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CN105621627B - 一种利用TiO2‑CeO2负载CYP119酶光催化还原Cr6+的方法 - Google Patents

一种利用TiO2‑CeO2负载CYP119酶光催化还原Cr6+的方法 Download PDF

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CN105621627B
CN105621627B CN201510972547.2A CN201510972547A CN105621627B CN 105621627 B CN105621627 B CN 105621627B CN 201510972547 A CN201510972547 A CN 201510972547A CN 105621627 B CN105621627 B CN 105621627B
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enzyme
cyp119
ethanol
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CN105621627A (zh
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蒋华麟
陈萍华
厉梦琳
刘军
田磊
张为波
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Nanchang Hangkong University
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    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00—Biological treatment of water, waste water, or sewage
    • C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
    • C02F3/342—Biological treatment of water, waste water, or sewage characterised by the microorganisms used characterised by the enzymes used
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Abstract

本发明涉及一种利用TiO2‑CeO2负载CYP119酶光催化还原Cr6+的方法，所得材料能在光照下高效率地催化还原重金属环境污染物Cr6+。所述方法包括：将Ce(NO3)3·6H2O和钛酸正丁酯溶液混和均匀后，转至水热釜反应。反应结束后将样品分离，清洗，烘干后煅烧得到TiO2‑CeO2。以TiO2‑CeO2为载体负载CYP119酶制备得到光催化剂。其特征是利用TiO2‑CeO2负载CYP119酶，所得材料具有高效的光催化还原Cr6+的性能。

Description

一种利用TiO2-CeO2负载CYP119酶光催化还原Cr6+的方法
技术领域
本发明涉及光催化及生物催化技术领域，尤其涉及一种利用TiO2-CeO2负载CYP119酶光催化还原Cr6+的方法。
背景技术
CYP119酶是一种细胞色素P450酶，具有超常的稳定性和耐受性。它的热变性温度高达90℃，并可以耐受200 MPa以上的高压和极端的pH条件，是极佳的生物催化剂，是蛋白质工程的理想目标，在生物制药、生物合成等领域具有极其重要的应用价值。最近的研究显示其具有特别的光激活机制，这为构建基于CYP119酶的光催化体系提供了依据，使利用简单廉价的光催化体系代替复杂昂贵的辅助酶体系提供了可能，这将极大地推动酶催化体系在环境污染物治理中的实际应用。
六价铬化合物在皮革、化工、电子产品中广泛应用，残留在环境中的Cr6+是一种严重的环境污染物。Cr6+在体内具有致癌作用，还会引起诸多的其他健康问题，如吸入某些较高浓度的六价铬化合物会引起流鼻涕、打喷嚏、瘙痒、鼻出血、溃疡和鼻中隔穿孔。短期大剂量的接触，在接触部位会产生不良后果，包括溃疡、鼻黏膜刺激和鼻中隔穿孔。摄入超大剂量的铬会导致肾脏和肝脏的损伤、恶心、胃肠道刺激、胃溃疡、痉挛甚至死亡。危害最大的是长期或短期接触或吸入时有致癌危险。因此，治理Cr6+污染意义重大。
在本发明方法中，提供了一种构建基于CYP119酶光催化体系的方法。利用TiO2-CeO2复合材料作为载体，负载CYP119酶，构建复合光催化材料。所得产品对Cr6+有很好的光催化还原效果。
本发明涉及一种利用TiO2-CeO2负载CYP119酶光催化还原Cr6+的方法，原料易得，成本低，操作简单，所述方法未见报道。
发明内容
本发明的目的在于提供一种利用TiO2-CeO2负载CYP119酶光催化还原Cr6+的方法，本发明采取如下手段：
一种利用TiO2-CeO2负载CYP119酶光催化还原Cr6+的方法，其特征在于：
（1）将Ce(NO3)3·6H2O溶解于乙醇，比例为0.1498 g Ce(NO3)3·6H2O/20 mL乙醇；
（2）将钛酸正丁酯溶解于乙醇和H2O的混和液，比例为352 ul钛酸正丁酯/20 mL乙醇和10 mL H2O的混和液；
（3）将（1）和（2）所得溶液混和，超声分散均匀，转入水热反应釜120℃水热反应48h。将所得固体离心分离，干燥后450℃煅烧5 h，得到TiO2-CeO2复合材料；
（4）将CYP119酶溶液与TiO2-CeO2复合材料通过持续搅拌混和分散均匀，然后离心分离，洗涤后冷冻干燥即得产品。
所得的材料利用TiO2-CeO2复合材料负载CYP119酶，构建酶光催化体系。
所得的材料具有良好的光催化还原活性，对Cr6+具有很好的光催化还原性能。
本发明的优点是：利用TiO2-CeO2作为载体负载CYP119酶，构建新型的酶光催化体系，使得CYP119酶的光活性能够得到很好的应用，对推动酶催化体系在环境污染物治理中的实际应用具有重要意义；原料易得，成本低，操作简单；所得产品具有优良的光催化还原性能，可利用光催化还原重金属环境污染物Cr6+。
附图说明
图1为本发明之实施例1所得到TiO2-CeO2的扫描电镜图；
图2为本发明之实施例1所得到材料光催化还原Cr6+。
具体实施方式
实施例1
不断搅拌下将0.1498 g Ce(NO3)3·6H2O溶解于20 mL乙醇；在20 mL乙醇和10 mLH2O混和液中加入352 ul钛酸正丁酯；将上述两种溶液混和，通过超声分散均匀，再转入水热反应釜于120℃水热反应48 h后，将所得固体离心分离，干燥后于450℃煅烧5 h，得到TiO2-CeO2。将50 mg TiO2-CeO2分散于50 mL pH=6.5的磷酸钾缓冲溶液中，再加入200 uL的1 uM的CYP119酶溶液，持续搅拌20 min，离心分离得到固体，洗涤后冷冻干燥，即得到产品。
将所得产品用于降解初始浓度为2 mM Cr6+溶液，考查产品的光催化性能。
实施例2
不断搅拌下将0.2996 g Ce(NO3)3·6H2O溶解于40 mL乙醇；在40 mL乙醇和20 mLH2O混和液中加入704 ul钛酸正丁酯；将上述两种溶液混和，通过超声分散均匀，再转入水热反应釜于120℃水热反应48 h后，将所得固体离心分离，干燥后于450℃煅烧5 h，得到TiO2-CeO2。将50 mg TiO2-CeO2分散于50 mL pH=6.5的磷酸钾缓冲溶液中，再加入200 uL的1 uM的CYP119酶溶液，持续搅拌20 min，离心分离得到固体，洗涤后冷冻干燥，即得到产品。
将所得产品用于降解初始浓度为2 mM Cr6+溶液，考查产品的光催化性能。

Claims (2)

1.一种利用TiO2-CeO2负载CYP119酶光催化还原Cr6+的方法，其特征在于：
（1）将Ce(NO3)3·6H2O溶解于乙醇，比例为0.1498 g Ce(NO3)3·6H2O/20 mL乙醇；
（2）将钛酸正丁酯溶解于乙醇和H2O的混和液，比例为352 μL 钛酸正丁酯/20 mL乙醇和10 mL H2O的混和液；
（3）将（1）和（2）所得溶液混和，超声分散均匀，转入水热反应釜120℃水热反应48 h；将所得固体离心分离，干燥后450℃煅烧5 h，得到TiO2-CeO2复合材料；
（4）将CYP119酶溶液与TiO2-CeO2复合材料通过持续搅拌混和分散均匀，然后离心分离，洗涤后冷冻干燥即得产品；
所得的产品具有良好的光催化还原活性，对Cr6+具有很好的光催化还原性能。
2.根据权利要求1所述一种利用TiO2-CeO2负载CYP119酶光催化还原Cr6+的方法，其特征在于：所得的产品利用TiO2-CeO2复合材料负载CYP119酶，构建酶光催化体系。
CN201510972547.2A 2015-12-23 2015-12-23 一种利用TiO2‑CeO2负载CYP119酶光催化还原Cr6+的方法 Expired - Fee Related CN105621627B (zh)

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CN109317095A (zh) * 2018-10-11 2019-02-12 温州医科大学 一种用于六价铬去除的复合材料及其制备方法
CN111167519B (zh) * 2020-01-13 2022-08-05 河北工业大学 光酶集成纳米催化剂及其应用

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CN101775387A (zh) * 2010-03-23 2010-07-14 南京工业大学 一种介孔二氧化钛固定化酶及其制备方法和应用
KR20100125845A (ko) * 2009-05-22 2010-12-01 전남대학교산학협력단 광촉매를 이용한 리그노셀룰로스계 바이오매스의 전처리 방법 및 이를 이용한 바이오에탄올의 생산 방법
RU2416644C1 (ru) * 2009-11-17 2011-04-20 Российская Федерация, От Имени Которой Выступает Министерство Образования И Науки Российской Федерации Фотобиокатализатор для получения восстановленных форм никотинамидных коферментов nadh или nadph и фотокаталитический способ получения nadh или nadph

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1796552A (zh) * 2004-12-29 2006-07-05 华东理工大学 一种用于生物酶载体的块状多孔材料的制备方法
KR20100125845A (ko) * 2009-05-22 2010-12-01 전남대학교산학협력단 광촉매를 이용한 리그노셀룰로스계 바이오매스의 전처리 방법 및 이를 이용한 바이오에탄올의 생산 방법
RU2416644C1 (ru) * 2009-11-17 2011-04-20 Российская Федерация, От Имени Которой Выступает Министерство Образования И Науки Российской Федерации Фотобиокатализатор для получения восстановленных форм никотинамидных коферментов nadh или nadph и фотокаталитический способ получения nadh или nadph
CN101775387A (zh) * 2010-03-23 2010-07-14 南京工业大学 一种介孔二氧化钛固定化酶及其制备方法和应用

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