CN111484938A - 一种用褐腐菌处理印染废水时的菌丝固定化方法 - Google Patents
一种用褐腐菌处理印染废水时的菌丝固定化方法 Download PDFInfo
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Abstract
本发明公开了一种印染废水脱色菌株的菌丝固定化方法,涉及木腐菌、固定化和处理工艺,特别涉及一种印染污水脱色木腐真菌的菌丝固定方法。固定化和处理工艺如下:将菌株在100mL液体培养基中静止培养5天,用高速匀浆机破碎后再次扩大静置培养。用高速匀浆机再次破碎后过滤收集菌体,并用自来水洗涤3次去除培养基并收集湿菌体。破碎农业废物(玉米或小麦秸秆)成碎屑后与收集的菌体以重量比1∶1000(菌体湿重∶木屑干重)混合均匀,加入适量磷酸缓冲液充分湿润后用成型机压成1厘米后的薄片,将薄片用3层尼龙网包裹后放入到3厘米高的长方形塑料筐中,将筐放置到28度的生化培养箱(湿度70%)静止培养48小时进行菌株的固定化。将筐固定在好氧生化池的表面,调整位置使污水刚刚淹没塑料筐表面,进行曝气处理。本发明所述固定化方法工艺流程简单,处理条件温和,不需要特殊设备,投资成本低,具有良好的可操作性,不需要化学合成,处理效果稳定,脱色效率显著,环境友好。
Description
技术领域
本发明涉及木腐菌、印染污水脱色处理和工艺流程,特别涉及一种固定化的木腐菌对染料进行降解和脱色的方法及工艺流程。
背景技术
偶氮类染料被大量应用在印染和皮革等工业领域,由于染色的上色工艺流程效率低的缘故,据估计约有20%的染料没有被有效利用从而以污水的形式被释放到环境中。现代工业中使用的大多数偶氮类染料具有致癌、致畸、致突变等嫌疑。由于染料废水具有COD高,稳定性强,色度高,成分复杂,对普通微生物具有毒害作用等特点,给处理染料废水带来相当的难度。目前处理印染废水的方法主要有生物降解法和物理化学处理方法。物理化学方法处理印染污水时通常费用比较昂贵,且不能完全去除降解产物,可能会造成二次污染,效率也不高,因而其应用受到限制。生物处理法尤其是微生物处理法具有操作简单、运行费用低、无二次污染、对环境友好等优点,当然也存在着处理时间长和效率低等问题。微生物处理法主要有以下两种:细菌降解和真菌降解方法。由于细菌多数对偶氮类染料的降解效率都比较低,且存在着降解产物毒性有可能增加的风险,因此微生物降解偶氮染料的理论和应用大都集中在真菌,尤其是丝状真菌领域。而丝状真菌降解染料的许多研究主要集中在利用白腐菌,云芝和其他一些木腐菌领域。应用的主要瓶颈是木腐菌菌生长缓慢,处理周期长,易受细菌污染;白腐真菌产酶条件比较苛刻,需要在氮或碳限制的条件下,而实际染料废水成分复杂,很难满足这些条件。
褐腐菌是木腐菌一种,在降解木质素初期通过形成羟基自由基对难降解物质进行切割降解。所产生的羟基自由基是目前为止氧化性最强的小分子物质,而白腐菌据文献报道是通过分泌各种不同形式的氧化酶完成对难降解产物的分解,而最近文献指出这一结论并不正确,这也是造成白腐菌处理效果不稳定的原因之一。但褐腐菌通过羟基自由基对木素的降解的结论已经非常确实,其在降解农药方面的应用有很多报道,但是对染料的降解还很少。本发明通过农业废物对褐腐菌进行初步固定,经过短时发酵后使菌丝体侵入木质底物的内部,完成固定,而不需要用化学合成的方法进行固定。因此,本发明提供了一种由农业废物固定化褐腐菌并对印染污水进行处理的方法,在无任何前处理的情况下,印染污水的色度降解效果能达到90%左右。
发明内容
本发明的目的在于提供一种印染废水处理菌株的固定化方法和处理工艺,以克服传统污水生物处理工艺中反应器生物量浓度偏低,泥水分离困难,不耐冲击负荷等问题。本发明采取以下技术方案:
(1)种子液制备:将一个长在直径为9cm平板上、菌龄为15天的木腐菌菌丝用2mL无菌水洗涤后转接入100mL PDA液体培养基中,在27℃条件下静置培养6天,用高速匀浆机破碎3次,每次20sec,破碎的菌丝作为种子液;
(2)菌体的扩大培养:将100mL破碎的菌丝体接种到1000mL PDA液体培养基中,27度静置培养6天,用匀浆机破碎3次,所得的菌丝体溶液作为扩大培养产物;
(3)固定化菌丝的制备:破碎的菌丝体溶液用抽滤方法收集,并采用相同的方法用自来水清洗3次,尽量除去培养基成分,再用抽滤方法对菌液进行收集和浓缩;
(4)固定化材料的前处理:将收集的农业废物,比如小麦或者玉米秸秆破碎切割成1厘米长的杆状物;
(5)菌体的固定化:将过滤收集的湿菌体和破碎的农业废物以1∶1000的重量比例进行混合搅拌,加入适量磷酸缓冲液使混合物湿润后,用成型机进行压缩成型,制备成1厘米厚的片状物。片状物用三层尼龙网包裹,放入到中空的塑料筐中。将塑料框连同菌丝和农业废物一起放到27度生化培养箱(70%湿度)培养3天后获得固定到农业废物上的菌丝体。
(6)污水处理:将含有固定化细菌的塑料筐安置在好氧生化池的液体表面,使污水刚好能淹没固定化材料为宜。
有益效果
本发明的优势在于:褐腐菌经农业固定化后,对染料的降解效果稳定,运行管理简单,降解效率明显优于传统方法,并且固定化方法经济环保,不产生二次污染。
具体实施方案
实施例1.
(1)种子液制备:将一个长在直径为9cm平板上、菌龄为15天的木腐菌菌丝用2mL无菌水洗涤后转接入100mL PDA液体培养基中,在27℃条件下静置培养6天,用高速匀浆机破碎3次,每次20sec,破碎的菌丝作为种子液;
(2)菌体的扩大培养:将100mL破碎的菌丝体接种到1000mL PDA液体培养基中,27度静置培养6天,用匀浆机破碎3次,所得的菌丝体溶液作为扩大培养产物;
(3)固定化菌丝的制备:破碎的菌丝体溶液用抽滤方法收集,并采用相同的方法用自来水清洗3次,尽量除去培养基成分,再用抽滤方法对菌液进行收集和浓缩;
(4)固定化材料的前处理:将收集的农业废物,比如小麦或者玉米秸秆破碎切割成1厘米长的杆状物;
(5)菌体的固定化:将过滤收集的湿菌体和破碎的农业废物以1∶1000的重量比例进行混合搅拌,加入适量磷酸缓冲液使混合物湿润后,用成型机进行压缩成型,制备成1厘米厚的片状物。片状物用三层尼龙网包裹,放入到中空的塑料筐中。将塑料框连同菌丝和农业废物一起放到27度生化培养箱(70%湿度)培养3天后获得固定到农业废物上的菌丝体。
(6)污水处理:将含有固定化细菌的塑料筐安置在好氧生化池的液体表面,使污水刚好能淹没固定化材料为宜,曝气处理2-3天。
Claims (5)
1.一种用褐腐菌处理印染废水时的菌丝固定化方法,包括以下步骤:
(1)种子液制备:将一个长在直径为9cm平板上、菌龄为15天的木腐菌菌丝用2mL无菌水洗涤后转接入100mL PDA液体培养基中,在27℃条件下静置培养6天,用高速匀浆机破碎3次,每次20sec,破碎的菌丝作为种子液;
(2)菌体的扩大培养:将100mL破碎的菌丝体接种到1000mL PDA液体培养基中,27度静置培养6天,用匀浆机破碎3次,所得的菌丝体溶液作为扩大培养产物;
(3)固定化菌丝的制备:破碎的菌丝体溶液用抽滤方法收集,并采用相同的方法用自来水清洗3次,尽量除去培养基成分,再用抽滤方法对菌液进行收集和浓缩;
(4)固定化材料的前处理:将收集的农业废物,比如小麦或者玉米秸秆破碎切割成1厘米长的杆状物;
(5)菌体的固定化:将过滤收集的湿菌体和破碎的农业废物以1∶1000的重量比例进行混合搅拌,加入适量磷酸缓冲液使混合物湿润后,用成型机进行压缩成型,制备成1厘米厚的片状物。片状物用三层尼龙网包裹,放入到中空的塑料筐中。将塑料框连同菌丝和农业废物一起放到27度生化培养箱(70%湿度)培养3天后获得固定到农业废物上的菌丝体。
(6)污水处理:将含有固定化细菌的塑料筐安置在好氧生化池的液体表面,使污水刚好能淹没固定化材料为宜。
2.根据权利要求1所述一种印染废水处理菌株固定方法,其特征在于所述步骤(1)的种子液的制备和步骤(2)中菌株扩大培养方法。
3.根据权利要求1所述一种印染废水处理菌株固定方法,其特征在于所述步骤(3)中固定化菌丝的制备方法。
4.根据权利要求1所述一种印染废水处理菌株固定方法,其特征在于所述步骤(4)中的菌丝体的前处理过程和步骤中(5)中的菌丝在农业废物上的固定化过程。
5.根据权利要求1所述一种印染废水处理菌株固定方法,其特征在于所述步骤(6)中的污水处理过程。
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