CN111218490A - 利用氨和助剂对木质纤维素进行预处理的方法 - Google Patents
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- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/14—Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase
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Abstract
本发明公开了一种利用氨和助剂对木质纤维素进行预处理的方法,属于生物炼制领域。所述方法在氨法预处理中,添加少量的助剂,促进木质素脱除和阻止多糖发生脱皮反应,有效提高糖的转化效率,同时氨可以将羧基类化合物转化成更加低毒的酰胺类化合物,残留氨作为微生物发酵所用的氮源,氨气可以回收利用。本发明方法降低木质纤维素转化成目标产品的生产成本,适用于工业化大规模生产。
Description
技术领域
本发明属于生物炼制技术领域,涉及一种利用氨和助剂对木质纤维素进行预处理的方法。
背景技术
木质纤维素作为生物燃料的重要原材料,主要由纤维素、半纤维素和木质素等组成。目前,木质纤维素的预处理方法主要包括氨法预处理、热水预处理、气爆预处理和离子液体预处理。其中氨法预处理是木质纤维素转化成乙醇等产品等的预处理方式之一,能有效的破坏木质纤维素的结构,增加木质纤维素内部的孔隙(Lau M W,Dale B E.Effect ofprimary degradation-reaction products from Ammonia Fiber Expansion(AFEX)-treated corn stover on the growth and fermentation of Escherichia coliKO11..Bioresource Technology,2010,101(20):7849-7855.),但是氨法预处理对木质素作用较小,高底物酶水解时,水解效率还不够理想。
发明内容
本发明的目的在于提供一种利用氨和助剂对木质纤维素进行预处理的方法。该方法将助剂加入氨法预处理过程中,增加了木质素脱除率,提高木质纤维素转化率,并减少了酶的使用量,降低预处理成本。
实现本发明目的的技术方案如下:
利用氨和助剂对木质纤维素进行预处理的方法,包括以下步骤:
将木质纤维素原料和助剂水溶液均匀混合,通入或加入氨,至混合基质呈碱性,氨的质量为木质纤维素干重的0.5~6.0,80℃~140℃下进行氨法预处理,所述的助剂选自蒽醌或蒽醌衍生物、绿氧、甲醇、异丙醇或甲醛。
所述的木质纤维素为采用常规使用的木质纤维素,可以是小麦秸秆、玉米秸秆、农林废料、水稻秸秆、高粱秸秆、大豆秸秆、林业废料、回收木浆纤维、木屑、软木、硬木或动物粪便。
所述的木质纤维素原料采用常规使用的木质纤维素原料,可以是长条状木质纤维素或颗粒状木质纤维素,长条状木质纤维素的长度为0.01mm~10cm,颗粒状木质纤维素的直径为0.5mm~10cm。
优选地,所述的木质纤维素原料在预处理过程中的质量为总质量的10%~60%,以水、助剂、氨和木质纤维素原料的质量总和为总质量。
所述的助剂水溶液中,水为木质纤维素干重的10%~90%,助剂为木质纤维素干重的0.1~2.0%。
所述氨法预处理选自稀氨水预处理,氨纤维***预处理(AFEX),萃取氨预处理(EA),液氨循环预处理(ARP)或液氨预处理(LAT)。
所述的氨选自氨水溶液、液氨或氨气。
优选地,所述的氨法预处理的时间为10min~360min,所述的预处理时间包括升温时间、维持时间及降温时间。
本发明还提供基于上述预处理方法的提高纤维素乙醇转化率的发酵工艺,包括以下步骤:
步骤1,预处理:将木质纤维素原料和助剂水溶液均匀混合,通入或加入氨,至混合基质呈碱性,氨的质量为木质纤维素干重的0.5~6.0,80℃~140℃下进行氨法预处理,所述的助剂选自蒽醌或蒽醌衍生物、绿氧、甲醇、异丙醇或甲醛;
步骤2,水解:在预处理得到的物料中加入水和水解酶,进行水解;
步骤3,发酵:加入发酵菌株,发酵,得到发酵产品。
步骤2中,所述的水解酶选用木质纤维素发酵工艺中常规使用的酶,可以是纤维素酶和/或半纤维素酶,例如果胶酶、木聚糖酶中的一种或者组合。
步骤3中,所述的发酵菌株选用木质纤维素发酵工艺中常规使用的菌株,可以是酵母、细菌以及霉菌等。
步骤3中,按发酵菌株的不同,得到的发酵产品可以是大宗化学品、精细化学品或动物饲料等,例如乙醇、丁醇、丙酮、乙酸、乳酸、脂肪烃、油脂、蛋白、氨基酸等。
与现有技术相比,本发明具有以下优点:
(1)预处理过程中,添加微量的助剂,提高了脱除木质素的效果和选择性,明显提高了木质纤维素糖的转化效率,提高了21%以上。
(2)预处理后残留在原料中的氨可以作为发酵菌株生长的氮源,减少额外添加营养物质的成本。
(3)预处理后的固体无需水洗等操作,简化操作成本和减少糖分的损失,设备利用率高,适应于工业化生产。
附图说明
图1为实施例1中分别采用AAQ和AFEX预处理玉米秸秆24小时酶水解糖浓度结果图。
图2为实施例2中分别采用AAQ和LHW预处理玉米秸秆24小时酶水解糖浓度结果图。
具体实施方式
为了便于理解本发明,下文将结合附图和较佳的实施例例来对本发明做更全面,细致的描述,但本发明的保护范围并不限于以下具体的实施例。
下文所用到的所有专业术语与本领域技术人员通常理解的含义相同,除非另有定义。本文中所使用的专业术语只是为了描述具体事例的目的,并不是旨在限制本发明的保护范围。
除非另有特别说明,本发明中用到的试剂,原料,仪器和设备等均可市场购买得到或者已经有的方法制备得到。
实施例中使用以下的缩写:
AAQ:氨和助剂预处理(Ammonia andanthraquinone),AFEX:氨气***预处理(Ammonia fiber explosion),LHW:热水预处理(Liquid hot water)。
实施例1
分别采用AAQ预处理和AFEX预处理玉米秸酶水解效果的比较,包括以下步骤:
1.原料准备:将农田里的玉米秸秆收集后,自然晾干后粉碎成1-4mm的颗粒。
2.1AAQ预处理:将粉碎后的玉米秸秆置于反应釜中,干物和氨的比例为1:1,含水率60%(基于玉米秸秆的干重),蒽醌的含量为0.6%(基于玉米秸秆的干重),120℃,反应30min。
2.2AFEX预处理:将粉碎后的玉米秸秆置于反应釜中,干物和氨的比例为1:1,含水率60%(基于玉米秸秆的干重),120℃,反应30min。
3.分别将预处理后的秸秆置于酶水解反应器中,底物质量浓度为3%,加入水解酶和柠檬酸-柠檬酸钠缓冲液,在50℃的振荡箱(250rpm)内进行酶解反应,酶水解24小时。
AAQ预处理和AFEX预处理在3%的干物条件下酶水解效果如图1所示。表示3%玉米秸秆酶水解葡萄糖的浓度,表示3%玉米秸秆酶水解木糖的浓度。从图1可以看出玉米秸秆经过AAQ预处理,酶水解24小时后葡萄糖浓度能达到9.3g/L,而玉米秸秆经过AFEX预处理,酶水解24小时后葡萄糖浓度只有7.8g/L。AAQ预处理玉米秸秆酶水解相比于AFEX,葡萄糖的转化率额提高了29.5%。AAQ预处理处理玉米秸秆酶水解24小时木糖浓度3.4g/L,AFEX预处理处理秸秆木糖浓度为3.0g/L。
实施例2
AAQ预处理和LHW预处理玉米秸酶水解效果的比较,包括以下步骤:
1.原料准备:将农田里的玉米秸秆收集后,自然晾干后粉碎成1-4mm的颗粒。
2.1 AAQ预处理:将粉碎后的玉米秸秆置于反应釜中,干物和氨的比例为1:1,含水率60%(基于玉米秸秆的干重),蒽醌的含量为0.6%(基于玉米秸秆的干重),120℃,反应45min。
2.2 LHW预处理:将粉碎后的玉米秸秆置于反应釜中,玉米秸秆的干物浓度为20%(基于总质量),200℃,反应5min。
3.将预处理后的秸秆置于酶水解反应器中,底物质量浓度为3%,加入水解酶和柠檬酸-柠檬酸钠缓冲液,在50℃的振荡箱(250rpm)内进行酶解反应,酶水解24小时。
Claims (10)
1.利用氨和助剂对木质纤维素进行预处理的方法,其特征在于,包括以下步骤:
将木质纤维素原料和助剂水溶液均匀混合,通入或加入氨,至混合基质呈碱性,氨的质量为木质纤维素干重的0.5~6.0,80℃~140℃下进行氨法预处理,所述的助剂选自蒽醌或蒽醌衍生物、绿氧、甲醇、异丙醇或甲醛。
2.根据权利要求1所述的方法,其特征在于,所述的木质纤维素选自小麦秸秆、玉米秸秆、农林废料、水稻秸秆、高粱秸秆、大豆秸秆、林业废料、回收木浆纤维、木屑、软木、硬木或动物粪便。
3.根据权利要求1所述的方法,其特征在于,所述的木质纤维素原料选自长条状木质纤维素或颗粒状木质纤维素,长条状木质纤维素的长度为0.01mm~10cm,颗粒状木质纤维素的直径为0.5mm~10cm。
4.根据权利要求1所述的方法,其特征在于,所述的木质纤维素原料在预处理过程中的质量为总质量的10%~60%。
5.根据权利要求1所述的方法,其特征在于,所述的助剂水溶液中,水为木质纤维素干重的10%~90%,助剂为木质纤维素干重的0.1~2.0%。
6.根据权利要求1所述的方法,其特征在于,所述的氨法预处理选自稀氨水预处理,氨纤维***预处理,萃取氨预处理,液氨循环预处理或液氨预处理。
7.根据权利要求1所述的方法,其特征在于,所述的氨选自氨水溶液、液氨或氨气。
8.根据权利要求1所述的方法,其特征在于,所述的氨预处理的时间为10min~360min。
9.根据权利要求1至8任一所述的方法的发酵工艺,其特征在于,包括以下步骤:
步骤1,预处理:将木质纤维素原料和助剂水溶液均匀混合,通入或加入氨,至混合基质呈碱性,氨的质量为木质纤维素干重的0.5~6.0,80℃~140℃下进行氨法预处理,所述的助剂选自蒽醌或蒽醌衍生物、绿氧、甲醇、异丙醇或甲醛;
步骤2,水解:在预处理得到的物料中加入水和水解酶,进行水解;
步骤3,发酵:加入发酵菌株,发酵,得到发酵产品。
10.根据权利要求9所述的发酵工艺,其特征在于,步骤2中,所述的水解酶选自纤维素酶和/或半纤维素酶,所述的水解酶果胶酶、木聚糖酶中的一种或者组合;步骤3中,所述的发酵菌株选自酵母、细菌或霉菌。
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