1.One step production of isomalto-oligosaccharides by engineered Yarrowia lipolytica yeast co-displayed β-amylase and α-transglucosidase.
Dawen LIU ; Hairong CHENG ; Zixin DENG
Chinese Journal of Biotechnology 2019;35(1):121-132
Isomalto-oligosaccharides (IMO) have good physiochemical properties and excellent physiological functions to make it widely used in food, medicine, feed, cosmetics and other industries. However, the procedures for industrial production of IMO are complicated. Therefore, it is necessary to develop an economical and easy-to-operate method. The genes encoding for β-amylase and α-transglucosidase were fused and co-displayed on the yeast cell surface of Yarrowia lipolytica which can convert liquefied starch to IMO in one step. The highest IMO purity of 75.3% was obtained using the displayed fusion-enzyme at 50 °C. This method showed potential application in IMO production.
Oligosaccharides
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Starch
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Yarrowia
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beta-Amylase
2.Strategies for engineering the thermo-stability of glycosidase.
Rui LIU ; Yu LIU ; Qiaofeng LI ; Xudong FENG ; Chun LI ; Xiaopeng GAO
Chinese Journal of Biotechnology 2021;37(6):1919-1930
Glycosidases are widely used in food and pharmaceutical industries due to its ability to hydrolyze the glycosidic bonds of various sugar-containing compounds including glycosides, oligosaccharides and polysaccharides to generate derivatives with important physiological and pharmacological activity. While glycosidases often need to be used under high temperature to improve reaction efficiency and reduce contamination, most glycosidases are mesophilic enzymes with low activity under industrial production conditions. It is therefore critical to improve the thermo-stability of glycosidases. This review summarizes the recent advances achieved in engineering the thermo-stability of glycosidases using strategies such as directed evolution, rational design and semi-rational design. We also compared the pros and cons of various techniques and discussed the future prospects in this area.
Glycoside Hydrolases/genetics*
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Oligosaccharides
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Polysaccharides
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Protein Engineering
3.Recent advances in the bio-production of human milk oligosaccharides 2'-FL and 3-FL.
Zheng XU ; Na LI ; Yingli CHEN ; Wenrui ZHANG ; Wanying ZHU
Chinese Journal of Biotechnology 2020;36(12):2767-2778
Human milk oligosaccharides (HMO) are important immunoactive components found in breast milk. Scientific research proves that HMOs are significantly beneficial for infant health. 2'-fucosyllactose (2'-FL) is the major component of HMO, which obtained growing attentions from food industry. Besides, 3-fucosyllactose (3-FL) is another important fucosyllactose and it has a similar synthetic route comparing to 2'-FL. Thus, research of the two HMO components has interactive effects for each other. Recently, numerous publications are available for 2'-FL and 3-FL. The microbial cell factory is able to massively produce fucosyllactose via an efficient way, which will show considerable influences in dairy industry. In this paper, we review recent studies on 2'-FL and 3-FL, and discuss their prospects according to published literature and patents.
Female
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Humans
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Infant
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Milk, Human
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Oligosaccharides
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Trisaccharides
4.Chemical synthesis of a synthetically useful L-galactosaminuronic acid building block.
Chun-Jun QIN ; Hong-Li HOU ; Mei-Ru DING ; Yi-Kuan QI ; Guang-Zong TIAN ; Xiao-Peng ZOU ; Jun-Jie FU ; Jing HU ; Jian YIN
Chinese Journal of Natural Medicines (English Ed.) 2022;20(5):387-392
Most bacterial cell surface glycans are structurally unique, and have been considered as ideal target molecules for the developments of detection and diagnosis techniques, as well as vaccines. Chemical synthesis has been a promising approach to prepare well-defined oligosaccharides, facilitating the structure-activity relationship exploration and biomedical applications of bacterial glycans. L-Galactosaminuronic acid is a rare sugar that has been only found in cell surface glycans of gram-negative bacteria. Here, an orthogonally protected L-galactosaminuronic acid building block was designed and chemically synthesized. A synthetic strategy based on glycal addition and TEMPO/BAIB-mediated C6 oxidation served well for the transformation of commercial L-galactose to the corresponding L-galactosaminuronic acid. Notably, the C6 oxidation of the allyl glycoside was more efficient than that of the selenoglycoside. In addition, a balance between the formation of allyl glycoside and the recovery of selenoglycoside was essential to improve efficiency of the NIS/TfOH-catalyzed allylation. This synthetically useful L-galactosaminuronic acid building block will provide a basis for the syntheses of complex bacterial glycans.
Carbohydrates
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Glycosides
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Oligosaccharides
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Oxidation-Reduction
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Polysaccharides/chemistry*
5.Effects of seedling quality on growth of Rehmannia glutinosa and oligosaccharide content.
Lili ZHOU ; Weizhen YI ; Jianjun QI ; Peng SUN ; Shijie NIU ; Liankui JIAO ; Xian'en LI
China Journal of Chinese Materia Medica 2012;37(14):2037-2040
OBJECTIVETo study the effect of seedling quality on growth, yield and quality of Rehmannia glutinosa at harvest and build a basis for its GAP.
METHODThe seedling quality of R. glutinosa in main producing regions was surveyed to understand the current status of seedling quality. Field experiments with different varieties and seedling quality were conducted to measure dry matter accumulation with different growth of R. glutinosa and oligosaccharide content, and economic yield at harvest.
RESULTThe seedling was randomly selected by farmers in R. glutinosa producing regions. Seedling quality could significantly improve on seedling emergence rate, and promote seedling growth, especially with early stage R. glutinosa, finally increase yield at harvest. At harvest, 63% and 50% of yield with A and B seedling could be improved for variety of 85-5, and 50% and 47% of yield could be increased for variety of Beijing No. 1, compared to the C seeding.
CONCLUSIONIn cultivation, the seedlings with the diameter > 1.5 cm should be transplanted firstly.
Oligosaccharides ; analysis ; Rehmannia ; chemistry ; growth & development ; Seedlings ; chemistry ; physiology
6.Characterization of inulosucrase and the enzymatic synthesis of inulin.
Dawei NI ; Wei XU ; Ziwei CHEN ; Wenli ZHANG ; Wanmeng MU
Chinese Journal of Biotechnology 2021;37(1):266-275
As a type of prebiotics and dietary fiber, inulin performs plenty of significant physiological functions and is applied in food and pharmaceutical fields. Inulosucrase from microorganisms can use sucrose as the substrate to synthesize inulin possessing higher molecular weight than that from plants. In this work, a hypothetical gene coding inulosucrase was selected from the GenBank database. The catalytic domain was remained by N- and C- truncation strategies, constructing the recombinant plasmid. The recombinant plasmid was expressed in E. coli expression system, and after purifying the crude enzyme by Ni²⁺ affinity chromatography, a recombinant enzyme with a molecular weight of approximately 65 kDa was obtained. The optimal pH and temperature of the recombinant enzyme were 5.5 and 45 °C, respectively, when sucrose was used as the sole substrate. The activity of this enzyme was inhibited by various metal ions at different degrees. After purifying the produced polysaccharide, nuclear magnetic resonance analysis was used to determine that the polysaccharide was inulin connected by β-(2,1) linkages. Finally, the conditions for the production of inulin were optimized. The results showed that the inulin production reached the maximum, approximately 287 g/L after 7 h, when sucrose concentration and enzyme dosage were 700 g/L and 4 U/mL, respectively. The conversion rate from sucrose to inulin was approximately 41%.
Escherichia coli/genetics*
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Hexosyltransferases/genetics*
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Inulin
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Oligosaccharides
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Sucrose
7.α-amylase detection methods and applications.
Chinese Journal of Biotechnology 2023;39(3):898-911
α-amylase is an endonucleoside hydrolase that hydrolyzes the α-1, 4-glycosidic bonds inside polysaccharides, such as starch, to generate oligosaccharides, dextrins, maltotriose, maltose and a small amount of glucose. Due to the importance of α-amylase in food industry, human health monitoring and pharmaceuticals, detection of its activity is widely required in the breeding of α-amylase producing strains, in vitro diagnosis, development of diabetes drugs, and the control of food quality. In recent years, many new α-amylase detection methods have been developed with improved speed and sensitivity. This review summarized recent processes in the development and applications of new α-amylase detection methods. The major principle of these detection methods were introduced, and their advantages and disadvantages were compared to facilitate future development and applications of α-amylase detection methods.
Humans
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alpha-Amylases/chemistry*
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Polysaccharides
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Oligosaccharides
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Starch
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Maltose
8.Synthesis of cello-oligosaccharides which promotes the growth of intestinal probiotics by multi-enzyme cascade reaction.
Peng ZHENG ; Lei WANG ; Meirong HU ; Hua WEI ; Yong TAO
Chinese Journal of Biotechnology 2023;39(8):3406-3420
Soluble cello-oligosaccharide with 2-6 oligosaccharide units is a kind of oligosaccharide with various biological functions, which can promote the proliferation of intestinal probiotics such as Bifidobacteria and Lactobacillus paracei. Therefore, it has a regulatory effect on human intestinal microbiota. In this study, a Cc 01 strain was constructed by expressing cellodextrin phosphorylase (CDP) in Escherichia coli. By combining with a previously constructed COS 01 strain, a three-enzyme cascade reaction system based on strains COS 01 and Cc 01 was developed, which can convert glucose and sucrose into cello-oligosaccharide. After optimization, the final titer of soluble cello-oligosaccharides with 2-6 oligosaccharide units reached 97 g/L, with a purity of about 97%. It contained cellobiose (16.8 wt%), cellotriose (49.8 wt%), cellotetrose (16.4 wt%), cellopentaose (11.5 wt%) and cellohexose (5.5 wt%). When using inulin, xylo-oligosaccharide and fructooligosaccharide as the control substrate, the biomass (OD600) of Lactobacillus casei (WSH 004), Lactobacillus paracei (WSH 005) and Lactobacillus acidophilus (WSH 006) on cello-oligosaccharides was about 2 folds higher than that of the control. This study demonstrated the efficient synthesis of cello-oligosaccharides by a three-enzyme cascade reaction and demonstrated that the synthesized cello-oligosaccharides was capable of promoting intestinal microbial proliferation.
Humans
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Oligosaccharides
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Biomass
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Escherichia coli/genetics*
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Gastrointestinal Microbiome
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Glucose
9.Advances in the preparation of alginate oligosaccharides and its biological functions.
Chinese Journal of Biotechnology 2022;38(1):104-118
Alginate is a group of polyuronic saccharides that are widely used in pharmaceutical and food industry due to its unique physicochemical properties and beneficial health effects. However, the low water solubility and high viscosity of alginate hampered its application. Alginate oligosaccharide (AOS) is a decomposition product of alginate and has received increasing attention due to its low molecular weight, high water solubility, safety, and non-toxicity. The wide-ranging biological functions of AOS are closely related to its structural diversity. AOS with distinct structures and biological functions can be obtained by different methods of preparation. This review summarized the biological functions of AOS reported to date, including anti-tumor, immunomodulatory, anti-inflammatory, antioxidant, prebiotic, and anti-diabetes. The preparation of AOS, as well as the relationship between the structure and biological functions of AOS were discussed, with the aim to provide a reference for further development and application of AOS.
Alginates
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Anti-Inflammatory Agents
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Antioxidants
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Molecular Weight
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Oligosaccharides
10.Effect of alginate on early bone healing after implantation of particulate dentin and plaster of paris mixture
Gyung Ahn CHO ; Su Gwan KIM ; Sung Chul LIM ; Sang Gon KIM
Journal of the Korean Association of Maxillofacial Plastic and Reconstructive Surgeons 2005;27(3):218-225
0.05), nor were the differences between the two experimental groups at 4 and 8 weeks. In comparison with the control group, however, both experimental groups showed superior new bone formation. Although the difference between the two experimental groups was not statistically significant, a tendency for superior new bone formation was detected with the combination of particulate dentin, plaster of Paris, and alginate. Furthermore, more fusion of woven bone and bony trabeculae formation occurred in this group, suggesting that oligosaccharides play a role in the formation and maturation of new bone. In conclusion, the treatment of bone defects by grafting with a mixture of particulate dentin and plaster of Paris alone or with alginate significantly improved the formation of new bone, both quantitatively and qualitatively. The oligosaccharide treatment appeared to improve the formation of new bone quantitatively and qualitatively.]]>
Calcium Sulfate
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Dentin
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Free Radicals
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Humans
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Oligosaccharides
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Osteogenesis
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Transplants