兩種微球載體固定化脂肪酶的研究
本文選題:脂肪酶 + 微球。 參考:《河北大學(xué)》2014年碩士論文
【摘要】:脂肪酶在實(shí)際應(yīng)用中的催化能力具有高效性,在反應(yīng)過(guò)程中副反應(yīng)和副產(chǎn)物比非生物催化劑少,且在較溫和的環(huán)境下就能進(jìn)行催化反應(yīng),因此,被廣泛地應(yīng)用到了工業(yè)化生產(chǎn)中。然而其穩(wěn)定性差和難回收等缺點(diǎn)限制了其在生產(chǎn)中的應(yīng)用,為了克服這些缺點(diǎn),將游離酶通過(guò)物理吸附法、包埋法或共價(jià)結(jié)合法進(jìn)行固定化研究。 本文分為兩部分。第一部分,首先以苯乙烯(St)和丙烯酸(AA)為單體,通過(guò)乳液聚合法,制備出具有羧基功能基團(tuán)的P(St-AA)磁性微球,再以所制備的磁性微球?yàn)檩d體,通過(guò)共價(jià)結(jié)合的方法對(duì)Candida rugosa lipase進(jìn)行固定化,,考察了不同長(zhǎng)度的間隔臂鏈(乙二胺和PEG400/800/4000)修飾到磁性微球上對(duì)克服空間位阻的影響。結(jié)果表明,當(dāng)使用PEG800/4000作為間隔臂鏈時(shí),可以有效地克服空間位阻。結(jié)果還表明,過(guò)高的酶載量會(huì)產(chǎn)生空間位阻而影響酶的活性。并進(jìn)一步研究了PEG800在磁性微球上的修飾量對(duì)固定化酶活性的影響。 第二部分,以乙酸乙烯酯(VAC)、丙烯酰胺(AM)為單體,通過(guò)懸浮聚合法制備了P(VAC-AM)高分子微球,并以此高分子微球?yàn)檩d體,通過(guò)環(huán)氧氯丙烷共價(jià)結(jié)合Candidarugosa lipase,考察了高分子微球環(huán)氧基的修飾條件和脂肪酶的固定化條件。由結(jié)果可以看出,轉(zhuǎn)速為600r/min,環(huán)氧氯丙烷的用量為3mL,酶的用量為5mg,并且固定化時(shí)間為2h時(shí),固定化酶的活性回收率達(dá)到最大為48%。
[Abstract]:The catalytic ability of lipase in practical application is effective, the side reaction in the process and the by-product of non biological catalyst, and can be carried out under mild reaction conditions, therefore, it is widely applied in industrial production. However, its poor stability and difficult recovery in their application is limited. In the production, in order to overcome these shortcomings, the free enzyme by physical adsorption or covalent binding method of embedding immobilization.
This paper is divided into two parts. The first part, firstly, using styrene (St) and acrylic acid (AA) as the monomer by emulsion polymerization, prepared with carboxyl functional groups of P (St-AA) magnetic microspheres, magnetic microspheres with the prepared as the carrier, by covalent coupling method for immobilization Candida rugosa lipase, investigated the spacer chains of different lengths (ethylenediamine and PEG400/800/4000) modified magnetic microspheres to overcome the steric effects. The results show that when using PEG800/4000 as the spacer chain, can effectively overcome the steric hindrance. The results also showed that high enzyme loading produces influence enzyme the activity of steric hindrance. And further study the effects of modified PEG800 on the magnetic microspheres on the activity of immobilized enzyme.
The second part, with vinyl acetate (VAC), acrylamide (AM) monomers were prepared by suspension polymerization of P (VAC-AM) microspheres, and the polymer microspheres by epoxy chloropropane covalently bound Candidarugosa lipase immobilized conditions investigated polymer microspheres epoxy modification conditions and lipase. The test results show that the speed of 600r/min, epichlorohydrin dosage is 3mL, the dosage of enzyme is 5mg, and the immobilization time of 2h, the activity recovery rate of immobilized enzyme reached 48%.
【學(xué)位授予單位】:河北大學(xué)
【學(xué)位級(jí)別】:碩士
【學(xué)位授予年份】:2014
【分類(lèi)號(hào)】:R917
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