CN114480527B - 一种木质纤维素原料预处理方法及应用 - Google Patents
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Abstract
本发明公开了一种木质纤维素原料预处理方法及应用,包括以下步骤:S1、用氯化胆碱、硫脲和氢氧化钠制备木质纤维素原料预处理试剂;S2、将木质纤维素原料进行粉碎、筛选,选用粒径为20~60目作为目标原料;S3、将步骤S1制备的木质纤维素原料预处理试剂与步骤S2制备的目标原料进行混合处理,于容器中进行反应,得到预处理后的混合液;S4、将步骤S3得到的预处理后的混合液进行再生、抽滤、洗涤、烘干得到预处理后的干原料。本发明采用木质纤维素原料预处理试剂实现了木质纤维素原料高效快速预处理,同时木质纤维素原料预处理试剂的可回收利用性减少了含硫物质的排放,该预处理方法更加节能、省时和环保。
Description
技术领域
本发明属于生物化工技术领域,具体涉及一种木质纤维素原料预处理方法及应用。
背景技术
由于全球环境问题的恶化和石油资源的枯竭,木质纤维素被认为是地球上可再生、环境友好且可持续的碳资源,也被认为是生产化学品、生物燃料及生物材料的潜在原料。木质纤维素是由纤维素、半纤维素和木质素这三种主要聚合物成分组成的复杂结构。纤维素是木质纤维素的重要组成部分,其高效糖化是生产化学品和能源的关键。然而,纤维素、半纤维素和木质素通过氢键和共价键紧密结合,使得生物质具有顽固的结构,这限制了酶促和微生物的可及性降解。为了有效利用木质纤维素生物质,预处理过程对于克服生物质的顽固性是必要的。目前,针对降低纤维素结晶度、增大生物质比表面积以及半纤维素和木质素的去除率等预处理工艺已有相关报道,但大多数预处理条件剧烈不温和,能耗高且周期长并且在这些预处理过程中,会伴随大量的低价值副产品产生。因此,设计开发满足综合生物炼制要求的新工艺,实现预处理过程的低能耗、低污染、低排放且高效经济地进行生物质转化利用是至关重要的。
氢氧化钠(NaOH)可以溶胀甚至溶解纤维素,但只能溶解聚合度不高的纤维素,与硫脲(TH)的组合已被证明可以改善木质纤维素在NaOH溶液中的溶解行为(Yan,L.F.,etal.,Macromolecular Bioscience,2007,7(9-10):1139-1148.)。但是,NaOH/TH组合预处理往往需要控制在较低温度下进行(<10℃),且处理时间一般在1h以上(El-Wakil,N.A.,etal.,2008,109(5):2862-2871;Zhai,R.,et al.,2016,11(4):8703-8719.)。另外,预处理试剂NaOH/TH无再回收利用,造成成本的增加。
发明内容
解决的技术问题:针对上述技术问题,本发明提供一种木质纤维素原料预处理方法及应用,能有效解决现有木质纤维素预处理方法高能耗,低效率,有污染,预处理试剂无法回收再利用等不足之处。
技术方案:一种木质纤维素原料预处理方法,包括以下步骤:
S1、用氯化胆碱、硫脲和氢氧化钠制备木质纤维素原料预处理试剂;
S2、将木质纤维素原料进行粉碎、筛选,选用粒径为20~60目作为目标原料;
S3、将步骤S1制备的木质纤维素原料预处理试剂与步骤S2制备的目标原料进行混合处理,于容器中进行反应,得到预处理后的混合液;
S4、将步骤S3得到的预处理后的混合液进行再生、抽滤、洗涤、烘干得到预处理后的干原料。
优选的,所述步骤S1的具体过程为:将氯化胆碱和硫脲以摩尔比1:2在60~90℃下混合搅拌1~4小时,得到澄清透明的低共熔溶剂,然后将低共熔溶剂、氢氧化钠与蒸馏水混合制得木质纤维素原料预处理试剂,其中低共熔溶剂为木质纤维素原料预处理试剂总质量3~15wt.%,氢氧化钠为木质纤维素原料预处理试剂总质量5~10wt.%。
优选的,所述木质纤维素原料为甘蔗渣、玉米秸秆、玉米芯或向日葵秸秆。
优选的,所述步骤S3的具体过程为:将步骤S1制备的木质纤维素原料预处理试剂与步骤S2制备的目标原料按质量比为1:10~40进行混合,在-25~75℃下反应1~240min。
优选的,所述步骤S4的具体过程为:向步骤S3得到的预处理后的混合液中加入等体积的蒸馏水再生1min,抽滤,将得到的残渣用去离子水洗涤至中性,于烘箱中烘干至恒重,得到预处理后的干原料。
一种木质纤维素原料预处理方法在制备水解糖化液中的应用。
优选的,以上述的木质纤维素原料预处理方法处理木质纤维素原料,然后预处理后的木质纤维素原料与纤维素酶、木聚糖酶在缓冲液中酶解。
进一步的,所述纤维素酶的使用量为20~50FPU/g预处理后的干原料,所述木聚糖酶的使用量为50~80CBU/g预处理后的干原料。
进一步的,所述缓冲液为pH=4.8的50mM醋酸-醋酸钠溶液,且含有终浓度为4mg/L的四环素,所述缓冲液与预处理后的木质纤维素原料按质量比20:1进行添加。
有益效果:本发明提供了一种木质纤维素原料预处理方法及应用,采用木质纤维素原料预处理试剂实现了木质纤维素原料高效快速预处理,同时木质纤维素原料预处理试剂的可回收利用性减少了含硫物质的排放,该预处理方法更加节能、省时和环保。
附图说明
图1是本发明一种木质纤维素原料预处理方法采用的预处理试剂在不同温度下预处理甘蔗渣的效果图;
图2是本发明一种木质纤维素原料预处理方法采用的预处理试剂在不同预处理时间下预处理甘蔗渣的效果图;
图3(A)是甘蔗渣预处理前后的SEM对比表征分析图;
图3(B)是甘蔗渣预处理前后的FTIR对比表征分析图;
图3(C)是甘蔗渣预处理前后的XRD对比表征分析图;
图4是预处理试剂NaOH/ChCl-TH的回收再利用效果图。
具体实施方式
下面结合附图和具体实施例对本发明作详细说明:其中葡萄糖的浓度用高效液相色谱进行定量分析,葡萄糖得率的计算公式如下:
实施例1
将氯化胆碱和硫脲以摩尔比1:2在70℃下混合搅拌3小时,得到澄清透明的低共熔溶剂ChCl-TH,将5个500mL三口烧瓶固定并分别浸于-25℃、0℃、25℃、50℃和75℃液体浴中,向每个三口烧瓶中加入14g NaOH、24g ChCl-TH和152mL蒸馏水,搅拌混合均匀后,得到预处理试剂NaOH/ChCl-TH,然后加入10g甘蔗渣进行搅拌预处理60min。结束后,加入等体积的蒸馏水再生1min,洗净后抽滤,将滤渣置于60℃恒温箱中烘干,用于后续的酶解糖化。6个10mL酶解总体系均包括pH为4.8的50mM醋酸-醋酸钠缓冲液,终浓度为4mg/L四环素,30FPU/g纤维素酶(蔚蓝生物赠送),60CBU/g木聚糖酶(蔚蓝生物赠送),分别加入0.5g未处理的甘蔗渣(UT)和5个不同温度预处理后的甘蔗渣各0.5g,在50℃下180rpm振荡进行水解反应,监测其反应进程。从图1可以看出,不同预处理温度(-25~75℃)处理的甘蔗渣的酶解糖化率均比未处理(UT)的高,说明该预处理试剂NaOH/ChCl-TH的高效性且温度的变化对其预处理甘蔗渣的影响较小,甚至在室温条件下(25℃)即可达到较高的预处理效果。
实施例2
将7个500mL三口烧瓶固定并浸于25℃水浴中,向每个三口烧瓶中加入14g NaOH、24g ChCl-TH和152mL蒸馏水,搅拌混合均匀,随后加入10g甘蔗渣并分别在预处理时间为1min、10min、20min、40min、60min、120min和240min条件下进行搅拌预处理。结束后,加入等体积的蒸馏水再生1min,洗净后抽滤,将滤渣置于60℃恒温箱中烘干,用于后续的酶解糖化。8个10mL酶解总体系分别包括未处理的甘蔗渣(UT)0.5g和7个不同预处理时间后的甘蔗渣各0.5g,均含有pH 4.8的50mM醋酸-醋酸钠缓冲液,终浓度为4mg/L四环素,30FPU/g纤维素酶(蔚蓝生物赠送),60CBU/g木聚糖酶(蔚蓝生物赠送),在50℃下180rpm振荡进行水解反应,监测其反应进程。从图2可以看出,在室温的预处理条件下,考察了不同的预处理时间(1min~240min)发现仅需搅拌处理1min后,其糖化率即可达到最高98.6%。由此可见,该预处理试剂NaOH/ChCl-TH具有高效快速预处理木质纤维素原料的能力,与传统的高温长时预处理法相比优势明显,具有良好的工业应用前景。
实施例3
对NaOH/ChCl-TH预处理甘蔗渣前后进行SEM、FTIR和XRD表征分析。
从图3(A)SEM可以看出,纤维素致密的高级结构被破坏而变得更为疏松多孔,使得纤维素表面或内部有更多的接触位点暴露出来,增加了酶的可及度,这将更有利于纤维素酶的吸附和酶解。从图3(B)FTIR可以看出预处理前后木质纤维素中关键官能团的变化,3400cm–1和2900cm–1附近的吸收峰分别是-OH和C-H基团的伸缩振动,预处理后这两个基团的吸收峰变得更宽、更强。1730cm–1附近的峰表示与半纤维素有关的羰基,预处理后峰强度减弱,说明了一部分的半纤维素被破坏,这将导致预处理后的甘蔗渣比未处理的甘蔗渣具有更高的碳水化合物含量和葡聚糖/木聚糖比率。在1630cm–1和1515cm-1附近的峰,分别归因于木质素中芳香型C=C骨架的伸缩振动和木质素中苯环的吸收峰,预处理后吸收峰消失,说明了一部分的木质素被去除。在1378cm–1附近的峰强度增加,这是由于葡聚糖和木聚糖中的CH-弯曲振动引起。在1250cm–1附近的吸收峰是木质素酯中C-O的伸缩振动,预处理后峰强度减弱。从图3(C)XRD可以看出,在16°和22°两处较明显的衍射峰属于纤维素Ⅰ型(101和002),预处理前后的结晶指数(CrI)分别为22%和25%。CrI值的升高,说明了一些非结晶组分被去除而导致纤维素的含量上升,其被酶水解的能力越强。
实施例4
将1个500mL三口烧瓶固定并浸于25℃水浴中,将14g NaOH、24g ChCl-TH和152mL蒸馏水置于三口烧瓶中并搅拌混合均匀,随后加入10g甘蔗渣搅拌预处理1min。结束后,加入等体积的蒸馏水再生1min,抽滤并收集滤液,旋蒸除水与第一次体系质量相等(190g)时,再加入10g甘蔗渣直接用于下一轮的预处理过程,将预处理试剂NaOH/ChCl-TH回收再利用4次。5个10mL酶解总体系分别包括5个批次预处理后的甘蔗渣各0.5g,均含有pH 4.8的50mM醋酸-醋酸钠缓冲液,终浓度为4mg/L四环素,30FPU/g纤维素酶(蔚蓝生物赠送),60CBU/g木聚糖酶(蔚蓝生物赠送),在50℃下180rpm振荡进行水解反应72h,检测其葡萄糖得率。如图4所示,第一批次使用的预处理试剂糖化率是98.6%,第五批次使用的预处理试剂糖化率是72.6%,第五批次与第一批次相比保持了73.6%的糖化率。重复性良好,说明了该预处理试剂NaOH/ChCl-TH具有良好的回收再利用性,使得预处理过程更经济、环保。
以上所述仅为本发明的较佳实施例而已,并不用以限制本发明,凡在本发明的精神和原则之内所作的任何修改、等同替换和改进等,均应包含在本发明的保护范围之内。
Claims (5)
1.一种木质纤维素原料预处理方法,其特征在于,包括以下步骤:
S1、用氯化胆碱、硫脲和氢氧化钠制备木质纤维素原料预处理试剂,具体过程为:将氯化胆碱和硫脲以摩尔比1:2在60~90℃下混合搅拌1~4小时,得到澄清透明的低共熔溶剂,然后将低共熔溶剂、氢氧化钠与蒸馏水混合制得木质纤维素原料预处理试剂,其中低共熔溶剂为木质纤维素原料预处理试剂总质量3~15 wt.%,氢氧化钠为木质纤维素原料预处理试剂总质量5~10wt.%;
S2、将木质纤维素原料进行粉碎、筛选,选用粒径为20~60目作为目标原料;
S3、将步骤S1制备的木质纤维素原料预处理试剂与步骤S2制备的目标原料按质量比为1:10~40进行混合处理,于容器在-25~75℃下反应1~240min,得到预处理后的混合液;
S4、将步骤S3得到的预处理后的混合液进行再生、抽滤、洗涤、烘干得到预处理后的干原料,具体过程为:向步骤S3得到的预处理后的混合液中加入等体积的蒸馏水再生1min,抽滤,将得到的残渣用去离子水洗涤至中性,于烘箱中烘干至恒重,得到预处理后的干原料;
所述木质纤维素原料为甘蔗渣。
2.权利要求1所述的木质纤维素原料预处理方法在制备水解糖化液中的应用。
3.一种木质纤维素原料酶解的方法,其特征在于:以权利要求1-2任一项所述的木质纤维素原料预处理方法处理木质纤维素原料,然后预处理后的木质纤维素原料与纤维素酶、木聚糖酶在缓冲液中酶解。
4. 根据权利要求3所述的一种木质纤维素原料酶解的方法,其特征在于:所述纤维素酶的使用量为20~50 FPU/g预处理后的干原料,所述木聚糖酶的使用量为50~80 CBU/g预处理后的干原料。
5. 根据权利要求3所述的一种木质纤维素原料酶解的方法,其特征在于:所述缓冲液为pH=4.8的50 mM醋酸-醋酸钠溶液,且含有终浓度为4mg/L的四环素,所述缓冲液与预处理后的木质纤维素原料按质量比20:1进行添加。
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