【摘要】：本文以染料活性艷藍(Remazol Brilliant Blue KN-R, RBBR)模擬染料廢水,利用過水解酶原位催化制備過氧乙酸的方法對其進行脫色處理,并對脫色條件進行了優(yōu)化。同時,采用交聯(lián)酶聚體技術(shù)對過水解酶進行固定化,并用聚乙烯醇進行二次包埋。最后利用固定化酶進行染料RBBR脫色。主要的實驗結(jié)果如下：1、過水解酶的純化及酶學(xué)性質(zhì)分析。利用親和層析柱純化過水解酶,SDS-PAGE電泳顯示為單一條帶,其分子量大小為28 kDa,比活力為9.03 U/mg。酶學(xué)性質(zhì)分析表明,該過水解酶最適pH為6.0,最適溫度為65℃,在pH 3.0-6.0及溫度為30-65℃范圍內(nèi)具有較好的穩(wěn)定性；其T502h為83.4℃。2、過水解酶催化的脫色反應(yīng)。利用過水解酶原位催化制備過氧乙酸對染料RBBR進行脫色。在單因子試驗的基礎(chǔ)上,通過正交優(yōu)化試驗獲得最優(yōu)脫色條件：在5 mL反應(yīng)體系中,最適pH5.0、酶量20 U/Reaction、乙酸乙酯與過氧化氫的摩爾比40：1、染料濃度80 mg/mL。在最優(yōu)條件下處理6h后,染料RBBR的脫色率為81.11%,24 h的脫色率為91.96%。將體系放大50倍后(250 mL),24 h染料RBBR的脫色率為84.55%。3、過水解酶的固定化及應(yīng)用效果評估。首先采用交聯(lián)酶聚集體技術(shù)(CLEAs)對過水解酶進行固定化,然后采用聚乙烯醇對交聯(lián)過水解酶聚集體進行二次包埋,進行脫色反應(yīng)。結(jié)果表明,當(dāng)使用60%(v/v)的有機溶劑S2為沉淀劑,且酶與BSA的比率為16：2,交聯(lián)劑戊二醛的濃度為100mmol/L時,制備得到的過水解酶交聯(lián)酶聚體(Per-CLEAs)酶活回收率可達79.72%。利用不同濃度的聚乙烯醇(PVA)進行二次包埋固定化,結(jié)果發(fā)現(xiàn),當(dāng)PVA濃度為10%(w/v)時,包埋后的Per-CLEAs形態(tài)較好、酶活回收率較高,而且洗滌三次后無明顯的酶活泄露。用其對染料RBBR脫色,在維持80%的脫色率的前提下,至少可以進行6次循環(huán)。分別利用游離酶、Per-CLEAs以及包埋后的Per-CLEAs催化合成過氧乙酸原位氧化RBBR脫色,處理30 min后,脫色率分別為63.72%、95.98%、83.51%。
[Abstract]:In this paper, dye reactive brilliant blue (Remazol Brilliant Blue KN-R, RBBR (reactive brilliant blue dye) was used to simulate dye wastewater, and the decolorization conditions were optimized by in-situ catalytic preparation of peracetic acid by hyperhydrolysis enzyme. At the same time, cross-linked enzyme polymer technology was used to immobilize the hydrolase, and polyvinyl alcohol was used to encapsulate the hydrolase for two times. Finally, the dye RBBR was decolorized by immobilized enzyme. The main results are as follows: 1. Purification and characterization of hydrolase. The hydrolase was purified by affinity chromatography. SDS-PAGE electrophoresis showed that the hydrolase was a single band with a molecular weight of 28 kDa, and a specific activity of 9.03 U ~ (?) mg 路g ~ (- 1). The results showed that the optimal pH and temperature of the hydrolase were 6.0 鈩，

本文編號：2451315