human-like collagen (HLC) is a novel important biomedical material with diverse medical applications. In this work, recombinant Escherichia coli BL21 3.7 ∆ptsG was constructed, the characters of ptsG mutant strain were analyzed, and real-time quantitative polymerase chain reaction (PCR) was applied to investigate the effect of ptsG gene deletion on the transcriptional level of the phosphotransferase system (PTS) genes responsible for glucose transport. The HLC production and cell growth ability were 1.33- and 1.24-fold higher than those of its parent strain in the fermentation medium, respectively, and 1.16- and 1.17-fold in the modified minimal medium individually. The acetate accumulation decreased by 42%-56% compared to its parent strain in the fermentation medium, and 70%-87% in the modified minimal medium. The results of RT-qPCR showed that the transcriptional level of crr, ptsH, ptsI, and blgF in ptsG mutant all decreased dramatically, which inferred a decrease in the glucose uptake rate, but the transcriptional level of FruB and manX increased slightly, which demonstrated the activation of fructose- and mannose-specific transport pathways in the ptsG mutant. This study demonstrates that ptsG deletion is an effective strategy to reduce acetate accumulation and increase biomass and HLC production.