CN110140813A - 一种利用棕榈粕同时制备甘露寡糖、饲用多肽和膳食纤维的方法 - Google Patents
一种利用棕榈粕同时制备甘露寡糖、饲用多肽和膳食纤维的方法 Download PDFInfo
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Abstract
本发明属于生物技术领域，公开了一种利用棕榈粕同时制备甘露寡糖、饲用多肽和膳食纤维的方法，将棕榈粕与水按混合，添加复合酶1，酶解后离心分离，上清液干燥后获得甘露寡糖，甘露寡糖对棕榈粕中甘露聚糖的收得率≥80%，甘露糖占总糖比例≥95%。富含粗蛋白的固形物与水混合，添加复合酶2，酶解后离心分离，上清液干燥后获得饲用多肽，多肽对棕榈粕中蛋白质的得率≥90%，小肽含量≥80%。富含粗纤维的固形物添加复合酶3和复合菌剂，发酵后获得饲用膳食纤维，其中可溶性纤维含量30－50%，甘露寡糖含量10－20%，乳酸含量≥8%，乳酸菌≥5×109CFU/g，酵母菌≥1×109CFU，是一种优质的饲料添加剂。
Description
技术领域
本发明属于生物技术领域，具体涉及一种利用棕榈粕同时制备甘露寡糖、饲用多肽和膳食纤维的方法。
背景技术
棕榈粕为棕榈仁果榨油后的副产品，颜色呈棕色，有巧克力气味。棕榈仁粕中粗蛋白和粗脂肪含量比玉米、麸皮等常规饲料高，是一种性价比较高的饲料原料。棕榈仁粕中含有丰富的磷、铜、锰、钾等矿物质，以及多种维生素和氨基酸，可以作为能量饲料与其他饲料混合使用，有较好的应用潜力。
但棕榈粕在实际使用上尚存在限制：第一，由于加工条件影响，其口味略有苦涩，部分壳状碎屑质地比较坚硬，影响了禽畜的食用。第二，棕榈粕蛋白含量较低，在15％－20％，难以跟豆粕、棉粕、菜粕等蛋白原料相比而取而代之。第三，棕榈粕中含有较多的非淀粉多糖，其中直链甘露聚糖78％、阿拉伯糖基木聚糖3％、葡萄糖醛酸木聚糖3％、纤维素12％，特别是甘露聚糖有很强的抗营养作用，它会降低家禽和猪的消化率；并抑制动物胰岛素分泌，影响动物体内糖代谢和对葡萄糖、水的利用；还会引起肠道免疫系统应激反应，导致动物出现腹泻、采食量下降等不良反应。由于上述原因，棕榈粕主要局限在禽类和反刍动物中应用。
棕榈粕通过发酵和酶解，提高在饲料中应用价值的案例很多。但基本都是通过固态发酵酶解工艺，没有经过分离提纯有效成分而直接饲用。优点是工艺简单，成本较低；但存在酶解效率低，尤其是针对甘露聚糖，因此这种固态发酵酶解的棕榈粕跟原料棕榈粕相比，没有特别的改进，即大量的抗营养因子仍然存在，蛋白质也得不到提高，造成了棕榈粕资源没有有效利用。
甘露聚糖对于动物而言，是一个难以消化利用的抗营养因子，但如果酶解成为甘露寡糖或者甘露单糖，具有抗菌消炎，提高动物免疫力的功效，是一种较好的保健食品或饲料添加剂原料。目前甘露寡糖或单糖，主要来自于魔芋酶解，由于魔芋中甘露聚糖很容易分离溶出，采用单一甘露聚糖酶就容易得到甘露寡糖或单糖。但棕榈粕中甘露聚糖通常与纤维素和半纤维素结合，单一酶酶解困难，得率低。虽然王绍云等有通过单一的内切甘露聚糖酶液体酶解棕榈粕并进行甘露寡糖组分研究的报道，但需要经过氢氧化钠预处理，加酶量大(5g酶/100g棕榈粕)，得率低，成本高。
本发明利用酶解方式将棕榈粕中的甘露聚糖变成甘露寡糖，并提取分离作为食品或饲料添加剂；然后再进一步酶解其中的蛋白质，制备多肽，蛋白质含量高，作为优质饲用蛋白质原料；最后将残留的纤维类固形物通过发酵酶解方式，制备饲用膳食纤维，实现了棕榈粕的超值综合利用。
发明内容
本发明的目的是利用酶解方式将棕榈粕中的甘露聚糖变成甘露寡糖，并提取分离作为食品或饲料添加剂；同时对提取后的剩余固形物再进行酶解，制备多肽，作为优质饲用蛋白质原料；最后将残留的纤维类固形物通过发酵酶解方式，制备饲用膳食纤维。
为了达到上述目的，本发明采取以下技术措施：
一种利用棕榈粕同时制备甘露寡糖、饲用多肽和膳食纤维的方法，包括下述步骤：
1)将棕榈粕与水按质量比1：5－10混合，再按棕榈粕质量的0.1－0.5％加入复合酶1，控制温度50－60℃，pH4－5，酶解12－24h后，离心分离获得富含甘露寡糖的上清液和富含蛋白质的固形物；上清液进行纯化、浓缩、喷雾干燥获得甘露寡糖产品；
所述的复合酶1，每g所含的酶活性配比：甘露聚糖酶30000－50000μ，半乳糖甘露聚糖酶10000－20000μ，β－葡聚糖酶5000－10000μ；
2)富含蛋白质的固形物与水按质量比1：3－5混合，按照固形物干重0.1－1.0％添加复合酶2，控制温度40－50℃，pH5－6，酶解6－12h后，离心分离获得富含多肽的上清液与富含粗纤维的固形物，上清液进行纯化、浓缩、喷雾干燥获得饲用多肽产品；
所述的复合酶2，每g所含的酶活性配比：酸性蛋白酶3000－5000μ，中性蛋白酶20000－30000μ，碱性蛋白酶10000－20000μ；
3)富含粗纤维的固形物按干物质量添加2％碳酸钙，0.1－0.5％复合酶3和0.1－0.5％复合乳酸菌菌剂，控制温度30－40℃，发酵24－36h，经干燥、粉碎，并在每g产品中添加1×109CFU的酵母菌，获得饲用膳食纤维产品；
所述的复合酶3，每g所含的酶活性配比：纤维素酶10000－20000μ，木聚糖酶10000－20000μ，β－葡聚糖酶5000－10000μ，甘露聚糖酶10000－20000μ。
所述的复合乳酸菌，每克含有：乳酸片球菌5－10×109CFU，干酪乳杆菌1－5×109CFU，凝结芽胞杆菌1－2×109CFU；
以上所述的方法中，优选的，所述的乳酸片球菌的保藏编号为CICC10344，干酪乳杆菌的保藏编号为CICC20280，凝结芽胞杆菌的保藏编号为CICC23843；
以上所述的方法中，优选的，酵母菌的保藏编号为ATCC9763。
与现有技术相比，本发明具有以下优点：
1、本发明首次针对棕榈粕中的各物质含量进行分析后，配制复合酶制剂，利用复合酶的协同作用，分步酶解制备甘露寡糖和多肽，甘露寡糖对棕榈粕中甘露聚糖的收得率≥80％；多肽对棕榈粕中蛋白质的收得率≥90％；
2、本发明采用发酵酶解协同的方法利用提取甘露寡糖和多肽后的残留物制备饲用膳食纤维，产品富含纤维、甘露寡糖、乳酸以及益生菌，实现了棕榈粕原料的全面综合利用。
3.利用本发明的方法，甘露寡糖对棕榈粕中甘露聚糖的收得率≥80％，获得的甘露寡糖产品，甘露糖占总糖比例≥95％；获得的多肽对棕榈粕中蛋白质的收得率≥90％，饲用多肽中小肽含量≥80％；获得的饲用膳食纤维产品，其中可溶性纤维含量30－50％，甘露寡糖含量10－20％，乳酸含量≥8％，乳酸菌含量≥5×109CFU/g，酵母菌≥1×109CFU。
具体实施方式
下面对本发明作进一步详细说明。本发明实施例所述技术方案，如未特备说明，均为本领域的常规技术，所述试剂或材料，如未特别说明，均来源于商业渠道。
实施例1：
一种利用棕榈粕同时制备甘露寡糖、饲用多肽和膳食纤维的方法，包括下述步骤：
(1)100kg棕榈粕，甘露聚糖含量38％，粗蛋白含量17％，粉碎过40目，加水500kg，混合均匀调pH4.5，加入100g复合酶1，55℃酶解20h，5000rpm离心10min，获得上清液和富含蛋白质的固形物，上清液进行纯化，浓缩和喷雾干燥，可获得甘露寡糖制品32.5kg，相对于原料中甘露总糖得率为85.5％。
(2)多肽制备：富含蛋白质的固形物与水按质量比1：3.5混合，按干物质量的0.2％添加复合酶2，控制温度50℃，pH5.5，酶解10h后，离心分离获得富含多肽的上清液与富含粗纤维的固形物，上清液进行纯化、浓缩、喷雾干燥获得饲用多肽产品15.5kg，相对于原料中蛋白质的得率为91.2％。
(3)饲用膳食纤维制备：富含粗纤维的固形物按干物质量添加2％碳酸钙，混合均匀后再添加0.2％复合酶3和0.1％复合乳酸菌菌剂，控制温度34℃，发酵24h，经干燥、粉碎，并在每g产品中添加1×109CFU的酵母菌(ATCC9763)，获得饲用膳食纤维产品，总纤维含量62％，可溶性纤维含量41％(占总物质重)；甘露寡糖含量12.1％(占总物质重)，乳酸含量8.6％，乳酸菌含量6.5×109CFU/g，酵母菌1.2×109CFU。
所述的复合酶1包括：甘露聚糖酶50000μ，半乳糖甘露聚糖酶10000μ，β－葡聚糖酶10000μ。
复合酶2包括：酸性蛋白酶5000μ，中性蛋白酶30000μ，碱性蛋白酶20000μ。
复合酶3包括：纤维素酶20000μ，木聚糖酶20000μ，β－葡聚糖酶10000μ，甘露聚糖酶10000μ。
每克复合乳酸菌制剂含有：乳酸片球菌(CICC10344)5×109CFU，干酪乳杆菌(CICC20280)5×109CFU，凝结芽胞杆菌(CICC23843)1×109CFU。
上述实施例仅例示性说明本发明的原理及其功效，以及部分运用的实施例，对于本领域的普通技术人员来说，在不脱离本发明创造构思的前提下，还可以做出若干变形和改进，这些都属于本发明的保护范围。
Claims (4)
1.一种利用棕榈粕同时制备甘露寡糖、饲用多肽和膳食纤维的方法，包括下述步骤：
1）将棕榈粕与水按质量比1：5－10混合，再按棕榈粕质量的0.1－0.5%加入复合酶1，控制温度50－60℃，pH4－5，酶解12－24h后，离心分离获得富含甘露寡糖的上清液和富含蛋白质的固形物；上清液进行纯化、浓缩、喷雾干燥获得甘露寡糖产品；
所述的复合酶1，每g所含的酶活性配比：甘露聚糖酶30000－50000μ，半乳糖甘露聚糖酶10000－20000μ，β－葡聚糖酶5000－10000μ；
2）富含蛋白质的固形物与水按质量比1：3－5混合，按照固形物干重0.1－1.0%添加复合酶2，控制温度40－50℃，pH5－6，酶解6－12h后，离心分离获得富含多肽的上清液与富含粗纤维的固形物，上清液进行纯化、浓缩、喷雾干燥获得饲用多肽产品；
所述的复合酶2，每g所含的酶活性配比：酸性蛋白酶3000－5000μ，中性蛋白酶20000－30000μ，碱性蛋白酶10000－20000μ；
3）富含粗纤维的固形物按干物质量添加2%碳酸钙，0.1－0.5%复合酶3和0.1－0.5%复合乳酸菌菌剂，控制温度30－40℃，发酵24－36h，经干燥、粉碎，并在每g产品中添加1×109CFU的酵母菌，获得饲用膳食纤维产品；
所述的复合酶3，每g所含的酶活性配比：纤维素酶10000－20000μ，木聚糖酶10000－20000μ，β－葡聚糖酶 5000－10000μ，甘露聚糖酶10000－20000μ。
2.根据权利要求1所述的方法，所述的复合乳酸菌，每克含有：乳酸片球菌5－10×109CFU，干酪乳杆菌1－5×109CFU，凝结芽胞杆菌1－2×109CFU。
3.根据权利要求1所述的方法，所述的乳酸片球菌的保藏编号为CICC10344，干酪乳杆菌的保藏编号为CICC20280，凝结芽胞杆菌的保藏编号为CICC23843。
4.根据权利要求1所述的方法，所述的酵母菌的保藏编号为ATCC9763。
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