1.Characterization of a D-mannitol oxidase from Paenibacillus sp. and its application in the preparation of D-mannose.
Ran LI ; Cong SONG ; Xiang ZHANG ; Zhenhua JIA
Chinese Journal of Biotechnology 2023;39(11):4682-4693
D-mannose has many functional activities and is widely used in food, medicine, agriculture and other industries. D-mannitol oxidase that can efficiently convert D-mannitol into D-mannose has potential application in the enzymatic preparation of D-mannose. A D-mannitol oxidase (PsOX) was found from Paenibacillus sp. HGF5. The similarity between PsOX and the D-mannitol oxidase (AldO) from Streptomyces coelicolor was 50.94%. The molecular weight of PsOX was about 47.4 kDa. A recombinant expression plasmid pET-28a-PsOX was constructed and expressed in Escherichia coli BL21(DE3). The Km and kcat/Km values of PsOX for D-mannitol were 5.6 mmol/L and 0.68 L/(s·mmol). Further characterization of PsOX showed its optimal pH and temperature were 7.0 and 35 ℃, respectively, while its enzyme activity could be stably remained below 60 ℃. The molar conversion rate of 400 mmol/L D-mannitol by PsOX was 95.2%. The whole cells of PsOX and AldO were used to catalyze 73 g/L D-mannitol respectively. The reaction catalyzed by PsOX completed in 9 h and 70 g/L D-mannose was produced. PsOX showed a higher catalytic efficiency compared to that of AldO. PsOX may facilitate the enzymatic preparation of D-mannose as a novel D-mannose oxidase.
Recombinant Proteins/metabolism*
;
Paenibacillus/metabolism*
;
Mannose/metabolism*
;
Escherichia coli/metabolism*
;
Mannitol/metabolism*
2.Zhizhu Decoction Alleviates Intestinal Barrier Damage via Regulating SIRT1/FoxO1 Signaling Pathway in Slow Transit Constipation Model Mice.
Yong WEN ; Yu ZHAN ; Shi-Yu TANG ; Fang LIU ; Qiu-Xiao WANG ; Peng-Fei KONG ; Xue-Gui TANG
Chinese journal of integrative medicine 2023;29(9):809-817
OBJECTIVE:
To explore the possible effects and mechanism of Zhizhu Decoction (ZZD) on the pathophysiology of slow transit constipation (STC).
METHODS:
A total of 54 C57BL/6 mice was randomly divided into the following 6 groups by a random number table, including control, STC model (model), positive control, and low-, medium- and high-doses ZZD treatment groups (5, 10, 20 g/kg, namely L, M-, and H-ZZD, respectively), 9 mice in each group. Following 2-week treatment, intestinal transport rate (ITR) and fecal water content were determined, and blood and colon tissue samples were collected. Hematoxylin-eosin and periodic acid-Schiff staining were performed to evaluate the morphology of colon tissues and calculate the number of goblet cells. To determine intestinal permeability, serum levels of lipopolysaccharide (LPS), low-density lipoprotein (LDL) and mannose were measured using enzyme-linked immunosorbent assay (ELISA). Western blot analysis was carried out to detect the expression levels of intestinal tight junction proteins zona-occludens-1 (ZO-1), claudin-1, occludin and recombinant mucin 2 (MUC2). The mRNA expression levels of inflammatory cytokines including tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, IL-4, IL-10 and IL-22 were determined using reverse transcription-quantitative reverse transcription reaction. Colon indexes of oxidative stress were measured by ELISA, and protein expression levels of colon silent information regulator 1/forkhead box O transcription factor 1 (SIRT1/FoxO1) antioxidant signaling pathway were detected by Western blot.
RESULTS:
Compared with the model group, ITR and fecal moisture were significantly enhanced in STC mice in the M-ZZD and H-ZZD groups (P<0.01). Additionally, ZZD treatment notably increased the thickness of mucosal and muscular tissue, elevated the number of goblet cells in the colon of STC mice, reduced the secretion levels of LPS, LDL and mannose, and upregulated ZO-1, claudin-1, occludin and MUC2 expressions in the colon in a dose-dependent manner, compared with the model group (P<0.05 or P<0.01). In addition, ZZD significantly attenuated intestinal inflammation and oxidative stress and activated the SIRT1/FoxO1 signaling pathway (P<0.05 or P<0.01).
CONCLUSION
ZZD exhibited beneficial effects on the intestinal system of STC mice and alleviated intestinal inflammation and oxidative stress via activating SIRT1/FoxO1 antioxidant signaling pathway in the colon.
Mice
;
Animals
;
Sirtuin 1/genetics*
;
Antioxidants
;
Occludin
;
Lipopolysaccharides
;
Claudin-1
;
Mannose
;
Mice, Inbred C57BL
;
Constipation/drug therapy*
;
Inflammation
;
Signal Transduction
3.Mannose inhibits the growth of prostate cancer through a mitochondrial mechanism.
Yu-Lin DENG ; Ren LIU ; Zhou-Da CAI ; Zhao-Dong HAN ; Yuan-Fa FENG ; Shang-Hua CAI ; Qing-Biao CHEN ; Jian-Guo ZHU ; Wei-De ZHONG
Asian Journal of Andrology 2022;24(5):540-548
The limited treatment options for advanced prostate cancer (PCa) lead to the urgent need to discover new anticancer drugs. Mannose, an isomer of glucose, has been reported to have an anticancer effect on various tumors. However, the anticancer effect of mannose in PCa remains unclear. In this study, we demonstrated that mannose inhibits the proliferation and promotes the apoptosis of PCa cells in vitro, and mannose was observed to have an anticancer effect in mice without harming their health. Accumulation of intracellular mannose simultaneously decreased the mitochondrial membrane potential, increased mitochondrial and cellular reactive oxygen species (ROS) levels, and reduced adenosine triphosphate (ATP) production in PCa cells. Mannose treatment of PCa cells induced changes in mitochondrial morphology, caused dysregulated expression of the fission protein, such as fission, mitochondrial 1 (FIS1), and enhanced the expression of proapoptotic factors, such as BCL2-associated X (Bax) and BCL2-antagonist/killer 1 (Bak). Furthermore, lower expression of mannose phosphate isomerase (MPI), the key enzyme in mannose metabolism, indicated poorer prognosis in PCa patients, and downregulation of MPI expression in PCa cells enhanced the anticancer effect of mannose. This study reveals the anticancer effect of mannose in PCa and its clinical significance in PCa patients.
Animals
;
Apoptosis
;
Cell Line, Tumor
;
Humans
;
Male
;
Mannose
;
Membrane Potential, Mitochondrial
;
Mice
;
Mitochondria
;
Prostatic Neoplasms
;
Reactive Oxygen Species
4.Serum levels of degraded monosaccharides in children with Henoch-Schönlein purpura.
Zhi-Liang WANG ; Kai-Di SHENG ; Yi LIN ; Qiu-Ye ZHANG ; Li-Juan ZHANG ; Hong CHANG
Chinese Journal of Contemporary Pediatrics 2022;24(8):894-898
OBJECTIVES:
To examine the serum levels of degraded monosaccharides in children with Henoch-Schönlein purpura (HSP) and to study the clinical significance of degraded monosaccharides in HSP.
METHODS:
A prospective analysis was performed on 132 children who were diagnosed with HSP from September 2019 to January 2022, and 132 healthy children were enrolled as the control group. High-performance liquid chromatography was used to determine the content of degraded monosaccharides in serum in both groups. The receiver operating characteristic (ROC) curve was used to evaluate the efficiency of degraded monosaccharides for the diagnosis of HSP.
RESULTS:
Compared with the control group, the HSP group had significantly higher serum levels of mannose, glucosamine, aminogalactose, and galactose (P<0.001). The four degraded monosaccharides had an area under the ROC curve of 0.919, 0.913, 0.832, and 0.932 respectively for the diagnosis of HSP (P<0.05).
CONCLUSIONS
Children with HSP have higher serum levels of mannose, glucosamine, aminogalactose, and galactose than the healthy population. The levels of degraded monosaccharides may have an important value for the diagnosis of HSP.
Child
;
Galactose
;
Glucosamine
;
Humans
;
IgA Vasculitis
;
Mannose
;
Monosaccharides
5.Lycium barbarum Polysaccharides Promotes Mitochondrial Biogenesis and Energy Balance in a NAFLD Cell Model.
Yan-Nan ZHANG ; Yi-Qiong GUO ; Yan-Na FAN ; Xiu-Juan TAO ; Qing-Han GAO ; Jian-Jun YANG
Chinese journal of integrative medicine 2022;28(11):975-982
OBJECTIVE:
To explore the protective effect and underlying mechanism of Lycium barbarum polysaccharides (LBP) in a non-alcoholic fatty liver disease (NAFLD) cell model.
METHODS:
Normal human hepatocyte LO2 cells were treated with 1 mmol/L free fatty acids (FFA) mixture for 24 h to induce NAFLD cell model. Cells were divided into 5 groups, including control, model, low-, medium- and high dose LBP (30,100 and 300 µg/mL) groups. The monosaccharide components of LBP were analyzed with high performance liquid chromatography. Effects of LBP on cell viability and intracellular lipid accumulation were assessed by cell counting Kit-8 assay and oil red O staining, respectively. Triglyceride (TG), alanine aminotransferase (ALT), aspartate aminotransferase (AST), adenosine triphosphate (ATP) and oxidative stress indicators were evaluated. Energy balance and mitochondrial biogenesis related mRNA and proteins were determined by quantitative real-time polymerase chain reaction and Western blot, respectively.
RESULTS:
Heteropolysaccharides with mannose and glucose are the main components of LBP. LBP treatment significantly decreased intracellular lipid accumulation as well as TG, ALT, AST and malondialdehyde levels (P<0.05 or P<0.01), increased the levels of superoxide dismutase, phospholipid hydroperoxide glutathione peroxidase, catalase, and ATP in NAFLD cell model (P<0.05). Meanwhile, the expression of uncoupling protein 2 was down-regulated and peroxisome proliferator-activated receptor gamma coactivator-1α/nuclear respiratory factor 1/mitochondrial transcription factor A pathway was up-regulated (P<0.05).
CONCLUSION
LBP promotes mitochondrial biogenesis and improves energy balance in NAFLD cell model.
Humans
;
Non-alcoholic Fatty Liver Disease/drug therapy*
;
Lycium/metabolism*
;
Catalase/metabolism*
;
Organelle Biogenesis
;
Alanine Transaminase
;
Uncoupling Protein 2
;
Fatty Acids, Nonesterified
;
Mannose
;
Nuclear Respiratory Factor 1/metabolism*
;
PPAR gamma/metabolism*
;
Phospholipid Hydroperoxide Glutathione Peroxidase
;
Drugs, Chinese Herbal/pharmacology*
;
Malondialdehyde/metabolism*
;
Superoxide Dismutase/metabolism*
;
Polysaccharides/pharmacology*
;
Triglycerides
;
RNA, Messenger
;
Aspartate Aminotransferases
;
Glucose
;
Adenosine Triphosphate
6.Roles of pattern recognition receptors in diabetic nephropathy.
Zhi-Feng ZHOU ; Lei JIANG ; Qing ZHAO ; Yu WANG ; Jing ZHOU ; Qin-Kai CHEN ; Jin-Lei LV
Journal of Zhejiang University. Science. B 2020;21(3):192-203
Diabetic nephropathy (DN) is currently the most common complication of diabetes. It is considered to be one of the leading causes of end-stage renal disease (ESRD) and affects many diabetic patients. The pathogenesis of DN is extremely complex and has not yet been clarified; however, in recent years, increasing evidence has shown the important role of innate immunity in DN pathogenesis. Pattern recognition receptors (PRRs) are important components of the innate immune system and have a significant impact on the occurrence and development of DN. In this review, we classify PRRs into secretory, endocytic, and signal transduction PRRs according to the relationship between the PRRs and subcellular compartments. PRRs can recognize related pathogen-associated molecular patterns (PAMPs) and danger-associated molecular patterns (DAMPs), thus triggering a series of inflammatory responses, promoting renal fibrosis, and finally causing renal impairment. In this review, we describe the proposed role of each type of PRRs in the development and progression of DN.
Alarmins/physiology*
;
C-Reactive Protein/physiology*
;
Diabetic Nephropathies/etiology*
;
Endocytosis
;
Humans
;
Immunity, Innate
;
Mannose-Binding Lectin/physiology*
;
Pathogen-Associated Molecular Pattern Molecules
;
Receptors, Pattern Recognition/physiology*
;
Serum Amyloid P-Component/physiology*
;
Signal Transduction
7.Study of mycelial polysaccharide from Paraisaria dubia of Ophiocordyceps gracilis asexual.
Yue WANG ; Zhao-Lan LI ; Fei-Ya SUO ; Dong-Ping SUN
China Journal of Chinese Materia Medica 2019;44(8):1704-1709
In order to provide a foundation for the development and application of Ophiocordyceps gracilis and increase the new resources of cordyceps,an asexual Paraisaria dubia was isolated from an O. gracilis fruit body. After 10 days of liquid fermentation,white globular mycelium and clear transparent fermentation were produced. The mycelium was extracted by hot water and precipitated with ethanol to obtain intracellular crude polysaccharide. The protein was deproteinized to obtain deproteinized polysaccharide. The intracellular pure polysaccharide was purified by Sepharose 4 B column chromatography and were analyzed by UV,IR,1 H-NMR,and13 CNMR data,as well as GC and HPLC. The results showed that the intracellular polysaccharide of P. dubia was composed of glucose,galactose and mannose with a molar ratio of 25. 54 ∶2 ∶1. It was a β-configuration glycosylic bond,containing pyranoside. The initial connection of polysaccharide was β(1→2)(1→4)(1→6) connection. This experiment provides a theoretical basis for the development and application of P. dubia.
Fungal Polysaccharides
;
chemistry
;
Galactose
;
Glucose
;
Hypocreales
;
chemistry
;
Mannose
;
Mycelium
;
chemistry
8.Serial Changes in Mannose-Binding Lectin in Patients with Sepsis.
Jin Won HUH ; Kyuyoung SONG ; Hwa Jung KIM ; Jung Sun YUM ; Sang Bum HONG ; Chae Man LIM ; Younsuck KOH
Tuberculosis and Respiratory Diseases 2018;81(4):305-310
BACKGROUND: Mannose-binding lectin (MBL) deficiency leads to increased susceptibility to infection. We investigated whether serial changes in MBL levels are associated with the prognosis of patients diagnosed with septic shock, and correlated with cytokine levels. METHODS: We enrolled 131 patients with septic shock in the study. We analyzed the serum samples for MBL and cytokine levels at baseline and 7 days later. Samples on day 7 were available in 73 patients. RESULTS: We divided the patients with septic shock into four groups according to serum MBL levels ( < 1.3 µg/mL or ≥1.3 µg/mL) on days 1 and 7. Patients with low MBL levels on day 1 and high MBL levels on day 7 showed a favorable prognosis for 28-day survival (odds ratio, 1.96, 95% confidence interval, 1.10–2.87; p=0.087). The high MBL group on day 7 showed a significant decrease in monocyte chemoattractant protein 1, interleukin (IL)-1β, IL-6, IL-8, interferon-γ, and granulocyte macrophage colony-stimulating factor levels compared with the low MBL group on day 7. CONCLUSION: The increase in MBL levels of patients with septic shock may suggest a favorable prognosis and attenuate pro-inflammatory and anti-inflammatory responses.
Chemokine CCL2
;
Cytokines
;
Granulocytes
;
Humans
;
Interleukin-6
;
Interleukin-8
;
Interleukins
;
Macrophage Colony-Stimulating Factor
;
Mannose-Binding Lectin*
;
Prognosis
;
Sepsis*
;
Shock, Septic
9.Analysis of Gene Expression in Human Dermal Fibroblasts Treated with Senescence-Modulating COX Inhibitors.
Genomics & Informatics 2017;15(2):56-64
We have previously reported that NS-398, a cyclooxygenase-2 (COX-2)–selective inhibitor, inhibited replicative cellular senescence in human dermal fibroblasts and skin aging in hairless mice. In contrast, celecoxib, another COX-2–selective inhibitor, and aspirin, a non-selective COX inhibitor, accelerated the senescence and aging. To figure out causal factors for the senescence-modulating effect of the inhibitors, we here performed cDNA microarray experiment and subsequent Gene Set Enrichment Analysis. The data showed that several senescence-related gene sets were regulated by the inhibitor treatment. NS-398 up-regulated gene sets involved in the tumor necrosis factor β receptor pathway and the fructose and mannose metabolism, whereas it down-regulated a gene set involved in protein secretion. Celecoxib up-regulated gene sets involved in G2M checkpoint and E2F targets. Aspirin up-regulated the gene set involved in protein secretion, and down-regulated gene sets involved in RNA transcription. These results suggest that COX inhibitors modulate cellular senescence by different mechanisms and will provide useful information to understand senescence-modulating mechanisms of COX inhibitors.
Aging
;
Animals
;
Aspirin
;
Celecoxib
;
Cell Aging
;
Cyclooxygenase 2
;
Fibroblasts*
;
Fructose
;
Gene Expression*
;
Genes, vif
;
Humans*
;
Mannose
;
Metabolism
;
Mice
;
Mice, Hairless
;
Oligonucleotide Array Sequence Analysis
;
RNA
;
Skin Aging
;
Tumor Necrosis Factor-alpha
10.Association of MBL With Work-Related Respiratory Symptoms in Bakery Workers.
Mi Ae KIM ; Moon Kyung YOON ; Seung Hyun KIM ; Hae Sim PARK
Allergy, Asthma & Immunology Research 2017;9(1):85-91
Baker's asthma is the most prevalent occupational asthma, and IgE-mediated response is known as a major pathogenesis. However, recent studies have suggested the involvement of innate immune response because wheat flour contains bacterial endotoxins or lipopolysaccharides. To further understand a role of innate immune response in the development of work-related respiratory symptoms (WRS) in bakery workers, we investigated mannose-binding lectin (MBL), one of the initiating components of the complement cascade in a single cohort of bakery workers. A total of 373 bakery workers completed a questionnaire regarding WRS. The bakery workers were divided into 2 groups according to previous history of allergic rhinitis (AR)/bronchial asthma (BA): those with history of AR/BA (group I) and those without (group II). We measured serum MBL levels by using enzyme-linked immunosorbant assay and genotyped 4 single nucleotide polymorphisms of the MBL2 gene (226G>A in exon 1, -554G>C, -431A>C, and -225G>C in the promoter) by using TaqMan assays. Fifty-nine subjects (15.5%) were previously diagnosed with AR/BA, and 64 subjects (16.8%) complained of WRS. No significant differences were found in serum MBL levels between groups I and II. However, in group II subjects, but not in group I subjects, the serum MBL levels were significantly higher in bakery workers with WRS than in those without. In addition, the serum MBL levels were significantly different according to genetic polymorphisms of the MBL2 gene and its haplotypes. In conclusion, serum MBL, affected by genetic polymorphisms, may be associated with WRS in bakery workers with no previous history of AR/BA.
Asthma
;
Asthma, Occupational
;
Cohort Studies
;
Complement System Proteins
;
Endotoxins
;
Exons
;
Flour
;
Haplotypes
;
Immunity, Innate
;
Lipopolysaccharides
;
Mannose-Binding Lectin
;
Polymorphism, Genetic
;
Polymorphism, Single Nucleotide
;
Rhinitis, Allergic
;
Triticum

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