CN116491653A - 玉米芯提取物及其制备方法与应用 - Google Patents
玉米芯提取物及其制备方法与应用 Download PDFInfo
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
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- B01D11/0492—Applications, solvents used
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- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation or decoction
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Abstract
本发明涉及玉米芯提取物的制备方法及其在益生菌中的应用,属于功能食品技术领域。本发明提供了一种玉米芯提取物的制备方法,首先将玉米芯进行烘干、磨粉,加入水提取,将第一次固液分离所得液相进行浓缩,加入无水乙醇混合,第二次固液分离取固相,得到玉米芯水溶性提取物;再将第一次固液分离所得固相烘干,加入碱性溶液和过氧化氢的混合溶液进行加热,固液分离取液相,将所得液相浓缩,加入无水乙醇混合,固液分离取固相,得到玉米芯水不溶性提取物;最后将玉米芯水溶性提取物和水不溶性提取物混合,得到玉米芯提取物。本发明中所得玉米芯提取物不仅可以促进益生菌的生长、对益生菌酸化活性影响更大,而且对益生菌的短链脂肪酸的促进作用更强。
Description
技术领域
本发明属于功能食品技术领域,具体涉及玉米芯提取物及其制备方法与应用。
背景技术
玉米是禾本科玉蜀黍属一年生草本植物,是重要的粮食作物和饲料作物,也是全世界总产量最高的农作物,其种植面积和总产量仅次于水稻和小麦。玉米芯是玉米果穗脱去籽粒后的穗轴,主要由35%-40%的半纤维素、32%-36%的纤维素、17%-20%的木质素及1.2%-1.8%的灰分构成。
目前,我国玉米芯主要用于猪饲料、纸板、水泥板、水泥砖制作等。玉米芯中功能性成分未进行充分利用,对玉米芯中的功能性成分造成了浪费。因此通过从玉米芯中提取功能性成分并应用,剩余的玉米芯残渣可照常用于纸板,猪饲料等,提高玉米芯的利用价值。
因此,需要研究不同品种的玉米芯提取物的理化性质和益生活性的差异,以有利于广泛应用在食品和药品行业中,产生很好的社会效益和经济效益。
发明内容
为解决上述技术问题,本发明提供了玉米芯提取物及其制备方法与应用,通过对不同品种的玉米芯进行提取,制备得到了可以促进益生菌增殖、促进酸化活性和促进短链脂肪酸增加的玉米芯提取物。
本发明通过以下技术方案实现:
本发明第一个目的是提供一种玉米芯提取物的制备方法,包括以下步骤:
(1)将玉米芯进行烘干、磨粉,加入水提取,第一次固液分离得固相和液相,将所得液相进行浓缩,加入无水乙醇混合,第二次固液分离取固相,将所得固相干燥,除淀粉,得到玉米芯水溶性提取物;
(2)将步骤(1)中第一次固液分离所得固相烘干,加入碱性溶液和过氧化氢的混合溶液进行加热,固液分离取液相,将所得液相浓缩,加入无水乙醇混合,固液分离取固相,将所得固相干燥,除淀粉和除蛋白质后,得到玉米芯水不溶性提取物;
(3)将步骤(1)所得玉米芯水溶性提取物和步骤(2)所得水不溶性提取物混合,得到玉米芯提取物。
在本发明的一个实施例中,步骤(1)中,所述玉米芯选自糯玉米芯和/或甜玉米芯。
在本发明的一个实施例中,所述糯玉米芯的玉米品种选自中糯2号、申科糯602和沪红糯1号中的一种或多种;所述甜玉米芯的玉米品种选自金银208、雪甜7401和申雪甜1号中的一种或多种。
在本发明的一个实施例中,步骤(1)中,按照料液比1:15-1:20加入水。
在本发明的一个实施例中,步骤(1)中,所述提取的条件为:80℃-88℃提取2h-2.5h。
在本发明的一个实施例中,步骤(1)中,按照料液比1:3-1:4加入无水乙醇。
在本发明的一个实施例中,所述碱性溶液选自氢氧化钠溶液和/或氢氧化钾溶液。
在本发明的一个实施例中,步骤(2)中,按照料液比1:15-1:20加入碱性溶液和过氧化氢的混合溶液。
本发明第二个目的是提供所述的制备方法制备所得玉米芯提取物。
本发明第三个目的是提供所述的玉米芯提取物在益生菌中的应用。
本发明的机理:
(1)玉米芯提取物的提取机理:水溶性提取物自身可溶于水中,易被热水提取。水不溶性提取物与玉米芯中的纤维素相互缠绕、与木质素共价连接,不易被水提取。而碱性溶液可以将提取物和纤维素分开,同时过氧化氢可以氧化木质素,使水不溶性提取物被提取。
(2)在益生菌中的机理:益生菌中存在可将玉米芯提取物降解的β-木聚糖酶等酶,益生菌可利用玉米芯提取物降解后的碳源作为营养物质实现自身生长。玉米芯提取物中包含水溶性和水不溶性的提取物,在应用到益生活性时可以产生协同作用。水溶性提取物和水不溶性提取物可以相互作用可以提高益生菌中酶的活性,进而更有利于促进益生菌的生长。
本发明的上述技术方案相比现有技术具有以下优点:
本发明提供了玉米芯提取物及其制备方法与应用,通过从不同品种的玉米芯中提取玉米芯提取物,其中水溶性提取物和水不溶性提取物中均包含阿拉伯糖、木糖、半乳糖、葡萄糖、甘露糖和Ara/Xyl等成分;且本发明中混合后的玉米芯提取物不仅可以促进益生菌的生长、对益生菌酸化活性影响更大,而且对益生菌的短链脂肪酸的促进作用更强。
附图说明
为了使本发明的内容更容易被清楚的理解,下面根据本发明的具体实施例并结合附图,对本发明作进一步详细的说明,其中
图1是本发明测试例3中玉米芯提取物的扫描电镜图;
图2是本发明测试例4中玉米芯提取物对益生菌生长的影响;其中,图2-A为水溶性提取物对植物乳杆菌生长的影响;图2-B为水不溶性提取物对植物乳杆菌生长的影响;图2-C为水溶性提取物对动物双歧杆菌生长的影响;图2-D为水不溶性提取物对动物双歧杆菌生长的影响;
图3是本发明测试例4中玉米芯提取物对益生菌酸化活性的影响;其中图3-A为水溶性提取物对植物乳杆菌酸化活性的影响;图3-B为水不溶性提取物对植物乳杆菌酸化活性的影响;图3-C为水溶性提取物对动物双歧杆菌酸化活性的影响;图3-D为水不溶性提取物对动物双歧杆菌酸化活性的影响;
图4是本发明测试例5中玉米芯提取物对植物乳杆菌促进乙酸增加的影响;
图5是本发明测试例5中玉米芯提取物对植物乳杆菌促进丙酸增加的影响;
图6是本发明测试例5中玉米芯提取物对植物乳杆菌促进丁酸增加的影响;
图7是本发明测试例5中玉米芯提取物对植物乳杆菌促进乙酸增加的影响;
图8是本发明测试例5中玉米芯提取物对植物乳杆菌促进丙酸增加的影响;
图9是本发明测试例5中玉米芯提取物对植物乳杆菌促进丁酸增加的影响。
具体实施方式
下面结合附图和具体实施例对本发明作进一步说明,以使本领域的技术人员可以更好地理解本发明并能予以实施,但所举实施例不作为对本发明的限定。
实施例1
本实施例提供了一种玉米芯提取物的制备方法,具体包括以下步骤:
(1)将糯玉米芯(中糯2号ZW、申科糯602SW、沪红糯1号HW)烘干,磨粉后,按照料液比1:15(w/v),80℃热水提取2h,离心取上清液。将上清液进行浓缩,加入四倍体积无水乙醇沉淀多糖,离心后冷冻干燥。后采用反复冻融法除淀粉,即得水溶性提取物,分别标记为ZWE、SWE和HWE。
(2)将步骤(1)中第一次离心后的残渣烘干至恒重,加入0.16mol/LNaOH,1.5%H2O2(料液比1:15)的溶液中80℃加热2h,离心取上清液。将上清液浓缩后,加入四倍体积无水乙醇沉淀多糖,离心后冷冻干燥,后采用反复冻融法除淀粉,采用碱性蛋白酶(Alcalase2.4L)除蛋白质,即得水不溶性提取物,分别标记为ZWU、SWU和HWU。
(3)将步骤(1)得到的水溶性提取物与步骤(2)得到的水不溶性提取物进行混合,得到玉米芯提取物。
实施例2
本实施例提供了一种玉米芯提取物的制备方法,具体包括以下步骤:
(1)将甜玉米芯(金银208JS、雪甜7401XS、申雪甜1号SS)烘干,磨粉后,按照料液比1:15(w/v),80℃热水提取2h,离心取上清液。将上清液进行浓缩,加入四倍体积无水乙醇沉淀多糖,离心后冷冻干燥。后采用反复冻融法除淀粉,即得水溶性提取物,分别标记为JSE、XSE和SSE。
(2)将步骤(1)中第一次离心后的残渣烘干至恒重,加入0.16mol/LNaOH,1.5%H2O2(料液比1:15)的溶液中80℃加热2h,离心取上清液。将上清液浓缩后,加入四倍体积无水乙醇沉淀多糖,离心后冷冻干燥,后采用反复冻融法除淀粉,采用碱性蛋白酶(Alcalase2.4L)除蛋白质,即得水不溶性提取物,分别标记为JSU、XSU和SSU。
(3)将步骤(1)得到的水溶性提取物与步骤(2)得到的水不溶性提取物进行混合,得到玉米芯提取物。
对比例
将玉米须粉末用95%乙醇进行处理后晾干。称取玉米须粉末1kg于提取容器中,用蒸馏水(1∶20,w/v),80℃加热2h,提取2次,过滤、离心取其上清液,浓缩至原体积的1/3,加入4倍体积无水乙醇至浓缩液中,4℃静置12h,离心取沉淀物,用无水乙醇洗涤两次,收集沉淀物,真空冷冻干燥,称重并记录,计算多糖得率。
配置1%玉米须多糖溶液,加入Sevag试剂,常温条件下旋转搅拌30min,静置30min,离心去除蛋白质,重复以上步骤3次。去除蛋白质的多糖溶液加入无水乙醇至80%,4℃静置12h,离心反复冻融去除淀粉即得到玉米须多糖(CSP)。
测试例1
对实施例和对比例中不同品种玉米芯制备所得水溶性提取物和水不溶性提取物的得率和化学组成进行检测。结果如表1和表2所示:
表1水溶性提取物和水不溶性提取物的得率和化学组成
表2
n.a.表示未测出。
由表1和表2可以看出,玉米芯提取物的中性糖含量较高,糖醛酸含量在10%以下,蛋白质含量低于4%。在水溶性提取物和水不溶性提取物比较中,水溶性提取物的中性糖含量在54.94%-77.77%之间,而水不溶性提取物的中性糖含量在28.13%-51.32%之间。很明显水溶性提取物的中性糖含量高于水不溶性提取物。水不溶性提取物的糖醛酸含量高于水溶性提取物。在单糖组成方面,大部分的水溶性提取物中发现甘露糖的存在。在Ara/Xyl中,甜玉米芯提取物的Ara/Xyl高于糯玉米芯,表明甜玉米芯中的提取物中阿拉伯木聚糖的分支程度高于糯玉米。由此可以看出,甜玉米提取物中的多糖在结构特征方面可能与糯玉米的多糖有所不同。
测试例2
对实施例1中不同品种玉米芯制备所得水溶性提取物和水不溶性提取物的平均分子量进行检测,结果如表3所示:
表3玉米芯提取物的平均分子量
其中,表示重均分子量/>表示数均分子量。
由表3可见,大部分样品的多分散指数(PDI,)较小,表明玉米芯提取物样品分子量具有同质性。除沪红糯1号和申雪甜1号的玉米芯中的提取物之外,水不溶性提取物的/>大于水溶性提取物。另一方面,在水溶性提取物中,甜玉米芯水溶性提取物的/>大于糯玉米芯水溶性提取物。
测试例3
对实施例中不同品种玉米芯制备所得水溶性提取物和水不溶性提取物的微观结构进行检测,结果如图1所示:
图1为玉米芯提取物的500×的扫描电镜图。由图1可以看出,玉米芯水溶性提取物的结构成海绵网络状,其表面形态像无数海绵形成的,而水不溶性提取物呈现片状,排列无序。
测试例4
对实施例中不同品种玉米芯制备所得水溶性提取物和水不溶性提取物以及玉米芯提取物进行益生菌活性检测,结果如图2和图3所示:
图2为植物乳杆菌和动物双岐杆菌的生长曲线,以MRS培养基(购于北京索莱宝科技公司,型号M8540)为空白对照,以含有1%低聚木糖的MRS培养基为阳性对照,实验组为分别含有1%玉米芯提取物的MRS培养基。接菌后,进行厌氧发酵,每2h测定OD600。由图2可以看出,所有样品的OD600值均高于空白组,玉米芯提取物对动物双歧杆菌和植物乳杆菌的生长有显著地促进作用。水不溶性提取物对益生菌(植物乳杆菌,动物双歧杆菌)的生长促进效果强于水溶性提取物。甜玉米芯提取物的促进益生菌生长的效果弱于糯玉米芯提取物。而在水溶性提取物和水不溶性提取物混合组(ZWE+ZWU)中,两种菌的数量最多,说明其促进益生菌生长作用最好。
图3为玉米芯提取物对植物乳杆菌和动物双歧杆菌的酸化活性的影响。厌氧培养,每2h测定pH。由图3的结果发现,所有样品的发酵液的pH均低于空白组,且糯玉米芯提取物发酵液的pH低于甜玉米芯提取物,表明糯玉米芯提取物对菌种的酸化活性影响较高。水溶性提取物和水不溶性提取物混合组中,其发酵液pH最低,说明促进益生菌产生大量的有机酸。
测试例5
乙酸,丙酸,正丁酸是体外发酵形成的主要短链脂肪酸,是多糖体外酵解的典型代表产物。其中菌种会分泌更多的乙酸。本测试例中通过将植物乳杆菌和动物双岐杆菌在培养基中厌氧发酵48h,测定短链脂肪酸的含量,结果如图4-9所示。从图4-9中可得出,实验组和阳性对照组(XOS,添加1%的低聚木糖的MRS培养基)中两种益生菌产生的的乙酸,丙酸和丁酸含量均高于空白对照组,且相对玉米须多糖,玉米芯提取物(ZW、SW、HW、JS、XS和SS)促进菌种产生更多的乙酸,丙酸和丁酸。其中水溶性提取物和水不溶性提取物混合组中的乙酸,丙酸和丁酸含量最高。
其中,在菌种产生的短链脂肪酸中,乙酸含量最高,丙酸含量最少。糯玉米芯提取物产生的乙酸高于甜玉米芯提取物。同样,水溶性提取物产生的乙酸低于水不溶性提取物。而在糯玉米芯提取物发酵液和甜玉米芯提取物发酵液中没有显著性差异,同样,大部分水溶性提取物和水不溶性提取物之间也没有显著性差异。
水溶性提取物可能会和水不溶性提取物之间相互作用,提高益生菌中的酶活,进而产生更多的碳源以供益生菌利用。
显然,上述实施例仅仅是为清楚地说明所作的举例,并非对实施方式的限定。对于所属领域的普通技术人员来说,在上述说明的基础上还可以做出其它不同形式变化或变动。这里无需也无法对所有的实施方式予以穷举。而由此所引申出的显而易见的变化或变动仍处于本发明创造的保护范围之中。
Claims (10)
1.一种玉米芯提取物的制备方法,其特征在于,包括以下步骤:
(1)将玉米芯进行烘干、磨粉,加入水提取,第一次固液分离得固相和液相,将所得液相进行浓缩,加入无水乙醇混合,第二次固液分离取固相,将所得固相干燥,除淀粉,得到玉米芯水溶性提取物;
(2)将步骤(1)中第一次固液分离所得固相烘干,加入碱性溶液和过氧化氢的混合溶液进行加热,固液分离取液相,将所得液相浓缩,加入无水乙醇混合,固液分离取固相,将所得固相干燥,除淀粉和除蛋白质后,得到玉米芯水不溶性提取物;
(3)将步骤(1)所得玉米芯水溶性提取物和步骤(2)所得水不溶性提取物混合,得到玉米芯提取物。
2.根据权利要求1所述的制备方法,其特征在于,步骤(1)中,所述玉米芯选自糯玉米芯和/或甜玉米芯。
3.根据权利要求2所述的制备方法,其特征在于,所述糯玉米芯的玉米品种选自中糯2号、申科糯602和沪红糯1号中的一种或多种;所述甜玉米芯的玉米品种选自金银208、雪甜7401和申雪甜1号中的一种或多种。
4.根据权利要求1所述的制备方法,其特征在于,步骤(1)中,按照料液比1:15-1:20加入水。
5.根据权利要求1所述的制备方法,其特征在于,步骤(1)中,所述提取的条件为:80℃-88℃提取2h-2.5h。
6.根据权利要求1所述的制备方法,其特征在于,步骤(1)中,按照料液比1:3-1:4加入无水乙醇。
7.根据权利要求1所述的制备方法,其特征在于,所述碱性溶液选自氢氧化钠溶液和/或氢氧化钾溶液。
8.根据权利要求1所述的制备方法,其特征在于,步骤(2)中,按照料液比1:15-1:20加入碱性溶液和过氧化氢的混合溶液。
9.根据权利要求1-8中任一项所述的制备方法制备所得玉米芯提取物。
10.权利要求9所述的玉米芯提取物在益生菌中的应用。
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