Volume 18 Issue 2
Apr.  2022
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YOU Yuxian, XIE Wenyan, BAN Xiaofeng, KONG Haocun, LI Caiming, LI Zhaofeng. Bioinformatics analysis and efficient preparation of β-agarase from marine bacteria[J]. South China Fisheries Science, 2022, 18(2): 1-12. DOI: 10.12131/20210330
Citation: YOU Yuxian, XIE Wenyan, BAN Xiaofeng, KONG Haocun, LI Caiming, LI Zhaofeng. Bioinformatics analysis and efficient preparation of β-agarase from marine bacteria[J]. South China Fisheries Science, 2022, 18(2): 1-12. DOI: 10.12131/20210330
shu

Bioinformatics analysis and efficient preparation of β-agarase from marine bacteria

  • 1.

    School of Food Science and Technology, Jiangnan University, Wuxi 214122, China

  • 2.

    National Engineering Research Center of Cereal Fermentation and Food Biomanufacturing, Wuxi 214122, China

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  • Received Date: November 02, 2021
  • Revised Date: December 26, 2021
  • Accepted Date: January 09, 2022
  • Available Online: January 26, 2022
    Graphical Abstract
    • Abstract
  • In order to expand the high-quality agarase categories, make full use of seaweed resources, and realize the efficient preparation of functional agar oligosaccharides, we derived a β-agarase from the marine bacterium (Agarivorans gilvus WH0801)(β-AGAase) by using genome mining technology, and predicted its physical and chemical properties as well as structural characteristics by bioinformatics analysis. Based on the results, β-AGAase is a non-secreted protein. The β-AGAase was extracellular expressed in Escherichia coli through the introduction of signal peptides by molecular biology methods, and its production efficiency was significantly enhanced through fermentation regulation strategy optimization. The optimal fermentation conditions are as followed: seed culture medium with an inoculation time of 5 h; fermentation starting medium was TB (pH 7.0); carbon source was 6 g·L−1 of fructose; nitrogen source was 30 g·L−1 of yeast extract II. After being cultured at 25 ℃ for 2 h, IPTG was added with a final concentration of 0.025 mmol·L−1 for 48 h induction. Under these conditions, the obtained enzyme activity was 16.72 U·mL−1 and was about 5 times higher than the initial enzyme activity, which proves that β-AGAase is of great potential for industrial application.
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  • [1]
    慕欣. 三株海洋新菌的鉴定及细菌O17~T和TS1~T的琼胶酶研究[D]. 济南: 山东大学, 2017: 1.
    [2]
    王露楠, 杨少玲, 戚勃, 等. 3种改性方法对琼胶理化性质的影响[J]. 南方水产科学, 2021, 17(2): 97-103. doi: 10.12131/20200197
    [3]
    戚勃, 杨贤庆, 李来好, 等. 琼胶寡糖对冻虾仁和罗非鱼片品质的影响[J]. 南方水产科学, 2012, 8(6): 73-79.
    [4]
    XU X Q, SU B M, XIE J S, et al. Preparation of bioactive neoagaroligosaccharides through hydrolysis of Gracilaria lemaneiformi sagar: a comparative study[J]. Food Chem, 2018, 240(1): 330-337.
    [5]
    LEE M H, JANG J H, YOON G Y, et al. Neoagarohexaose-mediated activation of dendritic cells via Toll-like receptor 4 leads to stimulation of natural killer cells and enhancement of antitumor immunity[J]. Bmb Rep, 2017, 50(5): 263-268. doi: 10.5483/BMBRep.2017.50.5.014
    [6]
    WANG W, LIU P, HAO C, et al. Neoagaro-oligosaccharide monomers inhibit inflammation in LPS-stimulated macrophages through suppression of MAPK and NF-κB pathways[J]. Sci Rep, 2017, 7: 44252. doi: 10.1038/srep44252
    [7]
    JOO H S, JE-HYEON L, JOO K E, et al. Anti-obesity and anti-diabetic effect of neoagarooligosaccharides on high-fat diet-induced obesity in mice[J]. Mar Drugs, 2017, 15(4): 90-101. doi: 10.3390/md15040090
    [8]
    HONG S J, LEE J H, KIM E J, et al. In vitro and in vivo investigation for biological activities of neoagarooligosaccharides prepared by hydrolyzing agar with β-agarase[J]. Biotechnol Bioeng, 2017, 22(4): 489-496.
    [9]
    杨绍青, 刘学强, 刘瑜, 等. 酶法制备几种功能性低聚糖的研究进展[J]. 生物产业技术, 2019(4): 10.
    [10]
    ZHANG N, HOU E, SONG J, et al. Neoagarotetraose-modulated gut microbiota and alleviated gut inflammation in antibiotic treatment mice[J]. Food Agr Immunol, 2017, 28(6): 1408-1423. doi: 10.1080/09540105.2017.1346063
    [11]
    YANG M, ZHANG Z L, HE Y, et al. Study on the structure characterization and moisturizing effect of Tremella polysaccharide fermented from GCMCC5.39[J]. Food Sci Hum Well, 2021, 10(4): 471-479. doi: 10.1016/j.fshw.2021.04.009
    [12]
    李静. 琼脂降解菌的筛选及其琼胶酶的研究[D]. 无锡: 江南大学, 2020: 6.
    [13]
    KAZLOWSKI B, PAN C L, KO Y T. Monitoring and preparation of neoagaro- and agaro-oligosaccharide products by high performance anion exchange chromatography systems[J]. Carbohyd Polym, 2015, 122: 351-358. doi: 10.1016/j.carbpol.2014.09.003
    [14]
    ZHANG W, XU J, LIU D, et al. Characterization of an α-agarase from Thalassomonas sp. LD5 and its hydrolysate[J]. Appl Microbiol Biot, 2018, 102(5): 2203-2212. doi: 10.1007/s00253-018-8762-6
    [15]
    林福娣. 新琼寡糖的酶法制备及其生物活性研究[D]. 厦门: 华侨大学, 2020: 2-5.
    [16]
    LI J, XIE M, GAO Y. Identification and biochemical characterization of a novel exo-type β-agarase Aga3463 from an Antarctic Pseudoalteromonas sp. strain[J]. Int J Biol Macromol, 2019, 129: 162-170.
    [17]
    CHEN X, HOU Y, JIN M, et al. Expression and characterization of a novel thermostable and pH-stable β-agarase from deep-sea bacterium Flammeovirga sp. OC4[J]. J Agric Food Chem, 2016, 64(38): 7251-7258. doi: 10.1021/acs.jafc.6b02998
    [18]
    CHEN Y, WU H, WANG G, et al. Inspecting the genome sequence and agarases of Microbulbifer pacificus LD25 from a saltwater hot spring[J]. J Biosci Bioeng, 2019, 127(4): 403-410. doi: 10.1016/j.jbiosc.2018.10.001
    [19]
    CUI X, JIANG Y, CHANG L, et al. Heterologous expression of an agarase gene in Bacillus subtilis, and characterization of the agarase[J]. Int J Biol Macromol, 2018, 120: 657-664. doi: 10.1016/j.ijbiomac.2018.07.118
    [20]
    LI R, YING X, CHEN Z, et al. Expression and characterization of a GH16 family β-agarase derived from the marine bacterium Microbulbifer sp. BN3 and its efficient hydrolysis of agar using raw agar-producing red seaweeds Gracilaria sjoestedtii and Gelidium amansii as substrates[J]. Catalysts, 2020, 10(8): 885.
    [21]
    LEE J, HONG S, LEE C, et al. Production of ethanol from agarose by unified enzymatic saccharification and fermentation in recombinant yeast[J]. J Microbiol Biotech, 2019, 29(4): 625-632. doi: 10.4014/jmb.1902.02012
    [22]
    LI R, CHEN Z, YING X, et al. A novel GH16 beta-agarase isolated from a marine bacterium, Microbulbifer sp. BN3 and its characterization and high-level expression in Pichia pastoris[J]. Int J Biol Macromol, 2018, 119: 1164-1170.
    [23]
    李才明, 黄敏, 石建中, 等. 金属离子协同氨基酸提高重组β-环糊精葡萄糖基转移酶在枯草芽孢杆菌中的表达[J]. 食品与发酵工业, 2016, 42(7): 1-8.
    [24]
    LEE D, PARK G, KIM N, et al. Cloning, expression, and characterization of a glycoside hydrolase family 50 β-agarase from a marine Agarivorans isolate[J]. Biotechnol Lett, 2006, 28(23): 1925-1932. doi: 10.1007/s10529-006-9171-y
    [25]
    DONG J, TAMARU Y, ARAKI T. A unique β-agarase, AgaA, from a marine bacterium, Vibrio sp. strain PO-303[J]. Appl Microbiol Biot, 2007, 74(6): 1248. doi: 10.1007/s00253-006-0781-z
    [26]
    CHEN X, LIN H, JIN M, et al. Characterization of a novel alkaline β-agarase and its hydrolysates of agar[J]. Food Chem, 2019, 295(15): 311-319.
    [27]
    班宵逢. 静电相互作用对淀粉分支酶热稳定性影响的研究[D]. 无锡: 江南大学, 2017: 50.
    [28]
    杨平, 张边江, 王立科, 等. 豆科家族中的木糖异构酶基因分析[J]. 食品与发酵工业, 2020, 46(19): 23-27.
    [29]
    杨艳北, 许晶, 沈城辉, 等. N-酰基高丝氨酸内酯酶的生物信息学分析[J]. 甘肃农业科技, 2021, 52(2): 31-37.
    [30]
    李兆丰. 软化类芽孢杆菌α-环糊精葡萄糖基转移酶在大肠杆菌中的表达及其产物特异性分析[D]. 无锡: 江南大学, 2009: 22-25.
    [31]
    LIU J, LIU Z, JIANG C, et al. Biochemical characterization and substrate degradation mode of a novel α-agarase from Catenovulum agarivorans[J]. J Agric Food Chem, 2019, 67(37): 10373-10379. doi: 10.1021/acs.jafc.9b03073
    [32]
    DONG Q, RUAN L W, SHI H. A β-agarase with high pH stability from Flammeovirga sp. SJP92[J]. Carbohydr Res, 2016, 432(2): 1-8.
    [33]
    文霞, 周少璐, 杨秀茳, 等. 海洋微生物多糖降解酶的研究进展[J]. 生物技术通报, 2016, 32(11): 38-46.
    [34]
    王晨, 赵雨佳, 李春, 等. 动态转录调控微生物代谢途径研究进展[J]. 化工进展, 2019, 38(9): 4238-4246.
    [35]
    冉凡娇, 孙谧, 包静, 等. Bohai sea-9145重组脂肪酶基因工程菌的发酵表达条件优化[J]. 食品工业科技, 2016, 37(16): 178-183.
    [36]
    韩来闯. 生物元件的挖掘、改造及在基因表达调控系统中的应用[D]. 无锡: 江南大学, 2020: 6.
    [37]
    于平, 陈凯飞, 朱祺, 等. 重组大肠杆菌生物合成γ-氨基丁酸的发酵条件优化[J]. 中国食品学报, 2018, 18(6): 112-120.
    [38]
    ZHU Y, ZHAO R, XIAO A, et al. Characterization of an alkaline β-agarase from Stenotrophomonas sp. NTa and the enzymatic hydrolysates[J]. Int J Biol Macromol, 2016, 86: 525-534. doi: 10.1016/j.ijbiomac.2016.01.106
    [39]
    RAMOSA K R M, VALDEHUESAA K N G, NISOLA G M, et al. Identification and characterization of a thermostable endolytic β-agarase Aga2 from a newly isolated marine agarolytic bacteria Cellulophaga omnivescoria W5C[J]. New Biotechnol, 2018, 40: 261-267. doi: 10.1016/j.nbt.2017.09.006
    [40]
    AN K, SHI X, CUI F. Characterization and overexpression of a glycosyl hydrolase family 16 beta-agarase YM01-1 from marine bacterium Catenovulum agarivorans YM01T[J]. Protein Expres Purif, 2018, 143: 1-8. doi: 10.1016/j.pep.2017.10.002
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