1.Research progress of heparinase in the field of medicine.
Wenli LIU ; Yingzi JIANG ; Liqing ZHAO ; Peixin ZHANG ; Shulan WANG
Chinese Journal of Biotechnology 2018;34(12):1953-1962
Heparinases can produce biologically active oligosaccharides by specifically cleaving the α-(1,4) glycosidic linkages of heparin and heparan sulphate. Heparinases are divided into heparinase and heparanase. Because heparinase is an effective biocatalyst, more and more researchers pay attention to the application of heparinase in medical field in the recent years. Combined with the related research work in our group, the application value of heparinase in the medical field was summarized, such as the determination of the structure of heparin, the preparation of low-molecular-weight heparin and ultra-low-molecular-weight heparin, tumor therapy and as a heparin antagonist. In addition, we summarized the definition, source of heparinase and its application in the medicine field. Heparinases have a great application prospect in the field of medicine.
Heparin
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Heparin Lyase
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metabolism
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Heparitin Sulfate
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Oligosaccharides
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Polysaccharide-Lyases
2.Effects of the dietary supplementation with fructooligosaccharides on the excretion of nitrogen and phosphorus in Miichthys miiuy fries.
Tian-xing WU ; Zeng-fu SONG ; Li-sheng CAI ; Xue-yan DING ; Qing-sen YU
Journal of Zhejiang University. Science. B 2005;6(8):798-802
Effects of dietary supplementation with fructooligosaccharides on the excretion of nitrogen and phosphorus in Miichthys miiuy fries were investigated. Nine hundred Miichthys miiuy fries were divided into 3 groups, each with triplicates. The basal diet and the basal diet supplemented with carnitine groups were considered as the negative and positive controls respectively. Results showed that the nitrogen concentration in excreted feces decreased significantly in fries fed the diet supplementation with 1000 x 10(-6) fructooligosaccharides and 200 x 10(-6) carnitine (P<0.05). The ammonic-nitrogen concentration decreased significantly in the carnitine group only (P<0.05), indicating the decreasing tendency caused by the supplementation with fructooligosaccharides. Supplementation with both did not have significant effects on the concentration of phosphorus in feces of Miichthys miiuy fries.
Administration, Oral
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Animals
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Dietary Supplements
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Feces
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Fishes
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metabolism
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Nitrogen
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metabolism
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Oligosaccharides
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administration & dosage
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Phosphorus
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metabolism
3.Effect of continuous temperature change on hydrolytic products of yeast beta-glucan by endo-beta-1,3-glucanase.
Feng DUAN ; Xuemei LU ; Yongcheng DUAN ; Peiji GAO
Chinese Journal of Biotechnology 2011;27(7):1092-1099
In order to explore the influence of reaction temperature on the product composition, the effect of continuous temperature change (22 degrees C-60 degrees C, +/-0.1 degree C) on hydrolysis of yeast beta-glucan by endo-beta-1,3-glucanase was determined by using self-developed Biochem-temperature Characteristic Apparatus. The activation energy of enzymatic hydrolysis of yeast beta-glucan was 84.17 kJ/mol. The optimum temperature represented by accumulation of products decreased exponentially within a certain period of time. The components of the products were changed with reaction temperature. The length of oligosaccharides decreased with the increase of temperature. The main products were laminaribiose and laminaritriose at the temperature higher than 46 degrees C, while the main products were laminaripentaose and larger molecular weight components at the temperature lower than 30 degrees C. The results can provide precise parameters to control the reaction temperature of the production of 1,3-beta-D-glucooligosaccharides.
Enzyme Activation
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Glucan Endo-1,3-beta-D-Glucosidase
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chemistry
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metabolism
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Hydrolysis
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Oligosaccharides
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chemistry
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metabolism
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Temperature
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Yeasts
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metabolism
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beta-Glucans
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metabolism
4.Construction of engineered Streptococcus zooepidemicus for the production of hyaluronic acid ligosaccharide.
Chaobao WEI ; Guocheng DU ; Jian CHEN ; Zhen KANG
Chinese Journal of Biotechnology 2019;35(5):805-815
Hyaluronic acid (HA) is widely used in many fields, such as medicine, cosmetics and food. The bioactivity of HA depends on its molecular weight (Mw). Owing to the important physiological activities and special physiological functions, HA oligosaccharides have important application prospects in medicine fields. Streptococcus zooepidemicus has wide applications in commercial production of HA, due to its short fermentation cycle and strong production intensity. In order to efficiently synthesize HA oligosaccharides and solve the dissolved oxygen in the fermentation process, in this study, we overexpressed HA synthase (HasA) and introduced and optimized the leech hyaluronidase LHAase in Streptococcus zooepidemicus WSH-24. As a result, HA oligosaccharides were efficiently produced with improved dissolved oxygen. After 24 h, HA oligosaccharides production intensity reached to 294.2 mg/(L·h), and the concentration accumulated to 0.97 g/L in flask cultures, which was 1.82 times of the wild strain. Impressively, HA oligosaccharides were increased to 7.06 g/L in 3 L fermentor. The constructed Streptococcus zooepidemicus strain for producing HA oligosaccharides would have broad application prospects.
Bioreactors
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Fermentation
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Hyaluronan Synthases
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genetics
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metabolism
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Hyaluronic Acid
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genetics
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metabolism
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Industrial Microbiology
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Oligosaccharides
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genetics
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metabolism
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Streptococcus equi
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genetics
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metabolism
5.Expression of SleX in nasal epithelium with chronic rhinosinusitis.
Xuewei ZHU ; Dongdong ZHU ; Kewei SUN ; Yusheng WANG ; Zhen DONG
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2008;22(20):917-919
OBJECTIVE:
To explore the expression of SleX in nasal epithelium in patients with chronic rhinosinusitis.
METHOD:
Nine nasal epitheliums from patients with chronic rhinosinusitis and 7 normal controls were stained with SleX antibody by immunohistochemistry, and its expression were analyzed.
RESULT:
Both nasal epithelial cells and neutrophils expressed SleX. Nearly 88.9% nasal epithelium from patients with chronic rhinosinusitis expressed SleX while 14.3% in normal control. Expression of SleX in nasal epitheliums from patients with chronic rhinosinusitis were higher than that in normal control. There were significant difference between two groups.
CONCLUSION
Nasal epithelium from patients with CRS is highly expressed SleX. It may involve the occurrence of chronic rhinosinusitis.
Adult
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Chronic Disease
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Female
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Humans
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Male
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Middle Aged
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Nasal Mucosa
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metabolism
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Neutrophils
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metabolism
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Oligosaccharides
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analysis
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Sinusitis
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metabolism
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Young Adult
6.Microencapsulation of immunoglobulin Y: optimization with response surface morphology and controlled release during simulated gastrointestinal digestion.
Jin ZHANG ; Huan-Huan LI ; Yi-Fan CHEN ; Li-Hong CHEN ; Hong-Gang TANG ; Fan-Bin KONG ; Yun-Xin YAO ; Xu-Ming LIU ; Qian LAN ; Xiao-Fan YU
Journal of Zhejiang University. Science. B 2020;21(8):611-627
Immunoglobulin Y (IgY) is an effective orally administered antibody used to protect against various intestinal pathogens, but which cannot tolerate the acidic gastric environment. In this study, IgY was microencapsulated by alginate (ALG) and coated with chitooligosaccharide (COS). A response surface methodology was used to optimize the formulation, and a simulated gastrointestinal (GI) digestion (SGID) system to evaluate the controlled release of microencapsulated IgY. The microcapsule formulation was optimized as an ALG concentration of 1.56% (15.6 g/L), COS level of 0.61% (6.1 g/L), and IgY/ALG ratio of 62.44% (mass ratio). The microcapsules prepared following this formulation had an encapsulation efficiency of 65.19%, a loading capacity of 33.75%, and an average particle size of 588.75 μm. Under this optimum formulation, the coating of COS provided a less porous and more continuous microstructure by filling the cracks on the surface, and thus the GI release rate of encapsulated IgY was significantly reduced. The release of encapsulated IgY during simulated gastric and intestinal digestion well fitted the zero-order and first-order kinetics functions, respectively. The microcapsule also allowed the IgY to retain 84.37% immune-activity after 4 h simulated GI digestion, significantly higher than that for unprotected IgY (5.33%). This approach could provide an efficient way to preserve IgY and improve its performance in the GI tract.
Alginic Acid/chemistry*
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Chitin/chemistry*
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Chitosan
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Delayed-Action Preparations
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Digestion
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Drug Compounding
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Drug Liberation
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Gastrointestinal Tract/metabolism*
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Immunoglobulins/metabolism*
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Oligosaccharides
7.Advances in algae tool enzymes: alginate lyases.
Liyan LI ; Huashi GUAN ; Xiaolu JIANG ; Jianjun HAO
Chinese Journal of Biotechnology 2011;27(6):838-845
Marine can be considered as a rather unexplored source of biological material. Production of algal oligosaccharides by using valuable enzymes from marine origin has become an important way to utilize marine resources. As one of algal tool enzymes, the use of alginate lyases has been focused mainly on development and application of alginate oligosaccharides with bioactive function in recent years. In this paper, we reviewed the research of alginate lyases over the past decade in several aspects, including their origin, diversity, substrate specification, mode of action, structure and catalysis mechanism, assay of enzyme activity, enzyme characterization, as well as our own experience on this subject. At the end of the review, the application prospects of alginate lyases are presented.
Alginates
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metabolism
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Glucuronic Acid
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metabolism
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Hexuronic Acids
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metabolism
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Marine Biology
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methods
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Oligosaccharides
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metabolism
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Phaeophyta
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enzymology
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Polysaccharide-Lyases
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classification
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isolation & purification
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metabolism
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Substrate Specificity
8.A novel strain ZG0656 producing alpha-amylase inhibitor and fermentation, separation, properties, and application of its products.
Peng GENG ; Qian SHI ; Qi ZHANG ; Gang BAI
Chinese Journal of Biotechnology 2008;24(6):1103-1107
A strain ZG0656 producing a-amylase inhibitor was isolated from soil in this study. Polyphasic taxonomic studies were performed, including appearance characteristics, culture characteristics, phenotypic characteristics, cell walls chemical composition, nearly complete 16S rDNA sequence alignment with those of representative Streptomyces species. These results revealed that strain ZG0656 represents a novel variation of Streptomyces coelicoflavus, for which we propose the name S. coelicoflavus var. nankaiensis. After fermentation in a 10 L fermentor, alpha-amylase inhibitors were accumulated in the harvested broth of strain ZG0656. The alpha-amylase inhibitors we obtained were identified as aminooligosaccharides after concentration, resin-adsorption, gel-filtration, and desiccation. They could intensively inhibit alpha-amylase, depress postprandial blood glucose elevation obviously. Thus, the a-amylase inhibitors are expected to act as drugs or functional food against diabetes.
Amino Sugars
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biosynthesis
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isolation & purification
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Fermentation
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Oligosaccharides
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biosynthesis
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isolation & purification
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Soil Microbiology
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Streptomyces
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classification
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metabolism
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alpha-Amylases
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antagonists & inhibitors
9.Synthesis and characterization of alginate oligosaccharides produced by Pseudomonas mendocina NK-01.
Wenbin GUO ; Shufang WANG ; Mingfeng CAO ; Weitao GENG ; Cunjiang SONG
Chinese Journal of Biotechnology 2009;25(9):1366-1370
In this study, we found that when Pseudomonas mendocina NK-01 accumulated intracellular carbon reserve, medium chain length poly (3-hydroxyalkanoates), it also synthesized extracellular saccharides, alginate oligosaccharides. The high carbon nitrogen ratio of culture medium facilitated alginate oligosaccharides production. We analyzed the structure of alginate oligosaccharide by Ultraviolet-Visible Spectrophotometry, Fourier Transform Infrared Spectroscopy, 1H and 13C of Nuclear Magnetic Resonance, and found that it was compounded in line from beta-D-mannuronic acids and alpha-L-gluronic acids via beta-(1-->4)/ alpha-(1-->4) bonds, which acetylated partly on the 2- and/or 3-hydroxy. In addition, we determined the weight-average molecular weight of alginate oligosaccharides by gel permeation chromatography to be 2054.
Alginates
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chemistry
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Glucuronic Acid
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biosynthesis
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chemistry
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genetics
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Hexuronic Acids
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chemistry
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Molecular Weight
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Oligosaccharides
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biosynthesis
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chemistry
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genetics
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Pseudomonas mendocina
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metabolism
10.Production and accumulation of xylooligosaccharides with long chains by growing culture and xylanase of a mutant strain of Bacillus pumilus X-6-19.
Qingzhu YUAN ; Tsuyoshi ADACHI ; Shinji TAKENAKA ; Shuichiro MURAKAMI ; Machiko TANAKA ; Kenji AOKI
Chinese Journal of Biotechnology 2008;24(7):1221-1227
Bacillus pumilus X-6-9 isolated from soil and subsequently identified, produced xylooligosaccharides with long chains from xylan and accumulated them in the culture. By improving the culture conditions and mutating the bacterium, a 3.2-fold increase in the production of the xylooligosaccharides was established, when compared to the original culture conditions of B. pumilus X-6-19. The addition of D-glucose to the culture of the mutant strain U-3 of B. pumilus X-6-9 repressed the synthesis of beta-xylosidase, but not xylanase. Thus, it was revealed that strain U-3 was a good organism for the production and accumulation of xylooligosaccharides with long chains from xylan by a microbial culture. Xylanase produced by strain U-3 was purified to homogeneity and characterized. The hydrolyzates generated by the purified xylanase contained xylobiose, xylotriose, xylotetraose, and xylopentaose, but not xylose.
Bacillus
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genetics
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metabolism
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Culture Techniques
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methods
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Endo-1,4-beta Xylanases
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biosynthesis
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genetics
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metabolism
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Glucose
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pharmacology
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Mutation
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Oligosaccharides
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biosynthesis
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chemistry
;
genetics
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Recombinant Proteins
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biosynthesis
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genetics
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metabolism
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Soil Microbiology