CN106749585B - 一种丝胶蛋白的生物质提取方法及其提取物和应用 - Google Patents
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Abstract
本发明公开了一种丝胶蛋白的生物质提取方法及其提取物和应用，涉及有机物提取领域。采用壳聚糖单一组分为絮凝剂，壳聚糖乙酸溶液，在pH值为5‑6下，用壳聚糖乙酸溶液对蚕丝脱胶废水进行絮凝，回收蚕丝脱胶废水中的丝胶蛋白，工艺简单，易于操作。生成的壳聚糖‑丝胶蛋白固体，可直接应用，无需将丝胶蛋白与壳聚糖分离，无固体废物排放，不会造成二次污染。
Description
技术领域
本发明涉及有机物提取技术领域，特别涉及一种丝胶蛋白的生物质提取方法及其提取物和应用，具体为采用壳聚糖为絮凝剂对丝胶蛋白进行提取的方法，及提取出的壳聚糖-丝胶蛋白絮凝物和其应用。
背景技术
丝胶蛋白是一种高分子球状蛋白质，具有良好的护肤、调湿、保湿、抗氧化、抗菌、生物相容、可降解性能，以及促进细胞粘附与增殖等功能特性。中国作为世界上的蚕丝生产大国，每年均有数万吨的丝胶蛋白随生产废水直接排放到环境中，由于丝胶废水有机物含量严重超标，导致废水高浓度的生化需氧量(BOD)、化学需氧量(COD)和高含氮量，引起水体缺氧和富营养化，不仅严重污染环境，增加了水处理工艺复杂性和高成本，而且浪费了珍贵的天然蚕丝蛋白质资源。因此，对丝胶蛋白进行提取再利用，对于天然产物的循环再利用，实现废物资源化，具有重要意义。
目前，丝胶蛋白主要采用化学方法和物理方法进行提取。化学方法主要有化学混凝、酸沉淀和有机溶剂沉淀等方法。采用化学方法对丝胶蛋白进行提取，操作流程复杂，步骤较多，同时使用的化学试剂易给环境造成二次污染，成本较高。如专利CN201310159100.4以聚丙烯酰胺作为絮凝剂，采用喷雾干燥法得到丝胶粗粉末，再采用乙醇沉淀提纯丝胶蛋白，其优点是丝胶蛋白生物活性没有破坏，但是该方法工艺流程较多，同时使用有机溶剂及冷冻干燥装置，其成本较高。物理方法主要有离心、超滤、冰冻等方法。如专利CN201510620681.6利用透析、喷雾干燥等方法提取丝胶蛋白，其优点是所得的丝胶蛋白纯度达到90～95％，但是需要使用胰蛋白酶进行发酵，成本高。目前采用化学物理方法提取的丝胶蛋白需要进一步分离才能使用，不能一步利用。增加的操作步骤又给丝胶蛋白的提取成本带来一定压力。
发明内容
本发明的目的是提供一种丝胶蛋白的生物质提取方法及提取物和应用。采用壳聚糖为絮凝剂对丝胶蛋白进行提取，提取方法简单，无污染，提取出的壳聚糖-丝胶蛋白絮凝物为纯生物资源，不需分离即可直接应用。
本发明采用壳聚糖为絮凝剂对丝胶蛋白进行提取，该壳聚糖絮凝剂含有-NH2基团，在酸性条件下，形成-NH3 +离子，带有正电荷，在丝胶的等电点以上，可有效提取带有负电荷的丝胶蛋白。壳聚糖絮凝剂絮凝机理如下：
本发明的具体技术方案如下：
一种丝胶蛋白的生物质提取方法，包括以下步骤：
将含有壳聚糖浓度为20-60g/L的壳聚糖乙酸溶液与所述丝胶蛋白废水按体积比为2-5:10的比例混合，调节混合液的pH值为5-6，将所述混合液以500-1000rpm/min的转速搅拌10-300min后，静置至絮凝物完全沉淀，过滤，水洗，喷雾干燥。
其中，配制含有壳聚糖浓度为20-60g/L的壳聚糖乙酸溶液，具体步骤为，将壳聚糖加入到20-30g/L乙酸水溶液中，配制成20-60g/L的壳聚糖乙酸溶液。
本发明中，优选用乙酸调节混合液的pH值。所述乙酸的浓度优选为0.01-1mol/L，进一步为0.05-0.5mol/L，具体为0.1mol/L。
本发明中，混合液搅拌的速率为500-1000rpm/min，优选为700-800rpm/min；搅拌时间为10-30min，优选为5-25min，进一步为20min。
搅拌过后，将絮凝物静置至完全沉淀。优选静置时间为10-60min，进一步为20-40min，更进一步为30min。
本发明中，所述喷雾干燥的温度为70-90℃，优选为80℃。
本发明还包括采用上述方法提取出的壳聚糖-丝胶蛋白絮凝物。
本发明还包括上述壳聚糖-丝胶蛋白的应用。该壳聚糖-丝胶蛋白具有吸湿、抗氧化、抗菌等多种功能性，可直接用于化妆品、纺织品整理应用。
本发明采用壳聚糖单一组分回收蚕丝脱胶废水中的丝胶蛋白，在酸性条件下直接絮凝丝胶蛋白，工艺简单，易于操作。带正电荷的壳聚糖可快速的与带负电荷的丝胶蛋白发生凝聚，形成沉淀，处理后的蚕丝脱胶废水中丝胶蛋白的去除率可达到84％以上。生成的壳聚糖-丝胶蛋白固体，可直接应用，无需将丝胶蛋白与壳聚糖分离，无固体废物排放，不会造成二次污染。此外，丝胶中含有大量的极性氨基酸如丝氨酸、天门冬氨酸、谷氨酸等，具有良好的吸湿保湿和抗氧化功能，壳聚糖含有的氨基具有良好的吸湿保湿和抗菌功能，絮凝后的壳聚糖-丝胶蛋白可直接用于化妆品中保湿成分和抗菌成分的添加，以及纺织品功能整理，同时具有吸湿、抗氧化、抗菌等多种功能性。
具体实施方式
下面将结合本发明中的实施例，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：
(1)将壳聚糖加入到30g/L乙酸水溶液中，配成含有壳聚糖浓度为20g/L的壳聚糖乙酸溶液；
(2)取上述壳聚糖乙酸溶液20mL加入到100mL蚕丝脱胶废水中，以浓度为0.1mol/L的乙酸调节溶液的pH值为5-6，以800rpm的转速搅拌20min后，静置20min；
(3)常温过滤、水洗，90℃喷雾干燥。
实施例2：
(1)将壳聚糖加入到20g/L乙酸水溶液中，配成含有壳聚糖浓度为40g/L的壳聚糖乙酸溶液；
(2)取上述壳聚糖乙酸溶液20mL加入到80mL蚕丝脱胶废水中，以浓度为0.1mol/L的乙酸调节溶液的pH值为5-6，以600rpm的转速搅拌30min后，静置40min；
(3)常温过滤、水洗，70℃喷雾干燥。
实施例3：
(1)将壳聚糖加入到20g/L乙酸水溶液中，配成含有壳聚糖浓度为40g/L的壳聚糖乙酸溶液；
(2)取上述壳聚糖乙酸溶液20mL加入到80mL蚕丝脱胶废水中，以浓度为0.1mol/L的乙酸调节溶液的pH值为5-6，以900rpm的转速搅拌40min后，静置30min；
(3)常温过滤、水洗，80℃喷雾干燥。
实施例4：
(1)将壳聚糖加入到30g/L乙酸水溶液中，配成含有壳聚糖浓度为60g/L的壳聚糖乙酸溶液；
(2)取上述壳聚糖乙酸溶液20mL加入到50mL蚕丝脱胶废水中，以浓度为0.1mol/L的乙酸调节溶液的pH值为5-6，以1000rpm的转速搅拌50min后，静置50min；
(3)常温过滤、水洗，80℃喷雾干燥。
取桑蚕丝皂碱法脱胶液，脱胶液中丝胶蛋白浓度为1.0g/L,COD(化学需氧量)为830mg/L,按照上述实施例1-4进行丝胶蛋白絮凝，结果如表1所示：
表1实施例1-4絮凝桑蚕丝皂碱法脱胶液后丝胶蛋白、COD去除率
| 絮凝剂 | 丝胶蛋白去除率(％) | COD去除率(％) |
| 实施例1 | 78.5 | 59.8 |
| 实施例2 | 80.6 | 60.2 |
| 实施例3 | 81.2 | 61.5 |
| 实施例4 | 81.5 | 61.6 |
取柞蚕丝皂碱法脱胶液，脱胶液中丝胶蛋白浓度为0.6g/L,COD(化学需氧量)为585mg/L,按照上述实施例1-4进行丝胶蛋白絮凝，采用重铬酸钾法测定脱胶液的COD值，吸光度法测定脱胶液中丝胶蛋白浓度，结果如表2所示：
表2实施例1-4絮凝柞蚕丝皂碱法脱胶液后丝胶蛋白、COD去除率
| 絮凝剂 | 丝胶蛋白去除率(％) | COD去除率(％) |
| 实施例1 | 80.5 | 58.4 |
| 实施例2 | 81.1 | 59.7 |
| 实施例3 | 81.6 | 60.6 |
| 实施例4 | 84.7 | 61.2 |
由表1和表2可以看出，采用壳聚糖作为絮凝剂可有效回收蚕丝脱胶废水中的丝胶蛋白，丝胶蛋白去除率为80％左右，甚至可达到84.7％。工艺简单、易操作，对丝胶蛋白的絮凝效果好，成本低。整个回收过程无固体废物排放，不会造成二次污染。絮凝出的壳聚糖-丝胶蛋白固体为纯生物资源，不必分离可直接应用。丝胶中含有大量的极性氨基酸如丝氨酸、天门冬氨酸、谷氨酸等，具有良好的吸湿保湿和抗氧化功能，壳聚糖含有的氨基具有良好的吸湿保湿和抗菌功能。利用壳聚糖和丝胶蛋白所具有的功能，本发明的壳聚糖-丝胶蛋白可直接用于化妆品中保湿成分和抗菌成分的添加，以及纺织品功能整理，同时具有吸湿、抗氧化、抗菌等多种功能性。
上述实施方式旨在举例说明本发明可为本领域专业技术人员实现或使用，对上述实施方式进行修改对本领域的专业技术人员来说将是显而易见的，故本发明包括但不限于上述实施方式，任何符合本权利要求书或说明书描述，符合与本文所公开的原理和新颖性、创造性特点的方法、工艺、产品，均落入本发明的保护范围之内。
Claims (4)
1.一种丝胶蛋白的生物质提取方法，其特征在于，包括以下步骤：
将含有壳聚糖浓度为20-60g/L的壳聚糖乙酸溶液与丝胶蛋白废水按体积比为2-5:10的比例混合，用浓度为0.1mol/L的乙酸调节混合液的pH值为5-6，将所述混合液以500-1000rpm的转速搅拌10-300min后，静置至絮凝物完全沉淀，过滤，水洗，喷雾干燥；
所述壳聚糖乙酸溶液的配制具体步骤为，将壳聚糖加入到20-30g/L乙酸水溶液中，配制成20-60g/L的壳聚糖乙酸溶液。
2.根据权利要求1所述丝胶蛋白的生物质提取方法，其特征在于，所述静置时间为10-60min。
3.如权利要求1-2任意一项所述方法提取出的壳聚糖-丝胶蛋白。
4.如权利要求3所述的壳聚糖-丝胶蛋白在化妆品及纺织品中的应用。
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