CN115232766A - 一种嗜盐微生物污水清洗剂及其制备方法 - Google Patents
一种嗜盐微生物污水清洗剂及其制备方法 Download PDFInfo
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Abstract
本发明涉及一种嗜盐微生物污水清洗剂及其制备方法,涉及微生物发酵及污水处理领域。该方法包括:将从韩国黄海中分离得到嗜盐微生物进行菌株液体培养,在温度条件为30℃,pH7.0、3%盐浓度下培养24小时。取菌培上清液,加入混合有机溶剂进行沉淀。对沉淀物冷冻真空干燥,将干燥后的固体进行粉碎研磨,获粒径为200nm‑100μm的嗜盐微生物清洗剂。本发明制备的嗜盐微生物清洗剂具有高效地生物降解能力,具有良好的经济和环保效益,对污水中存在的动物排泄物进行无害化处理,并且处理过程中不会产生二次污染。
Description
技术领域
本发明涉及一种嗜盐微生物污水清洗剂及其制备方法,涉及微生物发酵及污水处理领域。
背景技术
众所周知,家禽、畜牧业等中大型养殖场每天排放大量的废水,并且废水中含有较多的污染物,例如畜禽粪便,畜禽药物残留,畜禽病原体等。如果没有经过污水处理,直接排放畜牧养殖废水将造成严重的生态环境污染,同时废水被再次利用与农田或生活领域将形成更为严峻的农田污染或危害。同时,一些大型纺织厂,污水处理厂等工厂排放废水时伴随严重的污水异味,严重影响周围环境和生活活动。污染废水中伴随产生的异味主要是由一些硫化合物或者氮化合物等物质产生,且是强烈刺激性的异味,可通过不同途径进入人体,如呼吸道、眼睛、皮肤等,使人头晕不舒服,长时间放置,对人体神经系统会产生严重的影响。
目前已经发现的自然生物处理法是一种利用天然的水体和土壤中的微生物来净化废水的方法,亦称,主要有水体净化和土壤净化两种。在有氧化塘、养殖塘和土地处理等领域受到了广泛的应用。虽然自然生物处理法费用低,对一些难生化降解的有机物、氮磷等营养物和细菌的有较为明显的去除功效。但建立自然生物处理基地占地面积大且其处理效率易受季节等因素的影响。
微生物污水清洗剂是通过微生物菌株发酵过程中产生具有除污活性功能的代谢产物组成,其主要功能的生物大分子具有良好的生物降解能力,并且无毒,可有效去除生活污水中的悬浮物BOD和COD。目前,未见有关本发明所用嗜盐微生物菌种(Sphingorhabdussp.M375)生产微生物清洗剂的相关报道,且在专利检索中未发现微生物清洗剂在高盐浓度(1-12%NaCl)情况下使用的专利。
发明内容
本发明的目的旨在提供一种利用嗜盐微生物菌种发酵后的菌液进行污水处理清洗剂功效的制备方法,该方法具有良好的生物降解能力,成本低廉的且绿色无污染,有效去除污水中的悬浮物,BOD和COD,尤其对畜禽粪便具有明显功效。
黄海鞘氨醇杆状菌为(Sphingorhabdus sp.M375):革兰氏阳性菌,菌落成凸起且有光泽的乳白色状,可通过鞭毛带动运动。该菌株可以在NaCl浓度为1-17%(w/v)(最佳,3%),温度为10-40℃(最佳,30℃)和pH6.0-11.0(最佳,pH7.0)的环境条件下生长。具有氧化酶和过氧化氢酶阳性活性。具有七叶皂苷水解,硝酸盐还原为亚硝酸盐的能力。由d-葡萄糖,l-阿拉伯糖(弱),d-甘露糖(弱),d-甘露醇,N-乙酰葡糖胺(弱),麦芽糖,葡萄糖酸钾,己二酸,苹果酸和柠檬酸三钠产生酸。酯酶(C4),亮氨酸芳基酰胺酶,缬氨酸芳基酰胺酶,酸性磷酸酶(弱),碱性磷酸酶(弱),萘酚作为双水解酶和α-葡萄糖苷酶的活性均为阳性。该型菌株在基因组DNA中具有55.4mol%G+C的含量。
本发明通过以下技术方案实现上述目的:
本发明中涉及的微生物菌株已于2021年3月8日保藏于韩国国家农业生物技术研究所农业微生物菌种保藏,保藏中心登记入册编号为KACC22163。
为实现本发明的目的,将嗜盐微生物按1:100的比例添加到培养液中,置于摇床上进行培养,从而获得黄海鞘氨醇杆状菌培养液;黄海鞘氨醇杆状菌培养基成分为氯化钠19.45g/L,硼酸三月桂酯0.02g,氯化镁8.8g/L,蛋白胨5.0g/L,Na2SO3 3.24g/L,CaCl21.8g/L,硅酸钠4.0mg/L,酵母提取物1.0g,氯化钾0.55g/L,硝酸铵1.6mg/L,碳酸氢钠0.16g/L,磷酸氢二钠8.0mg/L,柠檬酸铁0.1g/L,溴化钾0.08g/L,二氯化锶0.03g/L,氟化钠2.4mg/L,pH7.0±0.3。菌株培养温度优选为30℃。所述培养时间优选为24h。摇床转速为160r/min。
所述的黄海鞘氨醇杆状菌培养液为黄海鞘氨醇杆状菌(Sphingorhabdussp.M375)在黄海鞘氨醇杆状菌培养基内培养24h后,进行8000g离心15分钟所得到的菌培上清液。
黄海鞘氨醇杆状菌清洗剂的制备:向菌培上清中加入体积比为10:1的比例混合无水乙醇和异丙醇制备而成的有机溶剂混合液,按菌培上清:混合有机溶剂为1:2的比例进行混合进行沉淀,4℃放置过夜,再经过10000g,10min离心后弃上清,将沉淀物置于冻干机内冻干,将冻干后的固体进行粉碎研磨,制成为200nm-100μm的纯品嗜盐微生物清洗剂。
所述的制备方法包含以下步骤:
S1、按照权利要求3所述嗜盐微生物清洗剂培养液制备成分及其重量称取各成分制成培养液后接种黄海鞘氨醇杆状菌(Sphingorhabdus sp.M375)在30℃条件下培养24-36小时;
S2、按照权利要求4所述嗜盐微生物清洗剂的制备成分及其重量份数取各原料;
S3、向步骤1所得到的菌培上清加入步骤2中制备好的有机溶剂混合液后,在4℃放置过夜,离心后得到的沉淀进行冻干处理。
S4、将步骤3中获得的成品粉末进行500倍稀释后制得嗜盐微生物清洗剂成品。
本发明的有益效果是:本制备方法在制备过程中所使用的材料及产生的物质均不会造成二次污染,使用安全,易于推广。
本专利制备的嗜盐微生物清洗剂应用场景广泛,适用领域也广泛覆盖从低盐浓度到高盐浓度污水,易于广泛应用与各种环境条件。
附图说明
图1为本发明实施例不同盐浓度条件对BOD清除率的影响,在图1中,横坐标为盐浓度(w/v),纵坐标为BOD清除率(%);
图2为本发明实施例不同温度范围对BOD清除率的影响,在图2中,横坐标为温度(℃)范围,纵坐标为BOD清除率(%)。
具体实施方式
下面结合实施例对本发明做进一步详细描述。
实施例1
嗜盐微生物清洗剂的制备
将嗜盐微生物按1:100的比例添加到到装有500mL培养基的1L三角瓶中培养液中,置于于30℃、160r/min的摇床上进行24小时的培养,从而获得黄海鞘氨醇杆状菌培养液。
黄海鞘氨醇杆状菌培养基成分氯化钠19.45g/L,硼酸三月桂酯0.02g,氯化镁8.8g/L,蛋白胨5.0g/L,Na2SO3 3.24g/L,CaCl2 1.8g/L,硅酸钠4.0mg/L,酵母提取物1.0g,氯化钾0.55g/L,硝酸铵1.6mg/L,碳酸氢钠0.16g/L,磷酸氢二钠8.0mg/L,柠檬酸铁0.1g/L,溴化钾0.08g/L,二氯化锶0.03g/L,氟化钠2.4mg/L,pH7.0±0.3。本实施方式中涉及的所有培养基均在121℃下灭菌15分钟。冷却到室温后,接种黄海鞘氨醇杆状菌。培养24h后,进行8000g离心15分钟所得到的菌培上清液。
向菌培上清中加入预制的体积比为10:1混合的无水乙醇和异丙醇有机溶剂混合液,按菌培上清:混合有机溶剂为1:2的比例进行混合进行沉淀,4℃放置过夜,再经过10000g,10min离心后弃上清,将沉淀物置于冻干机内冻干,将冻干后的固体进行粉碎研磨,制成为200nm-100μm的纯品嗜盐微生物清洗剂。
嗜盐微生物清洗剂清洗畜禽废水
将含有黄海鞘氨醇杆状菌清洗剂10mL,加入沉淀物100mL,在30℃的室温环境下,置于摇床上震荡,在实验开始的第3天分别测定样品的BOD,计算得该清洗剂的BOD清除率为97.3%,降解率为100%。
嗜盐微生物清洗剂在不同盐浓度范围内清洗畜禽废水
将含有黄海鞘氨醇杆状菌清洗剂10mL,加入沉淀物100mL,在30℃,盐浓度0-21%(w/v)环境下,置于摇床上180rpm震荡培养,在实验开始的第3天分别测定样品的BOD,计算得该清洗剂的BOD清除率为97.3%,降解率为100%。图1表明黄海鞘氨醇杆状菌清洗剂对盐浓度3-5%(w/v)范围内的污水有较好的处理效果。
嗜盐微生物清洗剂在不同温度条件下清洗畜禽废水
将含有黄海鞘氨醇杆状菌清洗剂10mL,加入沉淀物100mL,在4-40℃的摇床条件下180rpm震荡培养,在实验开始的第3天分别测定样品的BOD,计算得该清洗剂的BOD清除率为97.3%,降解率为100%。图2表明黄海鞘氨醇杆状菌清洗剂对温度25-30范围内的污水有较好的处理效果。
以上所述仅是本发明的优选实施方式,应当指出,对于本技术领域的普通技术人员来说,在不脱离本发明原理的前提下,还可以做出若干改进和润饰,这些改进和润饰也应视为本发明的保护范围。
Claims (9)
1.一种清洁污水的黄海鞘氨醇杆状菌(Sphingorhabdus sp.)菌株M375,其特征在于,保藏中心登记入册编号为KACC 22163。
2.权利要求1所述M375的培养方法,其特征在于,包括以下步骤:将所述M375于培养液中振荡培养24~36h;
所述培养液包括以下浓度的成分:氯化钠19.45g/L、硼酸三月桂酯0.02g、氯化镁8.8g/L、蛋白胨5.0g/L、Na2SO3 3.24g/L、CaCl2 1.8g/L、硅酸钠4.0mg/L、酵母提取物1.0g、氯化钾0.55g/L、硝酸铵1.6mg/L、碳酸氢钠0.16g/L、磷酸氢二钠8.0mg/L、柠檬酸铁0.1g/L、溴化钾0.08g/L、二氯化锶0.03g/L和氟化钠2.4mg/L,pH7.0±0.3。
3.根据权利要求2所述培养方法,其特征在于,所述振荡培养的温度为30℃,摇床转速为160rpm。
4.一种黄海鞘氨醇杆状菌清洗剂,其特征在于,包括权利要求1所述M375的培养上清液和有机溶剂。
5.根据权利要求4所述清洗剂,其特征在于,所述有机溶剂包括无水乙醇和异丙醇;所述无水乙醇和异丙醇的体积比为10:1。
6.根据权利要求4所述清洗剂,其特征在于,所述培养上清液和有机溶剂的体积比为1:2。
7.权利要求4~6任一项所述清洗剂的制备方法,其特征在于,包括以下步骤:将权利要求2或3所述培养方法得到的菌培液,8000g离心15分钟,收集上清液;
将所述上清液与有机溶剂进行混合,4℃放置过夜,再经过10000g 10min离心后弃上清。
8.根据权利要求7所述制备方法,其特征在于,在弃上清后,还包括对沉淀物置于冻干机内冻干,将冻干后的固体进行粉碎研磨,制成为粒径200nm-100μm的纯品嗜盐微生物清洗剂。
9.权利要求4~6任一项所述清洗剂在去除污水废水中的畜禽粪便并且有效消除污水中的异味中的应用。
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