CN110813263A - 固定化酵母的生物质基水凝胶吸附剂及其制备方法和应用 - Google Patents
固定化酵母的生物质基水凝胶吸附剂及其制备方法和应用 Download PDFInfo
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Abstract
本发明公开了一种固定化酵母的生物质基水凝胶及其制备方法,本发明所述的水凝胶具有三维大孔网络结构,吸附位点充分暴露,水溶性小分子可自由扩散,而对水溶液中金属离子具有很高的吸附效率的双网络水凝胶吸附剂。本发明将具有吸附效果的微生物固定到凝胶吸附剂中,其对水溶性小分子物质可充分扩散,而对水中重金属离子吸附效果优异,且环境友好,成本低,便于推广使用。
Description
技术领域
本发明涉及涉及一种固定化酵母的生物质基凝胶的制备及其作为重金属吸附剂的应用,属于环境功能性材料和水污染处理技术领域。
背景技术
由于水体中重金属污染的环境持久性、非生物降解性、易生物积累性和致死毒性,即使在相对低的浓度下对人类和生物体具有严重的毒性作用。近年来,多种处理技术被用于去除废水中的有毒金属离子污染物,如离子交换、过滤、化学沉淀、反渗透、氧化还原、电凝聚、吸附等。与其他方法相比吸附法具有效率高、成本低、易于操作和实施以及不会造成二次污染等优点。因此制备出一种低成本、高效率、易分离的吸附材料对控制废水中重金属含量具有重要意义。
水凝胶是一种重要的功能高分子材料,广泛应用于医药和软机械领域。作为吸附材料,它具有残留少、吸附效率高、可重复使用等优点,是一类很有前景的重金属吸附剂,但传统水凝胶由于缺乏有效的能量耗散机制,强度和韧性不足,限制了其应用。而双网络水凝胶因其可控的性质和优异的机械性能促使我们通过引入官能团来开发这种材料作为吸附剂。目前,已成功制备了部分双网络水凝胶材料应用于吸附重金属离子,如壳聚糖/聚丙烯酰胺凝胶(Ma et al. ,2016. Efficient removal of heavy metal ions with an EDTAfunctionalized chitosan/polyacrylamide double network hydrogel. ACSSustainable Chemistry & Engineering. 5(1): 843-851.),但是需要在制备出来凝胶之后进行功能性修饰才能达到理想的吸附效果。
由于我国茶叶需求量大,每年都会产生大量的茶渣,可作为理想的低成本生物质资源。茶渣资源高效利用的主要途径是制备活性炭吸附剂,但活性炭制备成本高,限制了其推广利用。茶渣主要由纤维素、半纤维素、木质素、缩合单宁和蛋白组成,富含羧酸盐、芳香羧酸、酚羟基和氧基,它们可以与重金属离子螯合或配位。利用微生物的吸收和吸附作用脱除重金属的方法叫做微生物法,研究显示,一些酵母菌,细菌,霉菌等具有优异的脱除重金属效果,但是微生物法存在难以分离回收的技术难题。因此,将具有吸附效果的微生物与生物质充分结合制备出一种重金属吸附剂,既可解决上述难题,还能大幅度提高材料的吸附效果。目前,还没有关于固定化酵母的生物质基高性能双网络水凝胶吸附剂的有关报道。
发明内容
本发明要解决的技术问题是提供一种生物基质耦合微生物制备水凝胶来吸附重金属,优选的是利用废弃茶渣生物质资源,耦合酵母细胞来制备力学性能优良,富含活性基团,具有交联纤维素及聚丙烯酸的大孔双网络三维水凝胶及其制备方法,该方法操作简单,成本低,利于大规模生产。
生物基质耦合微生物可以制备水凝胶,生物基质可以选择茶渣、咖啡渣、玉米秸秆、柚子皮、花生壳、草灰、木炭;
微生物包括酵母菌、细菌、霉菌。具体的:
酵母菌包括库德毕赤酵母、酿酒酵母、圆红冬孢酵母菌。
细菌包括伯克氏菌、蜡样芽孢杆菌、植物乳杆菌、鞘细菌。
霉菌包括黑曲霉菌、拟青霉菌、毛霉菌、卡伍尔链霉菌、荧光假单胞菌。
本发明提供了一种水凝胶,包括茶渣和酵母细胞。所述的水凝胶的制备方法,包括下列步骤:
(一) 制备酵母菌泥制备。
(二) 茶渣粉水溶液制备。
(三)茶渣水凝胶的制备与处理。
步骤(一)制备酵母菌泥制备的具体步骤为:
将菌种接种到斜面培养基上,在恒温培养箱中培养24h后取出,接种到YPD中,在摇床上扩大培养24h;然后取1mL菌液,分别加入YPD及用NaCl胁迫的YPD中,在摇床上培养24h后,将菌液分别置于5mL的离心管中,3700r离心5min,即得到菌泥,菌泥中菌的数量为3-5×106。
步骤(二)茶渣粉水溶液制备的具体步骤为:
分别配置0.5 mol/L的氢氧化钠溶液和氯化氢溶液,量取0.5 mol/L 的10.5 mL的氢氧化钠溶液,将茶渣粉过100目筛,加入1-5g茶渣粉,混合搅拌1-30 min后加入0.5 mol/L 的10.5 mL盐酸溶液调至中性,得到茶渣水溶液。
步骤(三)茶渣水凝胶的制备与处理的具体步骤为:
茶渣水溶液中加入0.3 g过硫酸铵,0.1215 g N,N-亚甲基双丙烯酰胺,3.6 mL丙烯酸和5 mL菌液,菌液为菌泥加去离子水重新稀释至5mL变成菌液;
搅拌混匀后,分装到模具中,每个模具中含有5 mL混合物,充入氮气密封,在60℃条件下水浴3-8 h;水浴结束后,形成硬度、强度较好的包埋酵母菌的茶渣水凝胶;
取出反应结束的水凝胶,于去离子水中充分透析24h,每隔2-6 h换一次水;透析结束后,取出水凝胶冻干,备用。
本发明提供的水凝胶可以用在吸附重金属上。
本发明的有益效果:
本发明所述的水凝胶具有三维大孔网络结构,吸附位点充分暴露,水溶性小分子可自由扩散,而对水溶液中金属离子具有很高的吸附效率的双网络水凝胶吸附剂。
本发明将具有吸附效果的微生物固定到凝胶吸附剂中,其对水溶性小分子物质可充分扩散,而对水中重金属离子吸附效果优异,且环境友好,成本低,便于推广使用。
附图说明
图1是扫描电镜结果图。
图2是力学性能测试图。
图3是溶胀性能测试图。
图4是吸附性能图。
图5是循环性能图。
具体实施方式
实施例1:
本实施例提供一种水凝胶,包括茶渣和库德毕赤酵母细胞。
制备方法包括下列步骤:
(一) 制备库德毕赤酵母菌泥制备。
(二) 茶渣粉水溶液制备。
(三)茶渣水凝胶的制备与处理。
步骤(一)制备酵母菌泥制备的具体步骤为:
将菌种接种到斜面培养基上,在恒温培养箱中培养24h后取出,接种到YPD中,在摇床上扩大培养24h;然后取1mL菌液,分别加入YPD及用NaCl胁迫的YPD中,在摇床上培养24h后,将菌液分别置于5mL的离心管中,3700r离心5min,即得到菌泥,菌泥中菌的数量为4×106。
步骤(二)茶渣粉水溶液制备的具体步骤为:
分别配置0.5 mol/L的氢氧化钠溶液和氯化氢溶液,量取0.5 mol/L 的10.5 mL的氢氧化钠溶液,将茶渣粉过100目筛,加入2g茶渣粉,混合搅拌10 min后加入0.5 mol/L 的10.5mL盐酸溶液调至中性,得到茶渣水溶液。
步骤(三)茶渣水凝胶的制备与处理的具体步骤为:
茶渣水溶液中加入0.3 g过硫酸铵,0.1215 g N,N-亚甲基双丙烯酰胺,3.6 mL丙烯酸和5 mL菌液,菌液为菌泥加去离子水重新稀释至5mL变成菌液;
搅拌混匀后,分装到模具中,每个模具中含有5 mL混合物,充入氮气密封,在60℃条件下水浴6h;水浴结束后,形成硬度、强度较好的包埋酵母菌的茶渣水凝胶;
取出反应结束的水凝胶,于去离子水中充分透析24h,每隔4h换一次水;透析结束后,取出水凝胶冻干,备用。
实施例2:实施例1的水凝胶的验证
1 扫描电镜结果
取少量经冷冻干燥后的未研磨TR/PAA水凝胶样品,用导电胶分别固定于铜板上,对样品表面进行喷金处理,用VEGA3扫描电子显微镜分别在20kV加速电压下观察样品的表面及横断面形貌,来确定其微观结构。水凝胶溶胀使其内部结构中含有大量水分,将其冷冻干燥处理,扫描电镜(SEM)下研究了溶胀后TR/PAA干凝胶的微观结构和表面形貌(如图1-d)可以清楚地看出TR/PAA 干凝胶内部呈三维多孔网络结构,内部具有彼此相通且不规则的孔道,根据图上标尺可以估测孔径范围在100.0 ~ 200.0 μm 之间,属于大孔结构。显然,这种松散、多孔结构吸附剂中的官能团能够完全暴露于金属离子,且增加了水凝胶的比表面积,有益于外界溶液中的重金属离子快速扩散到达凝胶内部的吸附位点,从而增加了凝胶内部对重金属的吸附容量。此外,块状结构的 TR/PAA 水凝胶便于重复使用,有利于从水中快速分离,且可根据实际需要将凝胶切成合适的尺寸。
2.力学性能测试
室温下,将圆柱形的湿凝胶样品置于Z020型万能材料试验机的下板上,通过测力传感器连接的上板以 2.0 mm·min-1的压缩速度压缩,进行形变量分别为30%、50%和70%的循坏加载-卸载测试,三次循坏后,接着测试直至该凝胶压出现不可逆性的破碎,分析此时该凝胶所能承受的最大压力。根据结果可知,凝胶可承受高达350 kPa的压力,说明该材料力学性能优良。
3. 溶胀性能测试
在蒸馏水中通过重量法确定水凝胶的溶胀比。将干燥的完整水凝胶样品在室温(298K)下浸入到过量的蒸馏水中。在预设的时间间隔后,从水介质中取出溶胀的水凝胶,并使用滤纸轻轻拭去表面多余的水之后立即称重,监测直到恒重。每个样品称重3次,取平均值。水凝胶样品的平衡溶胀比(SR)由以下公式确定:
SR(%)=(We-Wd)/ Wd×100%
式中,We是水凝胶的湿重,Wd是水凝胶的干重。
在初始阶段,由于水凝胶结构中存在的大量阴离子基团增大了材料的渗透压,此时水分子受到的阻力相对较小,故反应初期具有较高的溶胀速率,随着溶胀过程进行,凝胶内部充满越多水分子,渗透压差逐渐减弱,水分子需要克服越来越大的渗透压,溶胀速率减慢,约7740min后水凝胶达到最大溶胀,此时溶胀比高达15.9,且凝胶仍然能保持原有形状,说明该凝胶稳定性良好。经充分溶胀后的水凝胶含水量高达94.14%,而高孔隙率和高透水性可以为金属离子从溶液中扩散到水凝胶中提供一条快速通道。
4.吸附性能
预先配制1000 mg·mL-1重金属储备液,实验所用的重金属溶液均用储备液梯度稀释,并通过滴加稀 HNO3或NaOH 溶液来调节整个实验溶液的初始 pH值。设定条件参数为:温度为298 K;每次取10 mL重金属液体于10 mL离心管中;吸附剂的量为0.01 g;摇床转速:160rpm;时间为24 h。为了确保重金属离子在吸附测试中不发生沉淀,对于Pb(Ⅱ), Cr(Ⅲ),Zn(Ⅱ), Cu(Ⅱ), As(Ⅴ), Cd(Ⅱ), Ni(Ⅱ)和Mn(Ⅱ)将pH设定为5 ± 0.01,Fe(III)的pH设定为2.5 ± 0.01。测试Cr(III)、Pb(II)和Fe(III)在凝胶上的竞争吸附时,设定混合溶液中各离子浓度相同,各离子总浓度为1.5 mmol-1。
实验结果表明,对Zn(Ⅱ), Cu(Ⅱ), As(Ⅴ), Cd(Ⅱ), Ni(Ⅱ)和Mn(Ⅱ)的去除能力略差,对Pb(Ⅱ), Cr(Ⅲ),Fe(III)的去除能力较好。对Cr(Ⅲ)的去除率高达98.52%,对Pb(Ⅱ)和Fe(Ⅲ)的去除率分别为90.51%和89.49%。在单组分体系中,凝胶对Cr(III)、Pb(II)和Fe(III)的吸附量分别为226.2、273.2和104.9mg g-1。在三元体系中,凝胶表现出Fe(III)> Cr(III)> Pb(II)的吸附顺序。综上说明所制备的凝胶材料对Cr(III)、Pb(II)和Fe(III)三种金属离子具有优异的吸附能力。
5.循环性能
为了探讨洗脱剂(HCl或EDTA)对金属离子洗脱性能的影响,采用不同浓度的HCl和EDTA对金属离子进行了洗脱,如图所示,随着HCl或EDTA浓度的增加,解吸能力增强,而HCl的解吸效果高于相同浓度的EDTA,对Cr(III)、Pb(II)和Fe(III)解吸最大值分别为94.8%,92.8%和95.8%。为了保证完全解吸,将解吸时间延长到4h,连续7次吸附-解吸后,TR/PAA凝胶对Cr(III)、Fe(III)和Pb(II)的吸附量分别下降了9.9%、5.0%和16.7%。说明凝胶具有良好的再生性能。
当然,上述说明并非是对本发明的限制,本发明也并不限于上述举例,本技术领域的普通技术人员,在本发明的实质范围内,作出的变化、改型、添加或替换,都应属于本发明的保护范围。
Claims (10)
1.生物基质耦合微生物在制备水凝胶上的应用。
2.权利要求1所述的应用,生物基质包括茶渣、咖啡渣、玉米秸秆、柚子皮、花生壳、草灰、木炭。
3.权利要求1所述的应用,微生物包括酵母菌、细菌、霉菌和微藻类。
4.权利要求3所述的应用,其特征在于:
酵母菌包括库德毕赤酵母、鲁氏酵母、球拟酵母、酿酒酵母、啤酒酵母、圆红冬孢酵母菌、红酵母、红发夫酵母、假丝酵母、汉逊德巴利酵母、接合酵母;
细菌包括大肠杆菌、枯草杆菌、伯克氏菌、蜡样芽孢杆菌、乳酸菌、鞘细菌、荧光假单胞菌;
霉菌包括黑曲霉、米曲霉、红曲霉、酱油曲霉、拟青霉菌、毛霉菌、根霉、黄曲霉、卡伍尔链霉菌。
微藻类:蓝藻、绿藻、金藻、红藻、螺旋藻、雨生红球藻、硅藻。
5.一种水凝胶,其特征在于,包括茶渣和酵母细胞。
6.权利要求5所述的水凝胶的制备方法,其特征在于,包括下列步骤:
(一) 制备酵母菌泥制备;
(二) 茶渣粉水溶液制备;
(三)茶渣水凝胶的制备与处理。
7.权利要求6所述的水凝胶的制备方法,其特征在于,步骤(一)制备酵母菌泥制备的具体步骤为:
将菌种接种到斜面培养基上,在恒温培养箱中培养24h后取出,接种到YPD中,在摇床上扩大培养24h;然后取1mL菌液,分别加入YPD及用NaCl胁迫的YPD中,在摇床上培养24h后,将菌液分别置于5mL的离心管中,3700r离心5min,即得到菌泥,菌泥中菌的数量为3-5×106。
8.权利要求6所述的水凝胶的制备方法,其特征在于,步骤(二)茶渣粉水溶液制备的具体步骤为:
分别配置0.5 mol/L的氢氧化钠溶液和氯化氢溶液,量取0.5 mol/L 的10.5 mL的氢氧化钠溶液,将茶渣粉过100目筛,加入1-5g茶渣粉,混合搅拌1-30 min后加入0.5 mol/L 的10.5 mL盐酸溶液调至中性,得到茶渣水溶液。
9.权利要求6所述的水凝胶的制备方法,其特征在于,步骤(三)茶渣水凝胶的制备与处理的具体步骤为:
茶渣水溶液中加入0.3 g过硫酸铵,0.1215 g N,N-亚甲基双丙烯酰胺,3.6 mL丙烯酸和5 mL菌液,菌液为菌泥加去离子水重新稀释至5mL变成菌液;
搅拌混匀后,分装到模具中,每个模具中含有5 mL混合物,充入氮气密封,在60℃条件下水浴3-8 h;
取出反应结束的水凝胶,于去离子水中充分透析24h,每隔2-6 h换一次水;透析结束后,取出水凝胶冻干,备用。
10.权利要求1-5任一项所述的水凝胶在吸附重金属上的应用。
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