烷基季铵化壳聚糖的制备及其抗生物被膜的活性

doi:10.16085/j.issn.1000-6613.2022-1610

化工进展 ›› 2023, Vol. 42 ›› Issue (4): 1995-2002.DOI: 10.16085/j.issn.1000-6613.2022-1610

烷基季铵化壳聚糖的制备及其抗生物被膜的活性

王林¹(), 辛梅华¹(), 李明春¹(), 张涛¹, 毛扬帆²

^1.华侨大学材料科学与工程学院，环境友好功能材料教育部工程中心，福建厦门 361021
^2.华侨大学分析测试中心，福建厦门 361021

收稿日期:2022-08-31 修回日期:2022-10-20 出版日期:2023-04-25 发布日期:2023-05-08
通讯作者: 辛梅华,李明春
作者简介:王林（1994—），男，博士研究生，研究方向为功能高分子材料。E-mail: 1107126210@qq.com。
基金资助:
福建省科技计划(2019H6014)

Preparation of alkyl quaternized chitosan and its anti-biofilm activity

WANG Lin¹(), XIN Meihua¹(), LI Mingchun¹(), ZHANG Tao¹, MAO Yangfan²

^1.Engineering Research Center of Environment-Friendly Functional Materials, Ministry of Education, College of Material Science and Engineering, Huaqiao University, Xiamen 361021, Fujian, China
^2.The Instrumental Analysis Center, Huaqiao University, Xiamen 361021, Fujian, China

Received:2022-08-31 Revised:2022-10-20 Online:2023-04-25 Published:2023-05-08
Contact: XIN Meihua, LI Mingchun

摘要/Abstract

摘要：

通过两步反应制备了4种不同烷基链长的壳聚糖季铵盐，包括N,N,N-三甲基苄基壳聚糖季铵盐（TMBC）、N,N-二甲基-N-丁基苄基壳聚糖季铵盐（DMBBC）、N,N-二甲基-N-辛基壳聚糖季铵盐（DMOBC）和N,N-二甲基-N-十二烷基苄基壳聚糖季铵盐（DMDBC）。采用¹H NMR和TG对产物进行表征，并对其体外抗菌活性和抗细菌生物被膜活性进行测试。结果表明，烷基链越长，壳聚糖季铵盐的抗菌活性越强，DMDBC对E. coli的MIC和MBC分别为0.156mg/mL和0.625mg/mL，对S. aureus的MIC和MBC分别为0.039mg/mL和0.156mg/mL。浓度为0.5mg/mL时对E. coli和S. aureus的抑菌率均达到100%。在1/2MIC浓度时对E. coli和S. aureus的BF形成抑制率分别为76.43%和76.96%，浓度2.5mg/mL时对E. coli和S. aureus的成熟生物被膜清除率分别为67.60%和92.93%。

关键词: 壳聚糖季铵盐, 生物被膜, 抗菌活性

Abstract:

Four chitosan quaternary ammonium salts with different alkyl chain lengths were successfully synthesized, including N,N,N-trimethylbenzyl chitosan chloride (TMBC), N,N-dimethyl-N-butyl benzyl chitosan chloride (DMBBC), N,N-dimethyl-N-octyl benzyl chitosan chloride (DMOBC) and N,N-dimethyl-N-dodecyl benzyl chitosan chloride (DMDBC), and characterized by nuclear magnetic resonance (¹H NMR) spectra and thermogravimetric analysis (TGA). Their antibacterial activities against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) were investigated in vitro using the antibacterial assay, minimal inhibitory concentration and minimum bactericidal concentration assay, while antibiofilm activity was quantified by crystal violet staining assay. The antibacterial activity revealed that the alkyl chain length could affect the antibacterial effect of chitosan quaternary ammonium salt. Especially, DMDBC exhibited the best antibacterial property. The MIC and MBC of DMDBC for E. coli were 0.156mg/mL and 0.625mg/mL, respectively, and for S. aureus were 0.039mg/mL and 0.156mg/mL, respectively. At 0.5mg/mL, the antibacterial rate of DMDBC against E. coli and S. aureus could reach 100%. At 1/2 MIC, the inhibition rates of BF formation of E. coli and S. aureus by DMDBC were 76.43% and 76.96%, respectively, and at 2.5mg/mL, the mature biofilm clearance rates of E. coli and S. aureus were 67.60% and 92.93%, respectively.

Key words: chitosan quaternary ammonium salt, biofilm, antibacterial

中图分类号:

O636.1

王林, 辛梅华, 李明春, 张涛, 毛扬帆. 烷基季铵化壳聚糖的制备及其抗生物被膜的活性[J]. 化工进展, 2023, 42(4): 1995-2002.

WANG Lin, XIN Meihua, LI Mingchun, ZHANG Tao, MAO Yangfan. Preparation of alkyl quaternized chitosan and its anti-biofilm activity[J]. Chemical Industry and Engineering Progress, 2023, 42(4): 1995-2002.

图/表 11

参考文献 17

1	IRIBARNEGARAY Victoria, NAVARRO Nicolas, ROBINO Luciana, et al. Magnesium-doped zinc oxide nanoparticles alter biofilm formation of Proteus mirabilis[J]. Nanomedicine, 2019, 14(12): 1551-1564.
2	QUAN Kecheng, ZHANG Zexin, CHEN Hong, et al. Artificial channels in an infectious biofilm created by magnetic nanoparticles enhanced bacterial killing by antibiotics[J]. Small, 2019, 15(39): e1902313.
3	GUO Jingyang, QIN Simin, WEI Yan, et al. Silver nanoparticles exert concentration-dependent influences on biofilm development and architecture[J]. Cell Proliferation, 2019, 52(4): e12616.
4	程峰, 刘宇, 杨珍, 等. 茶树精油对耐甲氧西林金黄色葡萄球菌生物被膜的影响[J]. 食品工业科技, 2021, 42(14): 107-112.
	CHENG Feng, LIU Yu, YANG Zhen, et al. Effect of tea tree oil on methicillin-resistant staphylococcus aureus biofilm[J]. Science and Technology of Food Industry, 2021, 42(14): 107-112.
5	王桂洋. 壳聚糖对希瓦氏菌生物被膜抑制和清除作用的研究[D]. 杭州: 浙江工商大学, 2015.
	WANG Guiyang. Inhibitory and eliminated effect of chitosan on the biofilm of shewanella[D]. Hangzhou: Zhejiang Gongshang University, 2015.
6	GARCIA Lana Glerieide Silva, DE MELO GUEDES Glaucia Morgana, DA SILVA Maria Lucilene Queiroz, et al. Effect of the molecular weight of chitosan on its antifungal activity against Candida spp. in planktonic cells and biofilm[J]. Carbohydrate Polymers, 2018, 195: 662-669.
7	OMIDI Sakineh, KAKANEJADIFARD Ali. Modification of chitosan and chitosan nanoparticle by long chain pyridinium compounds: Synthesis, characterization, antibacterial, and antioxidant activities[J]. Carbohydrate Polymers, 2019, 208: 477-485.
8	EJAZ Sadaf, IHSAN Ayesha, NOOR Tayyaba, et al. Mannose functionalized chitosan nanosystems for enhanced antimicrobial activity against multidrug resistant pathogens[J]. Polymer Testing, 2020, 91: 106814.
9	PHUANGKAEW Tinnakorn, BOORANABUNYAT Nadda, KIATKAMJORNWONG Suda, et al. Amphiphilic quaternized chitosan: Synthesis, characterization, and anti-cariogenic biofilm property[J]. Carbohydrate Polymers, 2022, 277: 118882.
10	PIEGAT Agnieszka, Agata GOSZCZYŃSKA, IDZIK Tomasz, et al. The importance of reaction conditions on the chemical structure of N, O-acylated chitosan derivatives[J]. Molecules, 2019, 24(17): 3047.
11	DOS SANTOS Z M, CARONI A L P F, PEREIRA M R, et al. Determination of deacetylation degree of chitosan: A comparison between conductometric titration and CHN elemental analysis[J]. Carbohydrate Research, 2009, 344(18):2591-2595.
12	车秋凌, 辛梅华, 李明春, 等. O-一氯均三嗪-N, N, N-三甲基壳聚糖的合成及其在羊毛织物抗菌中的应用[J]. 化工进展, 2018, 37(3): 1098-1104.
	CHE Qiuling, XIN Meihua, LI Mingchun, et al. Synthesis and application of O-monochlotriazinyl-N, N, N-trimethyl chitosan for antibacterial wool fabric[J]. Chemical Industry and Engineering Progress, 2018, 37(3): 1098-1104.
13	魏丽杰. 针对壳聚糖季铵盐的衍生化修饰及生物活性研究[D]. 烟台: 中国科学院大学(中国科学院烟台海岸带研究所), 2019.
	WEI Lijie. Preparation and bioactivity of chitosan quaternary ammonium salt[D]. Yantai: Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, 2019.
14	XIA Wei, WEI Xiaoyi, XIE Yuanyuan, et al. A novel chitosan oligosaccharide derivative: Synthesis, antioxidant and antibacterial properties[J]. Carbohydrate Polymers, 2022, 291: 119608.
15	YUE Lin, WANG Min, KHAN Imranmahmood, et al. Preparation and characterization of chitosan oligosaccharide derivatives containing cinnamyl moieties with enhanced antibacterial activities[J]. LWT Food Science and Technology, 2021, 147: 111663.
16	RÚNARSSON Ögmundur Vidar, HOLAPPA Jukka, MALAINER Clemens, et al. Antibacterial activity of N-quaternary chitosan derivatives: Synthesis, characterization and structure activity relationship (SAR) investigations[J]. European Polymer Journal, 2010, 46(6): 1251-1267.
17	LI Zhihan, YANG Fei, YANG Rendang. Synthesis and characterization of chitosan derivatives with dual-antibacterial functional groups[J]. International Journal of Biological Macromolecules, 2015, 75: 378-387.

样品	C/%	N/%	H/%	DD/%	DS/%
CS	41.69	7.75	7.06	86.00	—
TMBC	39.65	6.54	6.78	—	20.10
DMBBC	45.79	7.35	6.94	—	14.68
DMOBC	44.67	6.82	7.78	—	13.14
DMDBC	49.45	7.34	8.18	—	11.29

样品	C/%	N/%	H/%	DD/%	DS/%
CS	41.69	7.75	7.06	86.00	—
TMBC	39.65	6.54	6.78	—	20.10
DMBBC	45.79	7.35	6.94	—	14.68
DMOBC	44.67	6.82	7.78	—	13.14
DMDBC	49.45	7.34	8.18	—	11.29

样品	最低抑菌浓度 /mg·mL^-1		MBIC/MIC	最低杀菌浓度 /mg·mL^-1		MBBC/MBC
样品	MIC	MBIC	MBIC/MIC	MBC	MBBC	MBBC/MBC
CS	1.250	10.000	8	>5.000	>10.000	>2
TMBC	0.313	1.250	4	2.500	10.000	4
DMBBC	0.313	1.250	4	2.500	10.000	4
DMOBC	0.313	1.250	4	2.500	10.000	4
DMDBC	0.156	0.625	4	0.625	1.250	4

样品	最低抑菌浓度 /mg·mL^-1		MBIC/MIC	最低杀菌浓度 /mg·mL^-1		MBBC/MBC
样品	MIC	MBIC	MBIC/MIC	MBC	MBBC	MBBC/MBC
CS	1.250	10.000	8	>5.000	>10.000	>2
TMBC	0.313	1.250	4	2.500	10.000	4
DMBBC	0.313	1.250	4	2.500	10.000	4
DMOBC	0.313	1.250	4	2.500	10.000	4
DMDBC	0.156	0.625	4	0.625	1.250	4

样品	最低抑菌浓度 /mg·mL^-1		MBIC/MIC	最低杀菌浓度 /mg·mL^-1		MBBC/MBC
样品	MIC	MBIC	MBIC/MIC	MBC	MBBC	MBBC/MBC
CS	0.313	2.500	8	1.250	10.000	8
TMBC	0.039	0.313	8	0.313	2.500	8
DMBBC	0.039	0.156	4	0.313	2.500	8
DMOBC	0.039	0.156	4	0.156	2.500	16
DMDBC	0.039	0.156	4	0.156	1.250	8

烷基季铵化壳聚糖的制备及其抗生物被膜的活性

Preparation of alkyl quaternized chitosan and its anti-biofilm activity

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

图/表 11

参考文献 17

相关文章 8

编辑推荐

Metrics

本文评价

[1]	王林, 辛梅华, 李明春, 陈琦, 毛扬帆. 季铵化/磺化壳聚糖的制备及其抗生物被膜活性[J]. 化工进展, 2023, 42(5): 2577-2585.
[2]	张毅, 姜迎雪, 张昊. 季铵型改性壳聚糖的制备及其在兔毛织物抗菌整理中的应用[J]. 化工进展, 2020, 39(7): 2810-2816.
[3]	申向,刘开全,刘鹏,姚思杰,吕妮娜,张永金,汪帆. 辣素衍生物改性PVDF膜的制备及其抗菌性能[J]. 化工进展, 2019, 38(12): 5427-5434.
[4]	车秋凌, 辛梅华, 李明春, 陈帅. O-一氯均三嗪-N,N,N-三甲基壳聚糖的合成及其在羊毛织物抗菌中的应用[J]. 化工进展, 2018, 37(03): 1098-1104.
[5]	邱松山, 姜翠翠, 周如金, 刘杰凤. 反丁烯二酸酯类物质结构与抑菌性能关系[J]. 化工进展, 2016, 35(03): 879-883.
[6]	陈燕燕, 李明春, 辛梅华, 林意华. 海因改性N-季铵化壳聚糖衍生物的合成及其抗菌性能[J]. 化工进展, 2015, 34(1): 188-192,233.
[7]	周盛全，辛梅华，李明春，宋宇宁，王朝. 新型壳聚糖季铵盐抗菌剂的合成及其性能[J]. 化工进展, 2012, 31(08): 1801-1805.
[8]	李明春，许涛，辛梅华. 壳聚糖及其衍生物的抗菌活性研究进展 [J]. 化工进展, 2011, 30(1): 203-.