glnA基因敲除对土壤杆菌产可得然胶的影响-食品安全质量检测学报-CSCIED科技核心评价数据库-手机版

glnA基因敲除对土壤杆菌产可得然胶的影响

Effects of glnA gene knockout on the production of available curdlan by Agrobacterium

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DOI	10.19812/j.cnki.jfsq11-5956/ts.20250120006
刊名	食品安全质量检测学报 Journal of Food Safety and Quality
年，卷(期)	2025, 16(9)
作者	姚博伟, 武世强, 高红亮, 杨雪霞,
作者单位	宁夏计量质量检验检测研究院; 东华大学生物与医学工程学院; 华东师范大学生命科学学院
摘要	目的探究glnA基因对土壤杆菌产可得然胶的影响。方法构建glnA基因敲除株ΔglnA菌株, 分析ΔglnA菌株生长代谢情况和产生的可得然胶在产量、凝胶性质和红外结构的变化, 并通过实时荧光定量聚合酶链式反应的方法探究可得然胶合成相关基因的表达量。结果 ΔglnA菌株在发酵过程中氨基氮消耗情况与CGMCC11546一致, 在蔗糖消耗方面, ΔglnA菌株与CGMCC11546在12 h之前无明显差别, 但在15 h之后出现明显差异, 蔗糖消耗比CGMCC11546明显降低。ΔglnA菌株可得然胶产量约4 g/L, 相对于CGMCC11546降低了60%。结论 glnA基因不影响菌体生长和可得然胶结构, 但可以使可得然胶的合成减少。本研究通过对glnA基因进行敲除, 研究其对可得然胶合成的影响, 为后续提高可得然胶产量提供一些分子生物学基础。
Abstract	Objective To explore the effects of glnA gene on the production of curdlan by Agrobacterium. Methods A glnA gene knockout strain ΔglnA was constructed in this paper. The growth and metabolism of ΔglnA strain and the changes in yield, gel properties and infrared structure of the obtained gum were analyzed, and the expression of genes related to the synthesis of curdlan was explored by quantitative real-time polymerase chain reaction. Results The amino nitrogen consumption of the ΔglnA strain during fermentation was consistent with that of CGMCC11546. In terms of sucrose consumption, there was no significant difference before 12 h, but there was a significant difference between the ΔglnA strain and CGMCC11546 after 15 h, and sucrose consumption was significantly reduced compared to CGMCC11546. The ΔglnA strain could produce approximately 4 g/L of natural gum, which was 60% lower than CGMCC11546. Conclusion The glnA gene does not affect bacterial growth or the structure of collagen, but it can reduce the synthesis of collagen. This study provides insights into the role of glnA in curdlan production and lays a molecular biological foundation for further efforts to enhance curdlan yield.
关键词	可得然胶;土壤杆菌;基因敲除
KeyWord	curdlan; Agrobacterium; gene knockout; glnA gene
基金项目
页码	33-39

参考文献
相关文献

[1]HARADA T, FUJIMORI K, HIROSE S, et al. Growth and β-glucan 10C3K production by a mutant of Alcaligenes faecalis var. myxogenes in defined medium
[J]. Microbiogy and Fermentation Industry, 1966, 30(8): 764–769.
[2]PHILLIPS KR, PIK J, LAWFORD HG, et al. Production of curdlan-type polysaccharide by Alcaligenes faecalis in batch and continuous culture
[J]. Canadian Journal of Microbiology, 1983, 29(10): 1331–1338.
[3]NAKANISHI I, KIMURA K, SUZUKI T, et al. Demonstration of curdlan-type polysaccharide and some other P-1,3-glucan in microorganisms with aniline blue
[J]. The Journal of General and Applied Microbiology, 2006, 22(1): 1–11.
[4]杨镕, 臧一宇, 吴鹏, 等. 多糖类食品胶体的功能特性及其在食品加工中的应用研究进展
[J]. 食品科学, 2024, 45(5): 283–292. YANG R, ZANG YY, WU P, et al. Research progress on functional properties of polysaccharide-based food colloids and their application in food processing
[J]. Food Science, 2024, 45(5): 283–292.
[5]CHEN Y, BERA H, SI L, et al. Tailor-made curdlan based nanofibrous dressings enable diabetic wound healing
[J]. Carbohydrate Polymers, 2025, 348(PB): 122876.
[6]MOUSA I, ALSHARARI MA, ALALAWY IA, et al. Harnessing the potential of sargassum latifolium for sustainable synthesis of Ag/Bi2O3-curdlan nanocomposites: Phytochemical characterization and biological applications
[J]. Journal of Inorganic and Organometallic Polymers and Materials, 2025. https://doi.org/10.1007/s10904-024- 03566-7
[7]ZHUANG YD, FAN CQ, ZHENG YY, et al. Study on the interaction properties between curdlan polysaccharide and water molecules in coal processing sludge by molecular dynamics simulation
[J]. Journal of Nanoparticle Research, 2024, 26(7): 163.
[8]HONGLIANG G, LI Y, JIANGTAO T, et al. Characterization and rheological properties analysis of the succinoglycan produced by high-yield mutant of Rhizobium radiobacter ATCC 19358
[J]. International Journal of Biological Macromolecules, 2021, 166: 61–70.
[9]MENG Y, GANG F, YUMEI S, et al. Biosynthesis and applications of curdlan
[J]. Carbohydrate Polymers, 2021, 273: 118597.
[10]TAKASHI K, NOBUYUKI S, KOJI N. Heat-melt of β-1,3-D-glucan gel
[J]. Agricultural & Biological Chemistry, 1977, 41(7): 1315–1316.
[11]ZHANG H, NISHINARI K, WILLIAMS MAK, et al. A molecular description of the gelation mechanism of curdlan
[J]. International Journal of Biological Macromolecules, 2002, 30(1): 7–16.
[12]CHO SS. Handbook of dietary fiber
[M]. Boca Raton: CRC Press, 2001.
[13]VERMA DK, NIAMAH AK, PATEL AR, et al. Chemistry and microbial sources of curdlan with potential application and safety regulations as prebiotic in food and health
[J]. Food Research International, 2020, 133(prepublish): 109136.
[14]曹云刚, 胡春杰, 李远征, 等. 可得然胶功能特性及在肉及肉制品中的应用进展
[J]. 精细化工, 2024, 41(3): 491–498, 587. CAO YG, HU CJ, LI YZ, et al. Functional properties of curdlan and its application progress in meat and meat products
[J]. Fine Chemicals, 2024, 41(3): 491–498, 587.
[15]HOU L, ZHAO B, FU S, et al. Effects of curdlan on the quality of frozen steamed bread: Quality changes, water state and starch crystallinity
[J]. International Journal of Food Science & Technology, 2024, 59(9): 6611–6618.
[16]VASIEE A, JAMAB SM, FALAH F, et al. Microbial production of curdlan in sugar beet molasses medium: Effects on physicochemical attributes of reduced-fat frankfurter sausages
[J]. LWT-Food Science and Technology, 2025, 216: 117310.
[17]BIELECKA MM, FLORCZUK A, ALJEWICZ M. An evaluation of the impact of curdlan and buttermilk addition on the functional properties and sensory quality of processed cheese analogs
[J]. Molecules, 2024, 30(1): 66.
[18]LU M, ZHANG C, MA L, et al. Cryoprotective effect of magnetic field-assisted freezing in combination with curdlan on myofibrillar protein of Litopenaeus vannamei: Oxidative aggregation, protein structure and thermal stability
[J]. Food Hydrocolloids, 2025, 160(P1): 110730.
[19]SUN X, LIU Y, LI X, et al. Curdlan inclusion modifies the rheological properties and the helix-coil transition behavior of gelatin and increases the flexibility of gelatin films
[J]. Food Chemistry, 2025, 469: 142567.
[20]CHEN K, JIANG J, TIAN Y, et al. Improved konjac glucomannan/curdlan-based emulsion coating by mung bean protein addition for cherry tomato preservation
[J]. International Journal of Biological Macromolecules, 2024, 291: 139080.
[21]MOON CJ, LEE JH. Use of curdlan and activated carbon composed adsorbents for heavy metal removal
[J]. Process Biochemistry, 2005, 40(3/4): 1279–1283.
[22]张静. 土壤杆菌合成可得然胶相关基因的筛选及其分子机制研究
[D]. 上海: 华东师范大学, 2019. ZHANG J. The research of screening and molecular mechanism of genes involved in curdlan synthesis in Agrobacterium sp.
[D]. Shanghai: East China Normal University, 2019.
[23]LEE SY. Bacterial polyhydroxyalkanoates
[J]. Biotechnology and Bioengineering, 1996, 49(1): 1–14.
[24]WOJCIECH S, EWELINA J, DUSAN M, et al. Physicochemical studies of novel sugar fatty acid esters based on (R)-3-hydroxylated acids derived from bacterial polyhydroxyalkanoates and their potential environmental impact
[J]. Frontiers in Bioengineering and Biotechnology, 2023, 11: 1112053.
[25]CHARLES TC, CAI GQ, ANEJA P. Megaplasmid and chromosomal loci for the PHB degradation pathway in Rhizobium (Sinorhizobium) meliloti
[J]. Genetics, 1997, 146(4): 1211–1220.
[26]武世强, 蒋德明, 张静, 等. phbC基因敲除对土壤杆菌产可得然胶的影响
[J]. 工业微生物, 2019, 49(4): 21–26. WU SQ, JIANG DM, ZHANG J, et al. The effect of phbC gene knockout on the production of available curdlan by Agrobacterium
[J]. Industrial Microbiology, 2019, 49(4): 21–26.
[27]ZHANG HT, ZHAN XB, ZHENG ZY, et al. Sequence and transcriptional analysis of the genes responsible for curdlan biosynthesis in Agrobacterium sp. ATCC 31749 under simulated dissolved oxygen gradients conditions
[J]. Applied Microbiology and Biotechnology, 2011, 91(1): 163–175.
[28]STASINOPOULOS SJ, FISHER PR, STONE BA, et al. Detection of two loci involved in (1→3)-β-glucan (curdlan) biosynthesis by Agrobacterium sp. ATCC31749, and comparative sequence analysis of the putative curdlan synthase gene
[J]. Glycobiology, 1999(1): 31–41.
[29]TARA K, CHANDANA VE, BRUCE ASA, et al. Topological characterization of an inner membrane (1→3)-β-D-glucan (curdlan) synthase from Agrobacterium sp. strain ATCC 31749
[J]. Glycobiology, 2003, 13(10): 693–-706.
[30]YU X, ZHANG C, YANG L, et al. CrdR function in a curdlan-producing Agrobacterium sp. ATCC31749 strain
[J]. BMC Microbiology, 2015, 15(1): 25.
[31]MCINTOSH M, STONE BA, STANISICH VA. Curdlan and other bacterial (1→3)-β-d-glucans
[J]. Applied Microbiology and Biotechnology, 2005, 68(2): 163–173.
[32]WU JR, YU LJ, ZHAN XB, et al. NtrC-dependent regulatory network for curdlan biosynthesis in response to nitrogen limitation in Agrobacterium sp. ATCC 31749
[J]. Process Biochemistry, 2012, 47(11): 1552–1558.
[33]YU L, WU J, LIU J, et al. Enhanced curdlan production in Agrobacterium sp. ATCC 31749 by addition of low-polyphosphates
[J]. Biotechnology & Bioprocess Engineering, 2011, 16(1): 34–41.
[34]YU LJ, WU JR, ZHENG ZZ, et al. Changes in gene transcription and protein expression involved in the response of Agrobacterium sp. ATCC 31749 to nitrogen availability during curdlan production
[J]. Applied Biochemistry & Microbiology, 2011, 47(5): 487–493.
[35]张靖宇. 农杆菌可得然胶合成相关双组分调节系统基因的筛选与机制探究
[D] 上海: 华东师范大学, 2023. ZHANG JY. Screening and regulation mechanism of twocomponent systems of Agrobacterium related to curdlan biosynthesis
[D]. Shanghai: East China Normal University, 2023.
[36]于婷婷, 韩鸿宇, 张永刚, 等. 高凝胶强度可得然胶生产菌的构建筛选与发酵工艺优化研究
[J]. 食品与发酵工业, 2024, 50(18): 82–90. YU TT, HAN HY, ZHANG YG, et al. Construction and screening of high gel strength curdlan production bacteria and optimization of fermentation medium
[J]. Food and Fermentation Industries, 2024, 50(18): 82–90.
[37]YU T, WANG Y, WANG W, et al. Optimizing curdlan synthesis: Engineering Agrobacterium tumefaciens ATCC31749 for enhanced production using dextrin as a carbon source
[J]. Fermentation, 2024, 10(5): 240.
[38]刘晓霞. 土壤杆菌产可得然胶发酵工艺优化及其性质的研究
[D]. 上海: 华东师范大学, 2017. LIU XX. The research of fermentation process optimization and properties of curdlan by Agrobacterium sp.
[D]. Shanghai: East China Normal University, 2017.
[39]ZHANG W, GAO HL, HUANG YM, et al. Glutamine synthetase gene glnA plays a vital role in curdlan biosynthesis of Agrobacterium sp. CGMCC 11546
[J]. International Journal of Biological Macromolecules, 2020, 165(PA): 222–230.
[40]俞珊, 段孟霞, 童彩玲, 等. 可得然胶功能性质及其在食品中的应用研究进展
[J]. 食品科学, 2022, 43(19): 277–284. YU S, DUAN MX, TONG CL, et al. Progress in research on functional properties of curdlan and its application in food
[J]. Food Science, 2022, 43(19): 277–284.
[41]AMBROOK J, DAVID RUSSELL. 分子克隆实验指南
[M]. 北京: 科学出版社, 2002. SAMBROOK J, DAVID R. Molecular cloning: A laboratory manual
[M]. Beijing: Science Publishing Company, 2002.

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姚博伟*, 武世强, 高红亮, 杨雪霞*. glnA基因敲除对土壤杆菌产可得然胶的影响 [J]. 食品安全质量检测学报. 2025; 16; (9). 33 - 39.

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