CN114196654A - 一种复合酶制剂及其制备方法和产生沼气的方法 - Google Patents
一种复合酶制剂及其制备方法和产生沼气的方法 Download PDFInfo
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- CN114196654A CN114196654A CN202111553877.XA CN202111553877A CN114196654A CN 114196654 A CN114196654 A CN 114196654A CN 202111553877 A CN202111553877 A CN 202111553877A CN 114196654 A CN114196654 A CN 114196654A
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- enzyme preparation
- complex enzyme
- treated
- amylase
- parts
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Abstract
本发明公开了一种复合酶制剂:淀粉酶20‑30份、纤维素酶20‑30份、木聚糖酶15‑25份、脂肪酶5‑10份和果胶酶5‑10份,其中,淀粉酶为α‑淀粉酶、β‑淀粉酶、葡萄糖淀粉酶和淀粉‑1,6‑葡萄糖苷酶中的至少一种;复合酶制剂的制备方法:称取各原料后搅拌;产生沼气的方法:将禽畜粪便、农作物秸秆、餐厨垃圾和蔬菜废弃物处理后搅拌混合均匀,加入水、复合酶制剂和硝酸亚铁,厌氧发酵,即得。本发明从沼气厌氧发酵微生物代谢及酶活变化规律的角度出发,通过复合酶制剂配方的使用,达到促进发酵原料快速、高效水解及沼气发酵微生物生长代谢的效果,进而实现发酵原料厌氧发酵产沼气系统高效、稳定运行的目的。
Description
技术领域
本发明涉及可再生能源利用及环境保护技术领域,更具体的说是涉及一种复合酶制剂及其制备方法和产生沼气的方法。
背景技术
随着煤炭、石油、天然气的长期使用,这些不可再生的化石能源正在面临着枯竭的危险。与此同时,我国的农业生产中每年都有大量作物秸秆、根、茎等废弃物,畜牧生产大量动物粪便,以及人类生产生活中的大量有机废物。
沼气技术作为一个以禽畜粪便、作物秸秆、食品加工废物和废水以及酒精废料等有机垃圾为原料生产甲烷等可燃性气体的技术正得到越来越多人的重视。利用有机物厌氧发酵生产沼气是解决能源短缺,同时减少有机废弃物的有效方法。
对于村镇量大面广的生物质有机废弃物而言,常见的村镇有机固体废弃物主要包括农作物秸秆、餐厨垃圾、禽畜粪便和蔬菜废弃物。由于受发酵原料本身营养特性及结构等方面的限制,单一原料发酵产沼气的局限性越来越突出。如以单一秸秆为发酵原料,由于秸秆碳氮比高、纤维素含量高、营养元素缺乏,导致原料厌氧发酵消化周期长、降解率低;粪便原料中氨氮含量高,单一的粪便厌氧发酵,氨氮会对甲烷菌和产氢菌有一定的抑制作用;单一以餐厨、蔬菜为发酵原料,这些原料中含有多种易降解成分,发酵速度快,易酸化,致使产气性能降低,甚至导致厌氧消化失败。同时,由于沼气技术受到原料结构以及菌种生长环境等方面的制约,产气速度慢,产气少,产气时间短,原料利用率低,有大量的残留能量等问题。
近年来,生物技术尤其是生物酶技术的快速发展,为我们在解决沼气生产中的诸多问题提供了一个突破口。在正常沼气发酵过程中,发酵原料的厌氧分解经历了水解、专性产氢产酸和产甲烷三个阶段,其中水解过程是沼气发酵的开始,也是沼气发酵的限速步骤。相关的研究报道表明,酶制剂的酶活在沼气发酵过程中与沼气的产气量有密切的关系。若能在沼气发酵体系中保持一定的酶活水平,可能有利于提高沼气的产量。
但是,目前市面上可用于大规模产生沼气的酶制剂却少之又少,而且大多存在在发酵第一阶段效率低、反应不充分的问题,效果不是特别理想。
因此,如何开发一种用于产生沼气的酶制剂和方法是本领域技术人员亟需解决的问题。
发明内容
有鉴于此,本发明的目的在于提供一种复合酶制剂及其制备方法和产生沼气的方法,以解决现有技术中的不足。
为了实现上述目的,本发明采用如下技术方案:
复合酶制剂,包括以下重量份的原料:淀粉酶20-30份、纤维素酶20-30份、木聚糖酶15-25份、脂肪酶5-10份和果胶酶5-10份;其中,淀粉酶为α-淀粉酶、β-淀粉酶、葡萄糖淀粉酶和淀粉-1,6-葡萄糖苷酶中的至少一种。
本发明的有益效果在于,本发明所选淀粉酶具有较高的酶活性,从而显著提高沼气发酵速率和产气量。
进一步,上述纤维素酶为外切β-葡聚糖酶、内切β-葡聚糖酶和β-葡萄糖苷酶中的至少一种。
采用上述进一步技术方案的有益效果在于,本发明所选纤维素酶能够作用于纤维素以及从纤维素衍生出来的产物,且产量高,活性大。
进一步,上述木聚糖酶为β-1,4-内切木聚糖酶、β-木糖苷酶、α-L-阿拉伯糖苷酶、α-D-葡糖苷酸酶、乙酰基木聚糖酶和酚酸酯酶中的至少一种。
采用上述进一步技术方案的有益效果在于,本发明所选木聚糖酶通过其中各种组分的相互协同作用来降解木聚糖,它们作用于木糖与侧链取代基之间的糖苷键,协同主链水解酶的作用,最终将木聚糖转化为它的组成单糖。
进一步,上述脂肪酶为磷酸酯酶、固醇酶和羧酸酯酶中的至少一种。
采用上述进一步技术方案的有益效果在于,本发明所选脂肪酶具有油-水界面的亲和力,能在油-水界面上高速率的催化水解不溶于水的脂类物质,且具有更广的作用pH、作用温度范围、高稳定性和高活性,对底物有特异性。
进一步,上述果胶酶为聚甲基半乳糖、醛酸酶、聚半乳糖醛酸酶和聚半乳糖醛酸裂解酶中的至少一种。
采用上述进一步技术方案的有益效果在于,本发明所选果胶酶能够使细胞间的果胶质降解,把细胞从组织内分离出来。
上述复合酶制剂的制备方法,具体包括以下步骤:
(1)按上述复合酶制剂的重量份数称取各原料;
(2)将淀粉酶、纤维素酶、木聚糖酶、脂肪酶和果胶酶混合后搅拌,即得复合酶制剂。
进一步,上述步骤(2)中,搅拌的速度为100-200r/min,时间为40-60min。
上述复合酶制剂产生沼气的方法,具体包括以下步骤:
(1)收集禽畜粪便、农作物秸秆、餐厨垃圾和蔬菜废弃物;
(2)将农作物秸秆粉碎,然后加入到石灰水中搅拌均匀并浸泡,得到处理后的农作物秸秆,备用;
(3)将餐厨垃圾去浮油后与蔬菜废弃物混合,然后搅碎,得到处理后的餐厨垃圾和处理后的蔬菜废弃物,备用;
(4)将禽畜粪便、处理后的农作物秸秆、处理后的餐厨垃圾和处理后的蔬菜废弃物搅拌混合均匀,得到混合料;
(5)向混合料中加入水、复合酶制剂和硝酸亚铁搅拌混合均匀,得到发酵原料;
(6)将发酵原料加入发酵池内进行厌氧发酵,即得沼气。
进一步,上述步骤(1)中,禽畜粪便为猪粪、牛粪、羊粪和鸡粪中的至少一种,农作物秸秆为小麦秸秆、玉米秸秆、水稻秸秆和高粱秸秆中的至少一种,蔬菜废弃物为为马铃薯秧、黄瓜藤蔓、辣椒秧、西红柿秧、白菜叶和甘蓝菜叶中的至少一种。
进一步,上述步骤(2)中,粉碎至粒径<1cm,石灰水的质量为农作物秸秆质量的2-3倍,浸泡的时间为1-2天;
进一步,上述步骤(3)中,搅碎至粒径<1cm;
进一步,上述步骤(4)中,禽畜粪便的含水率为50%-60%,处理后的农作物秸秆的含水率为20%-30%,处理后的餐厨垃圾的含水率为55%-65%,处理后的蔬菜废弃物的含水率为30%-40%,禽畜粪便、处理后的农作物秸秆、处理后的餐厨垃圾和处理后的蔬菜废弃物的质量比为(20-40):(10-20):(10-20):(10-20);
进一步,上述步骤(5)中,水的加入量为混合料质量的1-2倍,复合酶制剂的加入量为混合料质量的0.2%-0.8%,硝酸亚铁的加入量为混合料质量的0.5%-1.5%。
采用上述进一步技术方案的有益效果在于,硝酸亚铁能够为整个发酵过程提供微量元素,为产甲烷菌提供营养,促进甲烷菌的生长加快,改善发酵池内生物菌种的活性变化,进而提高有机物的降解率。
进一步,上述步骤(6)中,厌氧发酵温度为38-45℃,时间为30-35天。
经由上述的技术方案可知,与现有技术相比,本发明的有益效果如下:
1、通过配比之后的复合酶制剂各组分间存在协同作用,能够有效降解原料中的有机物,从而解决大分子有机物在发酵第一阶段效率低、反应不充分的问题,进而使沼气产气量得到明显的提高,发酵速率加快。
2、本发明复合酶制剂能够将发酵原料中的各种大分子有机物全部或部分降解为小分子,破坏原料的组织结构,利于菌种的利用,使快速增殖,从而达到快速产气、延长细菌生长期、提高沼气产量和提高原料利用率的效果。
3、本发明从沼气厌氧发酵微生物代谢及酶活变化规律的角度出发,通过复合酶制剂配方的使用,达到促进发酵原料快速、高效水解及沼气发酵微生物生长代谢的效果,进而实现发酵原料厌氧发酵产沼气系统高效、稳定运行的目的。
具体实施方式
下面对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例仅仅是本发明一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例,都属于本发明保护的范围。
实施例1
复合酶制剂,包括以下原料:淀粉酶25kg、纤维素酶25kg、木聚糖酶20kg、脂肪酶8kg和果胶酶8kg;
其中,淀粉酶为α-淀粉酶、β-淀粉酶、葡萄糖淀粉酶和淀粉-1,6-葡萄糖苷酶以等质量混合后的的混合物,纤维素酶为外切β-葡聚糖酶、内切β-葡聚糖酶和β-葡萄糖苷酶以等质量混合后的的混合物,木聚糖酶为β-1,4-内切木聚糖酶、β-木糖苷酶、α-L-阿拉伯糖苷酶、α-D-葡糖苷酸酶、乙酰基木聚糖酶和酚酸酯酶以等质量混合后的的混合物,脂肪酶为磷酸酯酶、固醇酶和羧酸酯酶以等质量混合后的的混合物,果胶酶为聚甲基半乳糖、醛酸酶、聚半乳糖醛酸酶和聚半乳糖醛酸裂解酶以等质量混合后的的混合物;
复合酶制剂的制备方法,具体包括以下步骤:
(1)按上述复合酶制剂的重量称取各原料;
(2)将淀粉酶、纤维素酶、木聚糖酶、脂肪酶和果胶酶加入反应釜中,混合后以150r/min的速度搅拌50min,即得复合酶制剂;
复合酶制剂产生沼气的方法,具体包括以下步骤:
(1)收集禽畜粪便、农作物秸秆、餐厨垃圾和蔬菜废弃物;
其中,禽畜粪便为猪粪、牛粪、羊粪和鸡粪以等质量混合后的的混合物,农作物秸秆为小麦秸秆、玉米秸秆、水稻秸秆和高粱秸秆以等质量混合后的的混合物,蔬菜废弃物为为马铃薯秧、黄瓜藤蔓、辣椒秧、西红柿秧、白菜叶和甘蓝菜叶以等质量混合后的的混合物;
(2)将农作物秸秆粉碎至粒径<1cm,然后加入到3倍质量的石灰水中搅拌均匀并浸泡1天,得到处理后的农作物秸秆,备用;
(3)将餐厨垃圾去浮油后与蔬菜废弃物混合,然后搅碎至粒径<1cm,得到处理后的餐厨垃圾和处理后的蔬菜废弃物,备用;
(4)将禽畜粪便(含水率为55%)40kg、处理后的农作物秸秆(含水率为25%)20kg、处理后的餐厨垃圾(含水率为60%)20kg和处理后的蔬菜废弃物(含水率为35%)20kg搅拌混合均匀,得到混合料;
(5)向混合料中加入水412.5kg、复合酶制剂0.5kg和硝酸亚铁1kg搅拌混合均匀,得到干物质浓度(TS)为10%的发酵原料460kg;
(6)将干物质浓度(TS)为10%的发酵原料460kg加入池容为600L的发酵池内,调节pH值为7,在42℃温度下厌氧发酵30天,即得沼气。
实施例2
复合酶制剂,包括以下原料:淀粉酶20kg、纤维素酶30kg、木聚糖酶15kg、脂肪酶10kg和果胶酶10kg;
其中,淀粉酶为α-淀粉酶、β-淀粉酶、葡萄糖淀粉酶和淀粉-1,6-葡萄糖苷酶以等质量混合后的的混合物,纤维素酶为外切β-葡聚糖酶、内切β-葡聚糖酶和β-葡萄糖苷酶以等质量混合后的的混合物,木聚糖酶为β-1,4-内切木聚糖酶、β-木糖苷酶、α-L-阿拉伯糖苷酶、α-D-葡糖苷酸酶、乙酰基木聚糖酶和酚酸酯酶以等质量混合后的的混合物,脂肪酶为磷酸酯酶、固醇酶和羧酸酯酶以等质量混合后的的混合物,果胶酶为聚甲基半乳糖、醛酸酶、聚半乳糖醛酸酶和聚半乳糖醛酸裂解酶以等质量混合后的的混合物;
复合酶制剂的制备方法,具体包括以下步骤:
(1)按上述复合酶制剂的重量称取各原料;
(2)将淀粉酶、纤维素酶、木聚糖酶、脂肪酶和果胶酶加入反应釜中,混合后以100r/min的速度搅拌60min,即得复合酶制剂;
复合酶制剂产生沼气的方法,具体包括以下步骤:
(1)收集禽畜粪便、农作物秸秆、餐厨垃圾和蔬菜废弃物;
其中,禽畜粪便为猪粪、牛粪、羊粪和鸡粪以等质量混合后的的混合物,农作物秸秆为小麦秸秆、玉米秸秆、水稻秸秆和高粱秸秆以等质量混合后的的混合物,蔬菜废弃物为为马铃薯秧、黄瓜藤蔓、辣椒秧、西红柿秧、白菜叶和甘蓝菜叶以等质量混合后的的混合物;
(2)将农作物秸秆粉碎至粒径<1cm,然后加入到2倍质量的石灰水中搅拌均匀并浸泡2天,得到处理后的农作物秸秆,备用;
(3)将餐厨垃圾去浮油后与蔬菜废弃物混合,然后搅碎至粒径<1cm,得到处理后的餐厨垃圾和处理后的蔬菜废弃物,备用;
(4)将禽畜粪便40kg(含水率为50%)、处理后的农作物秸秆(含水率为20%)20kg、处理后的餐厨垃圾(含水率为55%)20kg和处理后的蔬菜废弃物(含水率为30%)20kg搅拌混合均匀,得到混合料;
(5)向混合料中加入水470.8kg、复合酶制剂0.2kg和硝酸亚铁0.5kg搅拌混合均匀,得到干物质浓度(TS)为8%的发酵原料512.5kg;
(6)将干物质浓度(TS)为8%的发酵原料512.5kg加入池容为600L的发酵池内,调节pH值为7,在38℃温度下厌氧发酵30天,即得沼气。
实施例3
复合酶制剂,包括以下原料:淀粉酶30kg、纤维素酶20kg、木聚糖酶15kg、脂肪酶10kg和果胶酶10kg;
其中,淀粉酶为α-淀粉酶、β-淀粉酶、葡萄糖淀粉酶和淀粉-1,6-葡萄糖苷酶以等质量混合后的的混合物,纤维素酶为外切β-葡聚糖酶、内切β-葡聚糖酶和β-葡萄糖苷酶以等质量混合后的的混合物,木聚糖酶为β-1,4-内切木聚糖酶、β-木糖苷酶、α-L-阿拉伯糖苷酶、α-D-葡糖苷酸酶、乙酰基木聚糖酶和酚酸酯酶以等质量混合后的的混合物,脂肪酶为磷酸酯酶、固醇酶和羧酸酯酶以等质量混合后的的混合物,果胶酶为聚甲基半乳糖、醛酸酶、聚半乳糖醛酸酶和聚半乳糖醛酸裂解酶以等质量混合后的的混合物;
复合酶制剂的制备方法,具体包括以下步骤:
(1)按上述复合酶制剂的重量称取各原料;
(2)将淀粉酶、纤维素酶、木聚糖酶、脂肪酶和果胶酶加入反应釜中,混合后以200r/min的速度搅拌40min,即得复合酶制剂;
复合酶制剂产生沼气的方法,具体包括以下步骤:
(1)收集禽畜粪便、农作物秸秆、餐厨垃圾和蔬菜废弃物;
其中,禽畜粪便为猪粪、牛粪、羊粪和鸡粪以等质量混合后的的混合物,农作物秸秆为小麦秸秆、玉米秸秆、水稻秸秆和高粱秸秆以等质量混合后的的混合物,蔬菜废弃物为为马铃薯秧、黄瓜藤蔓、辣椒秧、西红柿秧、白菜叶和甘蓝菜叶以等质量混合后的的混合物;
(2)将农作物秸秆粉碎至粒径<1cm,然后加入到3倍质量的石灰水中搅拌均匀并浸泡2天,得到处理后的农作物秸秆,备用;
(3)将餐厨垃圾去浮油后与蔬菜废弃物混合,然后搅碎至粒径<1cm,得到处理后的餐厨垃圾和处理后的蔬菜废弃物,备用;
(4)将禽畜粪便(含水率为60%)40kg、处理后的农作物秸秆(含水率为30%)20kg、处理后的餐厨垃圾(含水率为65%)20kg和处理后的蔬菜废弃物(含水率为40%)20kg搅拌混合均匀,得到混合料;
(5)向混合料中加入水456.7kg、复合酶制剂0.8kg和硝酸亚铁1.5kg搅拌混合均匀,得到干物质浓度(TS)为10%的发酵原料510kg;
(6)将干物质浓度(TS)为10%的发酵原料510kg加入池容为600L的发酵池内,调节pH值为7,在45℃温度下厌氧发酵30天,即得沼气。
对比例
与实施例1的区别仅在于,产生沼气的步骤中不添加复合酶制剂。
性能测试
分别按照实施例1-3中复合酶制剂产生沼气的方法生产沼气,同时以对比例作为对照,以每吨混合料干物质量TS计,分别计算其30天总产气量(m3/t)。
结果如表1所示。
表1实施例1-3和对比例沼气产率
沼气 | 30天总产气量(m<sup>3</sup>/t) |
实施例1 | 386 |
实施例2 | 382 |
实施例3 | 379 |
对比例 | 212 |
由表1可知,与对比例相比,本发明实施例1-3的30天总产气量均显著提高。
以上试验说明,本发明通过配比之后的复合酶制剂各组分间存在协同作用,能够有效降解原料中的有机物,从而解决大分子有机物在发酵第一阶段效率低、反应不充分的问题,进而使沼气产气量得到明显的提高,发酵速率加快。
对所公开的实施例的上述说明,使本领域专业技术人员能够实现或使用本发明。对这些实施例的多种修改对本领域的专业技术人员来说将是显而易见的,本文中所定义的一般原理可以在不脱离本发明的精神或范围的情况下,在其它实施例中实现。因此,本发明将不会被限制于本文所示的这些实施例,而是要符合与本文所公开的原理和新颖特点相一致的最宽的范围。
Claims (10)
1.一种复合酶制剂,其特征在于,包括以下重量份的原料:淀粉酶20-30份、纤维素酶20-30份、木聚糖酶15-25份、脂肪酶5-10份和果胶酶5-10份;
所述淀粉酶为α-淀粉酶、β-淀粉酶、葡萄糖淀粉酶和淀粉-1,6-葡萄糖苷酶中的至少一种。
2.根据权利要求1所述的一种复合酶制剂,其特征在于,所述纤维素酶为外切β-葡聚糖酶、内切β-葡聚糖酶和β-葡萄糖苷酶中的至少一种。
3.根据权利要求1所述的一种复合酶制剂,其特征在于,所述木聚糖酶为β-1,4-内切木聚糖酶、β-木糖苷酶、α-L-阿拉伯糖苷酶、α-D-葡糖苷酸酶、乙酰基木聚糖酶和酚酸酯酶中的至少一种。
4.根据权利要求1所述的一种复合酶制剂,其特征在于,所述脂肪酶为磷酸酯酶、固醇酶和羧酸酯酶中的至少一种。
5.根据权利要求1所述的一种复合酶制剂,其特征在于,所述果胶酶为聚甲基半乳糖、醛酸酶、聚半乳糖醛酸酶和聚半乳糖醛酸裂解酶中的至少一种。
6.一种复合酶制剂的制备方法,其特征在于,具体包括以下步骤:
(1)按权利要求1-5任一项所述复合酶制剂的重量份数称取各原料;
(2)将淀粉酶、纤维素酶、木聚糖酶、脂肪酶和果胶酶混合后搅拌,即得所述复合酶制剂。
7.根据权利要求6所述的一种复合酶制剂的制备方法,其特征在于,步骤(2)中,所述搅拌的速度为100-200r/min,时间为40-60min。
8.一种如权利要求1-5任一项所述复合酶制剂产生沼气的方法,其特征在于,具体包括以下步骤:
(1)收集禽畜粪便、农作物秸秆、餐厨垃圾和蔬菜废弃物;
(2)将农作物秸秆粉碎,然后加入到石灰水中搅拌均匀并浸泡,得到处理后的农作物秸秆,备用;
(3)将餐厨垃圾去浮油后与蔬菜废弃物混合,然后搅碎,得到处理后的餐厨垃圾和处理后的蔬菜废弃物,备用;
(4)将禽畜粪便、处理后的农作物秸秆、处理后的餐厨垃圾和处理后的蔬菜废弃物搅拌混合均匀,得到混合料;
(5)向混合料中加入水、权利要求1-5任一项所述复合酶制剂和硝酸亚铁搅拌混合均匀,得到发酵原料;
(6)将发酵原料加入发酵池内进行厌氧发酵,即得所述沼气。
9.根据权利要求8所述的一种复合酶制剂产生沼气的方法,其特征在于,步骤(1)中,所述禽畜粪便为猪粪、牛粪、羊粪和鸡粪中的至少一种,所述农作物秸秆为小麦秸秆、玉米秸秆、水稻秸秆和高粱秸秆中的至少一种,所述蔬菜废弃物为为马铃薯秧、黄瓜藤蔓、辣椒秧、西红柿秧、白菜叶和甘蓝菜叶中的至少一种;
步骤(2)中,所述粉碎至粒径<1cm,所述石灰水的质量为农作物秸秆质量的2-3倍,所述浸泡的时间为1-2天;
步骤(3)中,所述搅碎至粒径<1cm;
步骤(4)中,所述禽畜粪便的含水率为50%-60%,所述处理后的农作物秸秆的含水率为20%-30%,所述处理后的餐厨垃圾的含水率为55%-65%,所述处理后的蔬菜废弃物的含水率为30%-40%,所述禽畜粪便、处理后的农作物秸秆、处理后的餐厨垃圾和处理后的蔬菜废弃物的质量比为(20-40):(10-20):(10-20):(10-20);
步骤(5)中,所述水的加入量为混合料质量的1-2倍,所述复合酶制剂的加入量为混合料质量的0.2%-0.8%,所述硝酸亚铁的加入量为混合料质量的0.5%-1.5%。
10.根据权利要求8所述的一种复合酶制剂产生沼气的方法,其特征在于,步骤(6)中,所述厌氧发酵温度为38-45℃,时间为30-35天。
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