1.Improvement of neutral protease activity of Bacillus amyloliquefaciens LX-6 by combined ribosome engineering and medium optimization and its application in soybean meal fermentation.
Yifan ZHU ; Xinyi HUANG ; Tao HAN ; Jiteng WANG ; Xiaoping YU ; Zheng MA
Journal of Zhejiang University. Science. B 2025;26(8):805-812
Soybean meal (SBM) prepared by soybean crushing is the most popular protein source in the poultry and livestock industries (Cai et al., 2015) due to its economic manufacture, high protein content, and good nutritional value. Despite these benefits, SBM contains various antigen proteins such as glycinin and β-conglycinin, which account for approximately 70% of the total proteins of the SBM and reduce digestibility and damage intestinal function (Peng et al., 2018). Treating SBM with proteases (neutrase, alcalase, and trypsin) or fermentation can eliminate these antigen proteins (Contesini et al., 2018). Because of its safety and rapid growth cycle, Bacillus strains are considered ideal for the fermentation industry (Yao et al., 2021). SBM fermented by Bacillus yields products with high nutritional value and low levels of antinutritional factors (ANFs), stimulating research in this area (Yuan et al., 2017). Kumari et al. (2023) demonstrated that fermentation with Bacillus species effectively degrades antigen proteins and increases crude protein content. The degradation of antigen proteins relies on protease hydrolysis. Low protease production is the major obstacle hindering the widespread use of microbial fermentation techniques.
Bacillus amyloliquefaciens/metabolism*
;
Fermentation
;
Glycine max/metabolism*
;
Soybean Proteins/metabolism*
;
Peptide Hydrolases/metabolism*
;
Ribosomes/metabolism*
;
Globulins
;
Antigens, Plant
;
Seed Storage Proteins
2.Effect of Astragalus mongholicus polysaccharides on gene expression profiles of dendritic cells isolated from healthy donors.
Chaojun CHEN ; Qiang FU ; Yuejun LI ; Zhiliang LI
Journal of Southern Medical University 2015;35(12):1802-1805
OBJECTIVETo investigate the anti-atherosclerosis mechanism of Astragalus mongholicus polysaccharides (APS) by examining its effect on gene expression profiles of the dendritic cells (DCs) from healthy donors.
METHODSPeripheral blood DCs from healthy donors were incubated with 200 mg/L APS overnight, and changes in the gene expression profiles were investigated using microarray technique and RT-PCR.
RESULTSCompared with the control cells, APS-treated DCs showed significantly up-regulated expressions of CD36 (0.97 ± 0.23 vs 5.45 ± 1.14) and IL-27 (1.08 ± 0.22 vs 2.97 ± 0.61) and down-regulated expression of expression of IFI16 (0.98 ± 0.18 vs 0.46 ± 0.11).
CONCLUSIONSAPS can promote the maturation and differentiation of DCs by up-regulating CD36 and IL-27 and down-regulating IFI16, and thus positively affects the occurrence and progression of the atherosclerosis.
Astragalus Plant ; chemistry ; CD36 Antigens ; metabolism ; Cell Differentiation ; Dendritic Cells ; drug effects ; Humans ; Interleukins ; metabolism ; Nuclear Proteins ; metabolism ; Phosphoproteins ; metabolism ; Polysaccharides ; pharmacology ; Transcriptome
3.In vivo digestive stability of soybean β-conglycinin β-subunit in WZS minipigs.
Qiong HUANG ; Hai Bin XU ; Zhou YU ; Shan LIU ; Peng GAO
Biomedical and Environmental Sciences 2015;28(1):85-88
By now, the digestive stability experiments provided by most authoritative organizations are in vitro tests. Evaluating the protein digestive stability with in vivo models should be more objective. The present study aimed to verify the in vivo digestibility of soybean β-conglycinin β-subunit in Wuzhishan (WZS) minipigs. Three minipigs were surgically fitted with O-stomach and T-ileum cannulae and fed with soybean meals. According to SDS-PAGE, the 50 kD fraction of soybean β-conglycinin β-subunit persisted in the gastric fluid until 6 h after feeding, which was detected at 3 h and clearly visible at 4-6 h in the intestinal fluid. Western blot with anti-β-conglycinin β-subunit McAb confirmed it.
Animals
;
Antigens, Plant
;
chemistry
;
metabolism
;
Digestion
;
physiology
;
Globulins
;
chemistry
;
metabolism
;
Male
;
Protein Subunits
;
chemistry
;
metabolism
;
Seed Storage Proteins
;
chemistry
;
metabolism
;
Soybean Proteins
;
chemistry
;
metabolism
;
Swine
;
Swine, Miniature
;
physiology
4.Preparation and identification of artificial antigen for rhein.
Bo ZHANG ; Yuan YUAN ; Lu-qi HUANG ; Bao-min WANG ; Tie-gui NAN
China Journal of Chinese Materia Medica 2015;40(8):1463-1467
The Rhei Radix et Rhizoma was one of the most widely used traditional Chinese medicine for its special biological activities. The content of rhein, one of its major compounds, was an important standard for the quantity control of Rhei Radix et Rhizoma. The major method used for the detection of rhein was instrumental analysis like HPLC, but it was complex, time-consuming and cannot detect large samples at the same time. The enzyme-linked imunmosorbent assay (ELISA) was accurate, reliable, simple, low costs, and of a high-throughout. Recently, it was widely used for the determination of those small molecule compounds in some traditional Chinese medicinal plants. In this study, an artificial antigen were synthesized by the carbodiimide (CDI) method. Rhein-bovine (rhein-BSA) conju gate and rhein-ovalbumin (rhein-OVA) conjugate, were produced as the immunogen and coating antigen, respectively. The conjugate and the hapten number in the conjugate were determined by UV-Vis spectrophotometry (UV). The conjugation ratio of Rhein and BSA was about 4.0:1, rhein acid and OVA was 2.6 : 1, respectively. Rhein-BSA conjugate was used to immunize Bal b/c mice to produce antiserum. The antiserum titer of the Rhein were higher than 8000 detected by ELISA. The successfully synthesized conjugate antigen rhein-BSA implies its feasibility in the establishment of fast immunoassay for the rhein content determination.
Animals
;
Anthraquinones
;
analysis
;
immunology
;
Antibodies
;
analysis
;
immunology
;
Antigens, Plant
;
analysis
;
immunology
;
Drugs, Chinese Herbal
;
analysis
;
Enzyme-Linked Immunosorbent Assay
;
instrumentation
;
methods
;
Mice
;
Mice, Inbred BALB C
;
Rheum
;
chemistry
;
immunology
;
Rhizome
;
chemistry
;
immunology
5.Agrobacterium-mediated transformation of LJAMP2 gene into 'Red Sun' kiwifruit and its molecular identification.
Yue ZHOU ; Xupeng ZHAO ; Xiuhua WU ; Yanling ZHANG ; Lin ZHANG ; Keming LUO ; Shaohu TANG
Chinese Journal of Biotechnology 2014;30(6):931-942
Bacterial canker caused by Pseudomonas syringae pv. Actinidiae is one of the most important diseases of kiwifruit (Actinidia chinensis) and leads to considerable yield losses. In order to obtain transgenic plants with resistance for 'Red Sun' kiwifruit to canker disease, a non-specific lipid transfer protein-like antimicrobial protein gene (LJAMP2) from motherwort (Leonurus japonicus) was introduced into 'Red Sun' kiwifruit through Agrobacterium-mediated transformation. After two days of co-cultivation with A. tumefaciens strain LBA4404 harboring 35S:LJAMP2, the transformed explants were transferred to the selection medium containing 25 mg/L kanamycin+3.0 mg/L BA+1.0 mg/L NAA. The regeneration efficiency of kanamycin-resistant shoots reached to 85%. All (100%) of kanamycin-resistant shoots rooted on half-strength MS medium supplemented with 0.8 mg/L IBA and a total of 40 regenerated plantlets were obtained. PCR and histochemical GUS activity analysis show that 23 of 40 lines (57.50%) were positive, suggesting that the LJAMP2 gene was integrated into the genome of 'Red Sun' kiwifruit. Taken together, we established an efficient genetic transformation method for 'Red Sun' kiwifruit using A. tumefaciens and the transformation frequency reached 5.11%. This protocol will be useful for the genetic breeding of 'Red Sun' kiwifruit for improvement of disease resistance.
Actinidia
;
genetics
;
Agrobacterium
;
Antigens, Plant
;
genetics
;
Carrier Proteins
;
genetics
;
Leonurus
;
Plant Proteins
;
genetics
;
Plants, Genetically Modified
;
genetics
;
Transformation, Genetic
6.Comparison of Specific IgE Antibodies to Wheat Component Allergens in Two Phenotypes of Wheat Allergy.
Young Hee NAM ; Eui Kyung HWANG ; Hyun Jung JIN ; Jeong Min LEE ; Yoo Seob SHIN ; Young Min YE ; Arantxa PALACIN ; Gabriel SALCEDO ; Soo Young LEE ; Hae Sim PARK
Journal of Korean Medical Science 2013;28(11):1697-1699
Specific IgE to gliadin was proposed as a marker for wheat dependent exercise induced anaphylaxis, while Tri a 14 was found to induce IgE response in baker's asthma. We evaluated whether these components could be used for discriminating phenotypes of wheat allergy. Twenty-nine patients who were wheat-induced anaphylaxis and/or urticaria (n=21, group I) and baker's asthma (n=8, group II) were enrolled. The prevalence of serum specific IgE to Tri a 14 was higher in group II (25%) than in group I (4.8%), while the serum specific IgE to gliadin was significantly higher in group I (70%) than in group II (12.5%). The cutoff value for predicting the baker's asthma using the ratio of serum specific IgE to Tri a 14 to gliadin was 742.8 optical densityx1,000/(kU/L) with high sensitivity and specificity. These findings suggest that Tri a 14/gliadin may be a potential marker for predicting baker's asthma.
Adult
;
Anaphylaxis/immunology
;
Antigens, Plant/*immunology
;
Asthma/blood/diagnosis/immunology
;
Biological Markers/blood
;
Carrier Proteins/*immunology
;
Female
;
Gliadin/*immunology
;
Humans
;
Immunoglobulin E/*blood/immunology
;
Male
;
Phenotype
;
Triticum/immunology
;
Urticaria/immunology
;
Wheat Hypersensitivity/*diagnosis/*immunology
7.Effect of astragalus polysaccharide on sensitivity of leukemic cell line HL-60 to NK cell cytotoxicity and its mechanism.
Peng-Yun ZENG ; Li-Li DENG ; Ling-Ling YUE ; Lian-Sheng ZHANG
Journal of Experimental Hematology 2012;20(4):880-883
The objective of this study was to explore the effect of astragalus polysaccharide (APS) on sensitivity of leukemic cell line HL-60 to NK cell cytotoxicity and its mechanism. The cytotoxicities of NK cells against HL-60 cells were analyzed by LDH releasing assay at different effect-to-target cell ratios (E:T) before and after treated with APS. The gene expression of MHC class I chain-related (MICA) in HL-60 cells before and after APS treatment was assayed with RT-PCR. Protein expression of MICA in HL-60 cells was assayed by flow cytometry before and after treated by APS. The results showed that after treated with APS 15 mg/ml for 48 h, the cytotoxicities of NK cells against HL-60 cells enhanced at different effect-to-target (P < 0.05), and the gene and protein expressions in MICA of HL-60 cells were up-regulated (P < 0.05). It is concluded that the APS can obviously up-regulate the expression of MICA in HL-60 cells, thus enhance sensitivity of HL-60 cells to cytotoxicity of NK cells.
Astragalus Plant
;
Cytotoxicity, Immunologic
;
drug effects
;
HL-60 Cells
;
Histocompatibility Antigens Class I
;
metabolism
;
Humans
;
Killer Cells, Natural
;
Polysaccharides
;
pharmacology
8.Subcellular localization and resistance to Botrytis cinerea of a new type lipid transfer protein AtDhyPRP1 from Arabidopsis thaliana.
Chen ZHANG ; Lan LI ; Ziqin XU
Chinese Journal of Biotechnology 2012;28(5):602-612
Genetic transformation was adopted to analyze the subcellular localization and the resistance to fungal pathogens of Arabidopsis lipid transfer protein AtDHyPRP1. The coding sequence of AtDHyPRP1 amplified by PCR from Ws ecotype was used to construct the plant binary expression vector pRI101-AN-AtDHyPRP1 and the fusion expression vector pCAMBIA1302-AtDHyPRP1-GFP. Transgenic tobacco and Arabidopsis plants were produced by leaf disc and floral dip protocols, respectively. AtDHyPRP1 could improve the resistance of tobacco to Botrytis cinerea remarkably and the infection sites on transgenic tobacco leaves accumulated large amounts of H2O2. Observation under laser scanning confocal microscope showed that AtDHyPRP1 was localized to cell surface. It suggested that AtDHyPRP1 might play special function after secretion to outside of the cell and was involved in plant defense system against pathogens.
Amino Acid Sequence
;
Antigens, Plant
;
genetics
;
metabolism
;
Arabidopsis
;
genetics
;
metabolism
;
microbiology
;
Arabidopsis Proteins
;
genetics
;
metabolism
;
Botrytis
;
Carrier Proteins
;
genetics
;
metabolism
;
Disease Resistance
;
Escherichia coli
;
genetics
;
metabolism
;
Molecular Sequence Data
;
Plant Diseases
;
immunology
;
microbiology
;
Plant Proteins
;
genetics
;
metabolism
;
Plants, Genetically Modified
;
genetics
;
metabolism
;
microbiology
;
Recombinant Proteins
;
genetics
;
metabolism
;
Subcellular Fractions
;
metabolism
;
Tobacco
;
genetics
;
metabolism
;
microbiology
9.Effects of Radix Astragali on IL-1beta, TNF-alpha and antigen expression of peripheral blood mononuclear cells in patients with Graves disease.
Juan WU ; Dong-fang LIU ; Yu CHEN
Chinese Journal of Integrated Traditional and Western Medicine 2011;31(11):1487-1490
OBJECTIVETo study the effects of Radix Astragali on serum cytokines IL-1beta, TNFalpha and antigen expression of peripheral blood mononuclear cells (PBMCs) in patients with Graves disease (GD).
METHODSEighty GD patients at their first visit were randomly assigned to the methimazole (MMI) group (Group A) and the MMI combined Radix Astragali group (Group B), 40 in each. The improvement of clinical symptoms and thyroid functions were observed after one-month treatment. The serum IL-1beta and TNF-alpha levels in the peripheral blood were determined using radioimmunoassay. The expression levels of surface antigen CD80, CD54, and HLA-DR of PBMCs were detected using flow cytometry.
RESULTSThe improvement of the thyroid gland function was similar in the two groups. There was no obvious change in the levels of autoantibody TGAb or TPOAb of the two groups. Symptoms such as fear of heat, hidrosis, palpitation, and so on were more obviously improved in Group B than in Group A (P < 0.05). The serum IL-betaP, TNFalphaa, CD00 levels in the peripheral blood were all improved in the two groups after treatment when compared with before treatment ( P < 0.05 or P < 0.01). But the serum levels of IL-beta and TNFalpha decreased more obviously in Group B than in Group A ( P < 0.05). The expression of CD54 decreased more obviously in Group B (P < 0.01), showing statistical difference when compared with Group A at the same time point (P < 0.05).
CONCLUSIONRadix Astragali could significantly relieve the clinical symptoms such as hidrosis and palpitation, regulate the immune function of GD patients, playing an important role in the adjuvant therapy for GD.
Adult ; Astragalus Plant ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Graves Disease ; blood ; drug therapy ; immunology ; HLA Antigens ; metabolism ; Humans ; Interleukin-1beta ; blood ; Leukocytes, Mononuclear ; drug effects ; immunology ; metabolism ; Male ; Methimazole ; therapeutic use ; Middle Aged ; Tumor Necrosis Factor-alpha ; blood
10.Progress of researches on the allergens Ara h 1, Ara h 2 and Ara h 3 from peanut.
Shengjuan JIANG ; Zhengyi ZHOU ; Yujun SUN ; Songhua WANG
Journal of Biomedical Engineering 2010;27(6):1401-1405
Peanut is one of the most popular foods in the world due to its high nutrition; however, it contains multiple seed storage proteins which are identified as allergens and hence are the most common cause of life-threatening, IgE-mediated anaphylaxis among the hypersensitive individuals. Three peanut proteins, Arachis hypogaea allergy 1, 2, 3 (Ara h 1, Ara h 2 and Ara h 3), which have the common biochemical characteristics like resistance to proteases and heat, are considered as the major allergens because they are recognized by serum IgE from a peanut-allergic patient population. The linear IgE-binding epitopes in the allergens lay the foundation of the anaphylaxis in the peanut-allergic individuals. Peanut allergy is often a life-long problem, so many investigators are focusing on decreasing clinical reactivity. In this review, the latest advances in the researches on biochemical characteristics, structure and function of the three major allergens were described and particular attention was given to the immunity properties of the three allergens. The future research directions were also discussed.
2S Albumins, Plant
;
chemistry
;
genetics
;
immunology
;
Animals
;
Antigens, Plant
;
chemistry
;
genetics
;
immunology
;
Arachis
;
chemistry
;
DNA
;
genetics
;
Glycoproteins
;
chemistry
;
genetics
;
immunology
;
Humans
;
Immunoglobulin E
;
genetics
;
Plant Proteins
;
chemistry
;
genetics
;
immunology

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