本文關(guān)鍵詞： 殼聚糖 胍基乙酸化殼聚糖 金黃色葡萄球菌 大腸桿菌 抗菌性能　出處：《華中科技大學(xué)》2012年碩士論文　論文類(lèi)型：學(xué)位論文

【摘要】：殼聚糖(Chitosan,CS)是自然界中唯一已知的含氮堿性多糖，有良好的生物相容性、可降解性、抗菌性。殼聚糖及其衍生物在促進(jìn)傷口愈合、抗菌和抗病毒等領(lǐng)域的應(yīng)用日益引人注目。殼聚糖衍生物抗菌性能隨其所帶正電性增強(qiáng)而增大,胍基是目前發(fā)現(xiàn)的自然界正電性最強(qiáng)的有機(jī)基團(tuán)，引入胍基基團(tuán)能增強(qiáng)殼聚糖的抗菌性能。因而本文設(shè)計(jì)了一種抗菌性胍基化殼聚糖即胍基乙酸化殼聚糖（CS-N-Guanidine，CG），并測(cè)定殼聚糖及其胍基乙酸化殼聚糖的抗菌性能并得出相關(guān)結(jié)論。 本文通過(guò)對(duì)甲殼素進(jìn)行脫乙酰得到脫乙酰度較低的殼聚糖，再進(jìn)行純化、脫乙酰、分子量降解之后得到脫乙酰度為83.15%，分子量為80.4kDa的純化殼聚糖。以EDC(1-(3-二甲氨基丙基)-3-乙基碳二亞胺)為引發(fā)劑，NHS(N-羥基硫代琥珀酰亞胺)為催化劑，MES(2-(N-嗎啉)乙磺酸)為緩沖液將胍基乙酸接枝到純化殼聚糖的自由氨基上得到胍基乙酸化殼聚糖，控制反應(yīng)時(shí)間18h，24h，36h得到三種取代度分別為11.03%、13.39%、21.59%的胍基乙酸化殼聚糖。采用瓊脂糖平板計(jì)數(shù)法測(cè)定胍基乙酸化殼聚糖對(duì)革蘭氏陽(yáng)性菌金黃色葡萄球菌（Staphylococcus aureus, S. aureus）和革蘭氏陰性菌大腸桿菌（Escherichia coli，E. coli）的抑制作用。 結(jié)果表明：CS和CG對(duì)金黃色葡萄球菌和大腸桿菌有抑制作用，且CG的抑制作用比CS明顯，CG的抗菌性能隨取代度的提高而增強(qiáng)。 CS在濃度＜0.3g/L時(shí)對(duì)金黃色葡萄球菌有微弱的促進(jìn)作用，在濃度＞0.3g/L能夠完全抑制金黃色葡萄球菌的生長(zhǎng)；在濃度＜0.2g/L對(duì)大腸桿菌有微弱的促進(jìn)作用，隨著濃度的提高抑菌效果越來(lái)越明顯，在濃度＞0.3g/L時(shí)能完全抑制大腸桿菌的生長(zhǎng)。 CG在濃度＜0.15g/L時(shí)能夠促進(jìn)金黃色葡萄球菌的生長(zhǎng)，，隨濃度的提高抑菌性能也得到提高，且取代度越高，抑菌效果越明顯，在濃度＞0.15g/L時(shí)金黃色葡萄球菌的生長(zhǎng)完全受到抑制；CG在濃度＜0.1g/L時(shí)能促進(jìn)大腸桿菌的生長(zhǎng)，濃度＞0.1g/L時(shí)表現(xiàn)出抑制作用，且取代度越高抑制效果越明顯，當(dāng)濃度達(dá)到0.15g/L時(shí)可以完全抑制大腸桿菌的生長(zhǎng)。
[Abstract]:Chitosan Chitosanine (CSS) is the only nitrogen-containing alkaline polysaccharide known in nature with good biocompatibility biodegradability and antibacterial properties. Chitosan and its derivatives promote wound healing. Antimicrobial and antiviral applications have become increasingly attractive. The antibacterial properties of chitosan derivatives increase with the increase of positive charge. Guanidine is the most active organic group found in nature. The introduction of guanidine groups can enhance the antibacterial properties of chitosan. Therefore, a kind of antimicrobial guanidine chitosan (CS-N-Guanidine CGG) was designed. The antimicrobial properties of chitosan and its guanidine acetate chitosan were determined and the relevant conclusions were obtained. In this paper, chitosan with low degree of deacetylation was obtained by deacetylation of chitin, then purified, deacetylated, and deacetylated to 83.15% after molecular weight degradation. A purified chitosan with molecular weight of 80.4 kDa was prepared by using EDCA (1-dimethylaminopropyl) -3-ethylcarbodiimide as initiator. NHSN-hydroxythiosuccinimide) was used as catalyst. The guanidinoacetic acid was grafted onto the free amino group of purified chitosan, and the reaction time was 18 h / 24 h. The degree of substitution was 11.03% and 13.39% respectively at 36h. 21.59% guanidine acetate chitosan. Agarose plate counting method was used to determine the effect of guanidine acetate chitosan on Gram-positive bacteria Staphylococcus aureus (Staphylococcus aureus). Staphylococcus aureus. The inhibitory effects of Saureus and Escherichia coli E. coli. The results showed that the inhibitory effect of CG on Staphylococcus aureus and Escherichia coli was more obvious than that of CS, and the antibacterial activity of CG increased with the increase of substitution degree. CS had a slight effect on Staphylococcus aureus when the concentration was less than 0.3 g / L, and could completely inhibit the growth of Staphylococcus aureus when the concentration was more than 0.3 g / L. The bacteriostatic effect of Escherichia coli was more and more obvious when the concentration was less than 0.2 g / L, and the growth of Escherichia coli could be completely inhibited when the concentration was more than 0.3 g / L. CG could promote the growth of Staphylococcus aureus when the concentration of CG was less than 0.15 g / L, and the higher the degree of substitution, the more obvious the bacteriostatic effect was. The growth of Staphylococcus aureus was completely inhibited when the concentration was more than 0.15 g / L. CG could promote the growth of Escherichia coli when the concentration was less than 0.1 g / L, and when the concentration was higher than 0.1 g / L, the inhibitory effect was more obvious, and the higher the degree of substitution was, the more obvious the inhibitory effect was. When the concentration reached 0.15 g / L, the growth of Escherichia coli was completely inhibited.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類(lèi)號(hào)】：R318.08

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