CN111567711A - 一种具有高生物可利用度的黄嘌呤氧化酶抑制活性酵素饮料的制备方法 - Google Patents
一种具有高生物可利用度的黄嘌呤氧化酶抑制活性酵素饮料的制备方法 Download PDFInfo
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- CN111567711A CN111567711A CN202010400155.XA CN202010400155A CN111567711A CN 111567711 A CN111567711 A CN 111567711A CN 202010400155 A CN202010400155 A CN 202010400155A CN 111567711 A CN111567711 A CN 111567711A
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
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- A—HUMAN NECESSITIES
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Abstract
本发明公开了一种具有高生物可利用度的黄嘌呤氧化酶抑制活性酵素饮料的制备方法,具体是将橙汁、菊花汁、紫薯泥、茉莉花按照一定比例混合,依次通过醋酸菌、植物乳杆菌发酵后获得二次发酵酵素液,再将二次发酵酵素液进行灭菌,灭菌的二次酵素液加入多种辅料后制成酵素产品。本发明方法能够显著提高酵素高生物可利用的黄嘌呤氧化酶抑制活性,同时添加益生菌和富含膳食纤维的紫薯,这些两物质被认为具有较好的调控肠道菌群的作用。
Description
技术领域
本发明属于发酵食品制备技术领域,具体涉及一种具有高生物可利用度的黄嘌呤氧化酶抑制活性酵素饮料的制备方法。
技术背景
酵素食品是以一种或多种新鲜水果、蔬菜、菌菇、中草药等为原料,经多种微生物发酵而成的,含有丰富的小分子功能性物质、多糖及其它次生代谢产物等成分的功能性发酵产品。酵素具有多种生物功效(包括黄嘌呤氧化酶抑制活性、抗氧化活性、增强免疫力、促进消化等),追求高的保健功效是人们对产品的要求。
生物可利用是指从食物体系中释放出来,能被人体消化吸收的那部分物质,其中包含一定量的生物活性物质。生物可利用是生物活性对人体产生健康功效的前提,而生物活性物质在加工(如原料、菌种)和消化过程中会发生一系列变化,还能与加工过程中其他食品配料发生复杂的相互作用,从而对生物物质的生物可利用和健康功效产生重大影响,影响生物的健康。
近年来,随着人们生活水平的提高以及生活方式的改变,嘌呤类食物摄入量不断增加,高尿酸血症/痛风的患者也逐年增加,黄嘌呤氧化酶是形成痛风和高尿酸血症的一个关键因素,也是治疗高尿酸血症的重要靶标。非嘌呤类前体物质在体内经过一系列的转化生成嘌呤类核苷酸,继续分解生成次黄嘌呤和黄嘌呤,最终经过黄嘌呤氧化酶的连续氧化而生成尿酸。因此,黄嘌呤氧化酶直接调控着体内尿酸水平的高低。筛选具有高的黄嘌呤抑制活性被认为具有很好的治疗黄嘌呤氧化酶抑制活性。为此,本发明专利提供一项能显著提高酵素生物可利用的黄嘌呤氧化酶抑制活性的技术方案。
发明内容
本发明旨在提供一种具有高生物可利用度的黄嘌呤氧化酶抑制活性酵素饮料的制备方法。本发明方法能够显著提高生物可利用的黄嘌呤氧化酶抑制活性,具有较好的保健功效。
本发明具有高生物可利用度的黄嘌呤氧化酶抑制活性酵素饮料的制备方法,包括如下步骤:
步骤1:取甜橙(总酚含量≥0.01%)作为原料,将甜橙表皮洗净,然后在臭氧水中进行过流式浸没杀菌5~15min,沥干后依次去皮、打汁,获得甜橙汁;
步骤2:取菊花(总酚含量≥0.01%)作为原料,将菊花表皮洗净,然后在臭氧水中进行过流式浸没杀菌5~15min,沥干后,按照干物质和水质量比为1~2:4~5的比例混合,在60~80℃的水中进行加热提取2~5h,获得菊花汁;
步骤3:紫薯在110~120℃蒸煮15~20min,然后制备为紫薯泥,含水率控制在50~70%;
步骤4:醋酸菌发酵液的制备
4a、斜面菌种培养:将醋酸菌(购于济宁玉园生物科技有限公司)两环接入培养于斜面培养基中,在28~30℃下培养3~5d;所述斜面培养基的配方为:葡萄糖10g/L,酵母膏10g/L,碳酸钙10g/L,琼脂25g/L,用水定容为1L。
4b、液体菌种的制备:将斜面菌种两环直接转接至液体种子培养基中,在28~32℃下静置培养2~4d,制备得到种子液;所述液体种子培养基的配方为:葡萄糖10g/L,酵母膏10g/L,碳酸钙10g/L,用水定容为1L。
4c、扩大培养:将4b所得种子液按体积比10%的接种量转接至扩大培养基中,28~32℃下静置培养1~3d,得到醋酸菌发酵液;所述扩大培养基的配方为:葡萄糖10g/L,酵母膏10g/L,碳酸钙10g/L,用水定容为1L。
步骤5:植物乳杆菌发酵液的制备
5a、斜面菌种培养:将植物乳杆菌(购于台湾亚芯生物科技有限公司)培养于MRS斜面培养基中,在30~37℃下培养3~5d;
5b、液体菌种的制备:将斜面菌种直接转接至液体MRS种子培养基中,在30~37℃下静置培养2~4d,制备得到种子液;
5c、扩大培养:将所得种子液按10%的接种量转接至扩大培养基中,30~37℃下静置培养1-3d,得到植物乳杆菌发酵液;
步骤5中,所述MRS斜面培养基的配方构成如下:蛋白陈10.0g,牛肉粉5.0g,酵母粉4.0g,葡萄糖20.0g,吐温80 1.0mL,磷酸氢二钾2.0g,乙酸钠5.0g,柠檬酸三铵2.0g,硫酸镁0.2g,硫酸锰0.05g,琼脂粉15.0g,蒸馏水1000mL。将上述原料按配比量混合后加热溶解,校正pH值6.2,然后于121℃下高压灭菌15-20min即可。
步骤5中,所述MRS种子培养基的配方构成如下:蛋白陈10.0g,牛肉粉5.0g,酵母粉4.0g,葡萄糖20.0g,吐温80 1.0mL,磷酸氢二钾2.0g,乙酸钠5.0g,柠檬酸三铵2.0g,硫酸镁0.2g,硫酸锰0.05g,琼脂粉15.0g,蒸馏水1000mL。将上述原料按配比量混合后加热溶解,校正pH值6.2,然后于121℃下高压灭菌15-20min。
步骤5中,所述扩大培养基的配方构成如下:蛋白陈10.0g,牛肉粉5.0g,酵母粉4.0g,葡萄糖20.0g,吐温80 1.0mL,磷酸氢二钾2.0g,乙酸钠5.0g,柠檬酸三铵2.0g,硫酸镁0.2g,硫酸锰0.05g,琼脂粉15.0g,蒸馏水1000mL。将上述原料按配比量混合后加热溶解,校正pH值6.2,然后于121℃下高压灭菌15-20min。
步骤6:灭菌和发酵
6a、取甜橙汁、菊花汁、紫薯泥和茉莉花混合,在70~80℃下灭菌15~20min,冷却至室温作为发酵底物;按10:1的质量比添加醋酸菌发酵液,在28~32℃下静置发酵3~5d,过滤后即得一次发酵酵素液;
6b、将一次发酵酵素液pH值调至5.0~7.0,按10:1的质量比添加植物乳杆菌发酵液,在30~37℃下静置发酵4~5d,过滤后即得二次发酵酵素液;
步骤6a中,甜橙汁、菊花汁、紫薯泥和茉莉花混合时的质量比为5~8:1~2:2~3:0.5~1.0。
步骤7:调配
将二次发酵酵素液:木糖醇:柠檬酸:低聚果糖:山梨酸钾按照质量比5~8:0.5~0.8:0.2~0.4:0.1~0.3:0.01~0.04的比例混合调配,即得具有高生物可利用度的黄嘌呤氧化酶抑制活性酵素产品。
本发明的有益效果体现在:
1、本发明方法能够显著提高酵素可利用的黄嘌呤氧化酶抑制活性,具有较好的保健功效。
2、本发明原料主要是橙汁、菊花、紫薯,其价格便宜,开发为酵素具有较高的附加值。
3、本发明方法中添加大量的益生元和膳食纤维,具有较好的调节肠道菌群功能,且口感风味优质。
具体实施方式
黄嘌呤氧化酶抑制活性按照下表进行试验测定,依次在试管中加入溶液进行试验,于温度25℃反应30min,在290nm处测定各吸光值,各浓度组平行3次,测得平均值。
抑制率公式为:K(%)=[1-A1/A0]*100%。公式中,A1-以背景对照管作为空白对照的抑制剂管吸光度值;A0-以空白对照管作为空白对照的空白管吸光度值。
模拟体外消化和透析方法:消化过程分为两个阶段。第一阶段胃液消化。称取10.0g酵素产品,置于50mL离心管内,加入去离子水30mL,高速匀浆3min后,用6.0mol·L- 1HCl将混合液pH调至2.0,混匀,然后加入胃蛋白酶(12000U),混匀,置于37℃恒温水浴摇床中120r/min振摇2h,快速冰浴冷却后在4℃离心(10000r/min)10min,取2mL上清液作为胃液消化的样品储存在-80℃。第二阶段肠消化。用1.0mol·L-1Na2CO3溶液将胃液消化后的混合液pH调节至7.5,然后加入胰蛋白酶(1mL,4g·L-1)和胆盐(1mL,25g·L-1),混匀,置于37℃恒温水浴摇床中120r/min振摇2h。消化结束后,立即取出,冰浴使其冷却获得消化样品。模拟小肠吸收用填充纤维素透析膜胃肠消化样品,在37±2℃,以90转/分的速度持续8小时,利用能够透析的部分测定黄嘌呤氧化酶抑制活性。
以下结合具体实施例对本发明技术方案进行详细地说明。
实例1:
1、取甜橙(总酚含量≥0.01%)作为原料,将甜橙表皮洗净,然后在臭氧水中进行过流式浸没杀菌5min,沥干后依次去皮、打汁,获得甜橙汁。
2、取菊花(总酚含量≥0.01%)作为原料,将菊花表皮洗净,然后在臭氧水中进行过流式浸没杀菌5min,沥干后,按照干物质和水份数比为1:4混合,在60℃的水中进行加热提取2h,获得菊花汁。
3、紫薯在110℃蒸煮15min,然后制备为紫薯泥,含水控制在50%。
4、将醋酸菌两环接入培养于斜面培养基中,斜面培养基为葡萄糖10g/L,酵母膏10g/L,碳酸钙10g/L,琼脂25g/L,用水定容为1L。在28℃下培养3d。将斜面菌种两环直接转接至液体种子培养基中,种子培养基为葡萄糖10g/L,酵母膏10g/L,碳酸钙10g/L,用水定容为1L。在28℃下静置培养2d,制备得到种子液。将所得种子液按体积比10%的接种量转接至扩大培养基中,28℃下静置培养1d,得到醋酸菌发酵液。
5、将植物乳杆菌培养于MRS斜面培养基中,在30℃下培养3d;将斜面菌种直接转接至液体MRS种子培养基中,在30℃下静置培养2d,制备得到种子液。将所得种子液按10%的接种量转接至扩大培养基中,30℃下静置培养1d,得到植物乳杆菌发酵液。
6、取甜橙汁、菊花汁、紫薯泥、茉莉花,按照质量份数比5:1:2:0.5进行混合配制,在70℃灭菌15min,冷却至室温作为发酵底物。按10:1的质量比添加醋酸菌发酵液,在28℃下静置发酵3d,过滤后即得一次发酵酵素液。将一次发酵酵素液pH调到5.0,按10:1的质量比添加植物乳杆菌发酵液,在30℃下静置发酵4d,过滤后即得高生物可利用性黄嘌呤氧化酶抑制活性的二次发酵酵素液。
7、将二次发酵酵素液:木糖醇:柠檬酸:低聚果糖:山梨酸钾按照质量份数比为5:0.5:0.2:0.1:0.01混合调配,即得酵素产品。
分别按照表1进行试验,即将等量橙汁分别替代为不同水果汁,将植物乳杆菌发酵菌种替代为不同的乳酸菌,将低聚果糖替代为不同的低聚糖,其它步骤一致,测定酵素产品的生物可利用物的黄嘌呤氧化酶抑制活性。从表1可以看出,实例1处理较其它处理具有最高的生物可利用的黄嘌呤氧化酶抑制活性。
表1 不同处理与本方案的生物可利用黄嘌呤氧化酶抑制活性的比较
实例2:
1、取甜橙(总酚含量≥0.01%)作为原料,将甜橙表皮洗净,然后在臭氧水中进行过流式浸没杀菌10min,沥干后依次去皮、打汁,获得甜橙汁。
2、取菊花(总酚含量≥0.01%)作为原料,将菊花表皮洗净,然后在臭氧水中进行过流式浸没杀菌10min,沥干后,按照干物质和水份数比为1.5:5.5混合,在70℃的水中进行加热提取4h,获得菊花汁。
3、紫薯在115℃蒸煮17min,然后制备为紫薯泥,含水控制在60%。
4、将醋酸菌两环接入培养于斜面培养基中,斜面培养基为葡萄糖10g/L,酵母膏10g/L,碳酸钙10g/L,琼脂25g/L,用水定容为1L。在30℃下培养4d。将斜面菌种两环直接转接至液体种子培养基中,种子培养基为葡萄糖10g/L,酵母膏10g/L,碳酸钙10g/L,用水定容为1L。在32℃下静置培养3d,制备得到种子液。将所得种子液按体积比10%的接种量转接至扩大培养基中,32℃下静置培养2d,得到醋酸菌发酵液。
5、将植物乳杆菌培养于MRS斜面培养基中,在37℃下培养5d。将斜面菌种直接转接至液体MRS种子培养基中,在37℃下静置培养4d,制备得到种子液。将所得种子液按10%的接种量转接至扩大培养基中,37℃下静置培养2d,得到植物乳杆菌发酵液。
6、取甜橙汁、菊花汁、紫薯泥、茉莉花,按照质量份数比7:1.5:3:1.0进行混合配制,在75℃灭菌20min,冷却至室温作为发酵底物。按10:1的质量比添加醋酸菌发酵液,在32℃下静置发酵4d,过滤后即得一次发酵酵素液。将一次发酵酵素液pH调到7.0,按10:1的质量比添加植物乳杆菌发酵液,在37℃下静置发酵5d,过滤后即得高生物可利用性黄嘌呤氧化酶抑制活性的二次发酵酵素液。
7、将二次发酵酵素液:木糖醇:柠檬酸:低聚果糖:山梨酸钾按照质量份数比为8:0.8:0.2:0.1:0.01混合调配,即得酵素产品。
分别按照表2进行试验,即将等量橙汁分别替代为不同水果汁,将植物乳杆菌发酵菌种替代为不同的乳酸菌,将低聚果糖替代为不同的低聚糖,其它步骤一致,测定酵素产品的生物可利用物的黄嘌呤氧化酶抑制活性。从表2可以看出,实例1处理较其它处理具有最高的生物可利用的黄嘌呤氧化酶抑制活性。
表2 不同处理与本方案的生物可利用黄嘌呤氧化酶抑制活性的比较
实例3:
1、取甜橙(总酚含量≥0.01%)作为原料,将甜橙表皮洗净,然后在臭氧水中进行过流式浸没杀菌15min,沥干后依次去皮、打汁,获得甜橙汁。
2、取菊花(总酚含量≥0.01%)作为原料,将菊花表皮洗净,然后在臭氧水中进行过流式浸没杀菌5~15min,沥干后,按照干物质和水份数比为2:5混合,在80℃的水中进行加热提取5h,获得菊花汁。
3、紫薯在120℃蒸煮20min,然后制备为紫薯泥,含水控制在70%。
4、将醋酸菌两环接入培养于斜面培养基中,斜面培养基为葡萄糖10g/L,酵母膏10g/L,碳酸钙10g/L,琼脂25g/L,用水定容为1L。在30℃下培养5d。将斜面菌种两环直接转接至液体种子培养基中,种子培养基为葡萄糖10g/L,酵母膏10g/L,碳酸钙10g/L,用水定容为1L。在32℃下静置培养4d,制备得到种子液。将所得种子液按体积比10%的接种量转接至扩大培养基中,32℃下静置培养3d,得到醋酸菌发酵液。
5、将植物乳杆菌培养于MRS斜面培养基中,在37℃下培养5d。将斜面菌种直接转接至液体MRS种子培养基中,在37℃下静置培养4d,制备得到种子液。将所得种子液按10%的接种量转接至扩大培养基中,37℃下静置培养3d,得到植物乳杆菌发酵液。
6、取甜橙汁、菊花汁、紫薯泥、茉莉花,按照质量份数比8:2:3:1.0进行混合配制,在80℃灭菌20min,冷却至室温作为发酵底物;按10:1的质量比添加醋酸菌发酵液,在32℃下静置发酵5d,过滤后即得一次发酵酵素液。将一次发酵酵素液pH调到7.0,按10:1的质量比添加植物乳杆菌发酵液,在37℃下静置发酵5d,过滤后即得高生物可利用性黄嘌呤氧化酶抑制活性的二次发酵酵素液。
7、将二次发酵酵素液:木糖醇:柠檬酸:低聚果糖:山梨酸钾按照质量份数比为8:0.8:0.4:0.3:0.04混合调配,即得酵素产品。
分别按照表3进行试验,即将等量橙汁分别替代为不同水果汁,将植物乳杆菌发酵菌种替代为不同的乳酸菌,将低聚果糖替代为不同的低聚糖,其它步骤一致,测定酵素产品的生物可利用物的黄嘌呤氧化酶抑制活性。从表3可以看出,实例3处理较其它处理具有最高的生物可利用的黄嘌呤氧化酶抑制活性。
表3 不同处理与本方案的生物可利用黄嘌呤氧化酶抑制活性的比较
Claims (5)
1.一种具有高生物可利用度的黄嘌呤氧化酶抑制活性酵素饮料的制备方法,其特征在于包括如下步骤:
步骤1:取甜橙作为原料,将甜橙表皮洗净,然后在臭氧水中进行过流式浸没杀菌5~15min,沥干后依次去皮、打汁,获得甜橙汁;
步骤2:取菊花作为原料,将菊花表皮洗净,然后在臭氧水中进行过流式浸没杀菌5~15min,沥干后,按照干物质和水质量比为1~2:4~5的比例混合,在60~80℃的水中进行加热提取2~5h,获得菊花汁;
步骤3:紫薯在110~120℃蒸煮15~20min,然后制备为紫薯泥,含水率控制在50~70%;
步骤4:醋酸菌发酵液的制备
4a、斜面菌种培养:将醋酸菌两环接入培养于斜面培养基中,在28~30℃下培养3~5d;
4b、液体菌种的制备:将斜面菌种两环直接转接至液体种子培养基中,在28~32℃下静置培养2~4d,制备得到种子液;
4c、扩大培养:将4b所得种子液按体积比10%的接种量转接至扩大培养基中,28~32℃下静置培养1~3d,得到醋酸菌发酵液;
步骤5:植物乳杆菌发酵液的制备
5a、斜面菌种培养:将植物乳杆菌培养于MRS斜面培养基中,在30~37℃下培养3~5d;
5b、液体菌种的制备:将斜面菌种直接转接至液体MRS种子培养基中,在30~37℃下静置培养2~4d,制备得到种子液;
5c、扩大培养:将所得种子液按10%的接种量转接至扩大培养基中,30~37℃下静置培养1-3d,得到植物乳杆菌发酵液;
步骤6:灭菌和发酵
6a、取甜橙汁、菊花汁、紫薯泥和茉莉花混合,在70~80℃下灭菌15~20min,冷却至室温作为发酵底物,然后添加醋酸菌发酵液,发酵并过滤后即得一次发酵酵素液;
6b、将一次发酵酵素液pH值调至5.0~7.0,然后添加植物乳杆菌发酵液,发酵并过滤后即得二次发酵酵素液;
步骤7:调配
将二次发酵酵素液、木糖醇、柠檬酸、低聚果糖和山梨酸钾混合调配,即得具有高生物可利用度的黄嘌呤氧化酶抑制活性酵素产品。
2.根据权利要求1所述的制备方法,其特征在于:
步骤6a中,甜橙汁、菊花汁、紫薯泥和茉莉花混合时的质量比为5~8:1~2:2~3:0.5~1.0。
3.根据权利要求1所述的制备方法,其特征在于:
步骤6a中,按10:1的质量比添加醋酸菌发酵液,在28~32℃下静置发酵3~5d。
4.根据权利要求1所述的制备方法,其特征在于:
步骤6b中,按10:1的质量比添加植物乳杆菌发酵液,在30~37℃下静置发酵4~5d。
5.根据权利要求1所述的制备方法,其特征在于:
步骤7中,将二次发酵酵素液、木糖醇、柠檬酸、低聚果糖和山梨酸钾按照质量比5~8:0.5~0.8:0.2~0.4:0.1~0.3:0.01~0.04的比例混合调配。
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