纳豆芽胞杆菌固定化载体改性及吸附研究

陈龙军; 林程; 林陈强; 张慧; 贾宪波; 方宇; 陈济琛

doi:10.19303/j.issn.1008-0384.2021.07.015

纳豆芽胞杆菌固定化载体改性及吸附研究

doi: 10.19303/j.issn.1008-0384.2021.07.015

陈龙军^1,,
林程²,
林陈强¹,
张慧¹,
贾宪波¹,
方宇¹,
陈济琛^1, ,

1.
福建省农业科学院土壤肥料研究所，福建　福州　350003
2.
福州市农业农村局，福建　福州　350003

基金项目: 福建省农业科学院探索创新项目（AA2018-13）；福建省科技计划区域发展项目（2019Y3005）；福建省科技计划公益类专项（2018R1022-7）、（2019R1025-4）；国家重点研发计划项目（2018YFD0200706）

详细信息

作者简介:
陈龙军（1983−），男，硕士，助理研究员，主要从事微生物菌剂、酶工程及发酵工程研究（E-mail：monkeyirene@163.com）

通讯作者:
陈济琛（1964−），男，研究员，主要从事微生物菌剂研究（E-mail：chenjichen2001@163.com）

中图分类号: Q 939.96
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出版历程
- 收稿日期: 2020-07-22
- 修回日期: 2021-07-02
- 网络出版日期: 2021-07-13
- 刊出日期: 2021-07-28

Functional Improvements on Immobilization Carrier of Bacillus Culture for Natto Fermentation

1.
Soil and Fertilizer Institute, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian　350003, China
2.
Fuzhou Agricultural and Rural Bureau, Fuzhou, Fujian　350003, China

摘要

摘要: 目的以麦饭石为载体，制备高效的纳豆芽胞杆菌固体菌剂，为纳豆芽胞杆菌的规模化应用奠定基础。方法运用酸、碱、盐等方法探索麦饭石进行改性条件，扩大麦饭石孔隙度及比表面积，提升其吸附能力；并通过添加保护剂（葡萄糖、糊精、淀粉）增加保藏期固体菌剂的有效活菌数残留率，为长期保藏固体粉剂提供技术支持。结果（1）确定了纳豆芽胞杆菌最适培养温度为40 ℃，最适培养pH为7.5。（2）经氯化钠、盐酸、硫酸和氢氧化钠溶液改性后的麦饭石吸附最大有效活菌数分别为13.17、12.6、12.7和11.9（×10⁸ CFU·g⁻¹）。其中0.5 mol·L⁻¹氯化钠溶液改性效果最好，吸附有效活菌数达到13.17（×10⁸ CFU·g⁻¹）。（3）对照组、添加保护剂组（2%糊精、2%葡萄糖、2%淀粉）保藏至6个月时，有效活菌数分别为2.91、4.1、5.42和4.63（×10⁸ CFU·g⁻¹）。活菌数残留率为初始活菌数的22%、31%、41%和35%，其中2%葡萄糖保护剂比对照组有效活菌数残留率提高了1.86倍。结论改性麦饭石能极大提升麦饭石的吸附能力，同时保护剂的添加有利于麦饭石固体芽胞杆菌粉剂的长期保藏，本研究为麦饭石在微生物菌剂制备工艺的应用提供了一定的技术支持。
- 纳豆芽胞杆 /
- 麦饭石 /
- 改性 /
- 保护剂 /
- 吸附
Abstract: Objective The physical properties of maifanshi, a volcanic rock, were modified to improve its porosity and adsorption to be used as an immobilization carrier for natto fermentation bacteria that was further added with protective agents to extend the shelf life of viable Bacillus natto. Method Maifanshi was treated by acid, alkali, or salt to modify the porosity and surface adsorption property as a microbial carrier. Various protective agents, such as glucose, dextrin, and/or starch were incorporated to help prolong the viability of the imbedded B. natto powder. Result (1) The optimum conditions for B. natto culture included 40 ℃ and pH 7.5. (2) The maximum viable bacteria count was the highest at 13.17×10⁸ CFU·g⁻¹ on the maifanshi carriers modified by 0.5 mol·L⁻¹sodium chloride solution. It was followed by a count of 12.7 ×10⁸ CFU·g⁻¹ on maifanshi treated by sulfuric acid, 12.6×10⁸ CFU·g⁻¹ on maifanshi treated by hydrochloric acid, and 11.9×10⁸ CFU·g⁻¹ on maifanshi treated by sodium hydroxide. (3) The viable B. natto counts after 6 months in storage on control was 2.91×10⁸ CFU·g⁻¹ as compared to those immobilized on the carriers with addition of 2% dextrin, 2% glucose, or 2% starch being 4.1×10⁸ CFU·g⁻¹, 5.42×10⁸ CFU·g⁻¹, and 4.63×10⁸ CFU·g⁻¹, respectively. The retention of viable B. natto on the NaCl-treated maifanshi with the addition of dextrin was 31%, that of glucose 41%, that of starch 35%, and that of control 22%. Conclusion As a bacteria carrier, maifanshi could be significantly further improved on the adsorption capacity by modification such as NaCl treatment. With an addition of 2% glucose on the immobilized system, the retained viable B. natto count could rise 1.86 times over control in 6 months of storage. It appeared that this study had paved the way of developing a practical application of maifanshi for natto fermentation industry.
- Bacillus natto /
- maifanshi /
- modification /
- protectant /
- adsorption

HTML全文

图 1 不同培养温度对纳豆芽胞杆菌有效活菌数的影响

Figure 1. Effect of culture temperature on viable B. natto count

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图 2 不同培养pH对纳豆芽胞杆菌有效活菌数的影响

Figure 2. Effect of pH on viable B. natto count

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图 3 不同改性剂及浓度对纳豆芽胞杆菌吸附效果的影响

Figure 3. Adsorption of maifanshi affected by type and concentration of modifiers

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图 4 吸附纳豆芽胞杆菌在扫描电镜下观察图

注：A为改性前；B为改性后；C为改性吸附后。

Figure 4. Scanning electron microscope images of B. natto on maifanshi

Note: A: unmodified; B: modified; C: adsorption after modification.

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图 5 不同保护剂对纳豆芽胞杆菌有效活菌数的影响

Figure 5. Effects of protectants on viable B. natto count

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表 1 麦饭石改性前后吸附效果比较

Table 1. Maifanshi adsorption property after modification treatments

项目 Item	天然麦饭石 Natural Maifan stone	改性麦饭石 Modified Maifan stone
改性条件 Modification conditions	未改性 Unmodified	0.5 mol·L⁻¹ NaCl溶液 0.5 mol·L⁻¹ NaCl solution
吸附有效活菌数 Amount of bacteria absorbed/ （×10⁸ CFU·g⁻¹）	11.35±0.35	13.17±0.44
初始纳豆芽胞杆菌 Bacillus natrius/ （×10⁸ CFU·mL⁻¹）	13.68	13.68
吸附率 Adsorption rate/%	83	96.3
注：数据为平均值 ± 标准差。 Note: Data are mean±standard deviation.

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参考文献(17)

[1]	林先贵, 胡君利. 土壤微生物多样性的科学内涵及其生态服务功能 [J]. 土壤学报, 2008, 45(5):892−900. doi: 10.3321/j.issn:0564-3929.2008.05.016 LIN X G, HU J L. Scientific connotation and ecological service function of soil microbial diversity [J]. Acta Pedologica Sinica, 2008, 45(5): 892−900.(in Chinese) doi: 10.3321/j.issn:0564-3929.2008.05.016
[2]	许杰龙, 张国霞, 许玫英, 等. 餐厨废弃物资源化利用的微生物技术研究进展 [J]. 微生物学通报, 2011, 38(6):928−933. XU J L, ZHANG G X, XU M Y, et al. Research progress on reuse of food waste with microbial technology [J]. Microbiology China, 2011, 38(6): 928−933.(in Chinese)
[3]	THATOI H, BEHERA B C, MISHRA R R, et al. Biodiversity and biotechnological potential of microorganisms from mangrove ecosystems: A review [J]. Annals of Microbiology, 2013, 63(1): 1−19. doi: 10.1007/s13213-012-0442-7
[4]	朱永光, 杨柳, 张火云, 等. 微生物菌剂的研究与开发现状 [J]. 四川环境, 2004, 23(3):5−8, 12. doi: 10.3969/j.issn.1001-3644.2004.03.002 ZHU Y G, YANG L, ZHANG H Y, et al. Progress of research and development of microorganisms agents for bioaugmentation [J]. Sichuan Environment, 2004, 23(3): 5−8, 12.(in Chinese) doi: 10.3969/j.issn.1001-3644.2004.03.002
[5]	李静, 谭月臣, 洪剑明. 固定化微生物技术处理废水 [J]. 安徽农业科学, 2010, 38(25):13981−13983. doi: 10.3969/j.issn.0517-6611.2010.25.161 LI J, TAN Y C, HONG J M. Use of microorganism immobilization technology in wastewater treatment [J]. Journal of Anhui Agricultural Sciences, 2010, 38(25): 13981−13983.(in Chinese) doi: 10.3969/j.issn.0517-6611.2010.25.161
[6]	张陇利, 刘青, 徐智, 等. 复合微生物菌剂对污泥堆肥的作用效果研究 [J]. 环境工程学报, 2008, 2(2):266−269. ZHANG L L, LIU Q, XU Z, et al. Research on effect of compound microbial inoculant on sewage sludge composting [J]. Chinese Journal of Environmental Engineering, 2008, 2(2): 266−269.(in Chinese)
[7]	勾长龙, 高云航, 刘淑霞, 等. 微生物菌剂对堆肥发酵影响的研究进展 [J]. 湖北农业科学, 2013, 52(6):1244−1247. doi: 10.3969/j.issn.0439-8114.2013.06.002 GOU C L, GAO Y H, LIU S X, et al. Investigation and application progress of inoculating microbes on aerobic composting of manure [J]. Hubei Agricultural Sciences, 2013, 52(6): 1244−1247.(in Chinese) doi: 10.3969/j.issn.0439-8114.2013.06.002
[8]	官赟赟, 顾锡慧, 雷太平, 等. 炼油废水COD降解菌固态菌剂的制备及性能考察 [J]. 工业水处理, 2016, 36(2):75−78. doi: 10.11894/1005-829x.2016.36(2).075 GUAN Y Y, GU X H, LEI T P, et al. Study on the preparation and performances of solid bacteria agent of COD degrading bacteria of refinery wastewater [J]. Industrial Water Treatment, 2016, 36(2): 75−78.(in Chinese) doi: 10.11894/1005-829x.2016.36(2).075
[9]	王微, 李圆萍, 谢惠玲, 等. 铜绿假单胞菌菌剂载体的筛选 [J]. 福建农林大学学报(自然科学版), 2016, 45(3):325−330. WANG W, LI Y P, XIE H L, et al. Screening of carrier for Pseudomonas aeruginosa bacterial agent [J]. Journal of Fujian Agriculture and Forestry University (Natural Science Edition), 2016, 45(3): 325−330.(in Chinese)
[10]	赵旭, 王改兰, 陈春玉, 等. 谷糠和堆肥作为微生物菌剂载体的研究 [J]. 安徽农业科学, 2015, 43(14):179−180, 223. doi: 10.3969/j.issn.0517-6611.2015.14.069 ZHAO X, WANG G L, CHEN C Y, et al. Study of bran and compost as a carrier for microbial inoculants [J]. Journal of Anhui Agricultural Sciences, 2015, 43(14): 179−180, 223.(in Chinese) doi: 10.3969/j.issn.0517-6611.2015.14.069
[11]	高书锋, 刘惠知, 胡新旭, 等. 畜禽用凝结芽孢杆菌菌剂制备工艺的初步研究 [J]. 饲料工业, 2012, 33(13):37−40. doi: 10.3969/j.issn.1001-991X.2012.13.009 GAO S F, LIU H Z, HU X X, et al. Preliminary researches on preparation techology of Bacillus coagulan strain applied to animal [J]. Feed Industry, 2012, 33(13): 37−40.(in Chinese) doi: 10.3969/j.issn.1001-991X.2012.13.009
[12]	李娟, 张盼月, 高英, 等. 麦饭石的理化性能及其在水质优化中的应用 [J]. 环境科学与技术, 2008, 31(10):63−66, 75. doi: 10.3969/j.issn.1003-6504.2008.10.016 LI J, ZHANG P Y, GAO Y, et al. Overview of Maifanshi: its physi-chemical properties and nutritious function in drinking water [J]. Environmental Science & Technology, 2008, 31(10): 63−66, 75.(in Chinese) doi: 10.3969/j.issn.1003-6504.2008.10.016
[13]	陈丽红, 张翔凌, 何春艳, 等. Zn-LDHs覆膜改性麦饭石对Cd(Ⅱ)吸附性能及其作用机理研究 [J]. 环境科学学报, 2019, 39(12):4004−4014. CHEN L H, ZHANG X L, HE C Y, et al. Adsorption behavior and mechanisms for Cd(Ⅱ) by modified maifanite coated with Zn-LDHs [J]. Acta Scientiae Circumstantiae, 2019, 39(12): 4004−4014.(in Chinese)
[14]	陈琳荔, 邹华. 改性麦饭石对水中氮磷的去除 [J]. 食品与生物技术学报, 2015, 34(3):283−290. CHEN L L, ZOU H. Experimental study on removal of ammonium and phosphate in simulation river water by modified Maifan tone [J]. Journal of Food Science and Biotechnology, 2015, 34(3): 283−290.(in Chinese)
[15]	张琪. 枯草芽孢杆菌固定化载体的改性条件及处理模拟污水[D]. 哈尔滨: 哈尔滨理工大学, 2014. ZHANG Q. The modified conditions of Bacillus subtilis immobilized carrier and the treatment of simulated sewage[D]. Harbin: Harbin University of Science and Technology, 2014. (in Chinese).
[16]	陈龙军, 林陈强, 张慧, 等. 纳豆芽胞杆菌Bacillus natto菌剂载体的筛选 [J]. 福建农业学报, 2018, 33(10):1104−1107. CHEN L J, LIN C Q, ZHANG H, et al. Selection of carrier for dried Bacillus natto [J]. Fujian Journal of Agricultural Sciences, 2018, 33(10): 1104−1107.(in Chinese)
[17]	常金梅, 蔡芷荷, 吴清平, 等. 菌种冷冻干燥保藏的影响因素 [J]. 微生物学通报, 2008, 35(6):959−962. doi: 10.3969/j.issn.0253-2654.2008.06.024 CHANG J M, CAI Z H, WU Q P, et al. Influencing factors to freeze-drying preservation of culture [J]. Microbiology, 2008, 35(6): 959−962.(in Chinese) doi: 10.3969/j.issn.0253-2654.2008.06.024