CN105581344A - 一种含还原型谷胱甘肽的益生菌产品及其制备方法 - Google Patents
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Abstract
本发明涉及益生菌产品,公开了一种含还原型谷胱甘肽的益生菌产品,其由益生菌在增殖培养基中利用氨基酸培养基合成还原型谷胱甘肽而得;所述益生菌为植物乳杆菌、鼠李糖乳杆菌、嗜酸乳杆菌、干酪乳杆菌、长双歧杆菌和乳双歧杆菌中的至少一种;所述增殖培养基为MRS培养基和胡萝卜汁,或者MRS培养基和番茄汁。本发明还公开了该益生菌产品的制备方法。本发明通过在各种益生菌的培养液中添加氨基酸原料,使益生菌产生具有生理活性的还原型谷胱甘肽,提高益生菌的营养效果,所得到益生菌产品同时具有益生菌和谷胱甘肽的双重保健功效。
Description
技术领域
本发明涉及益生菌产品,具体是一种含还原型谷胱甘肽的益生菌产品及其制备方法。
背景技术
益生菌是一类对宿主有益的活性微生物,是定植于人体肠道、生殖系统内,能产生确切健康功效从而改善宿主微生态平衡、发挥有益作用的活性有益微生物的总称。人体、动物体内有益的细菌或真菌主要有:乳酸菌、双歧杆菌、嗜酸乳杆菌、放线菌、酵母菌等。当人体内有足够的益生菌时,人就会处于健康的状态,但是一旦体内菌群失去平衡,比如菌种间比例发生大幅变化或者超出正常数值时,那么腹泻、过敏、胃口不佳、疲倦、免疫力低等一系列病症就会随之而来,人体的健康就会亮红灯,而这时补充适当的益生菌,协助体内菌群平衡,就能让人重现健康状态。益生菌有防治腹泻、增强人体免疫力、缓解过敏作用、降低血清胆固醇等作用。
还原型谷胱甘肽广泛存在于动、植物及各种微生物体内,在人体内的生化防御体系起重要作用,具有多方面的生理功能。它的主要生理作用是能够清除掉人体内的自由基,作为体内一种重要的抗氧化剂,保护许多蛋白质和酶等分子中的巯基,是体内主要的自由基清除剂,可防止各种肝损伤。谷胱甘肽具有整合解毒作用,能与某些药物(如扑热息痛)、毒素(如自由基、碘乙酸、芥子气,铅、汞、砷等重金属)等结合,而具有整合解毒作用。谷胱甘肽可作为功能性食品的基料,广泛应用于延缓衰老、增强免疫力、抗肿瘤等功能性食品。
发明内容
本发明的目的是提供一种含还原型谷胱甘肽的益生菌产品。
本发明的另一目的是提供该益生菌产品的制备方法。
为达到上述目的之一,本发明采用以下技术方案:
一种含还原型谷胱甘肽的益生菌产品,其由益生菌在增殖培养基中利用氨基酸培养基合成还原型谷胱甘肽而得。
进一步地,所述益生菌为植物乳杆菌、鼠李糖乳杆菌、嗜酸乳杆菌、干酪乳杆菌、长双歧杆菌和乳双歧杆菌中的至少一种。
进一步地,所述增殖培养基为MRS培养基和胡萝卜汁,或者MRS培养基和番茄汁。
进一步地,所述增殖培养基的组成为:按照质量百分数,2%葡萄糖,1%蛋白胨,1%牛肉膏,0.5%酵母浸膏,0.2%磷酸氢二钾,0.5%乙酸钠,0.05%硫酸镁,0.025%硫酸锰,0.1%吐温80,0.2%柠檬酸氢二铵,10%的胡萝卜汁或番茄汁,以及余量的蒸馏水。
进一步地,所述增殖培养基的pH为6.2~6.4,在121℃湿热灭菌20min。
进一步地,所述胡萝卜汁按以下方法制备:胡萝卜和水按照质量比1:1混合榨汁,过滤除渣,所得滤液煮沸15min。
进一步地,所述番茄汁按以下方法制备:将番茄打浆,煮沸15min后过滤除渣得滤液。
进一步地,所述氨基酸培养基的组成为:按照质量百分数,0.5%谷氨酸,0.2%半胱氨酸,0.3%甘氨酸,0.5%三磷酸腺苷二钠和余量的蒸馏水。
进一步地,所述氨基酸培养基的pH为6.2~6.4,在115℃湿热灭菌20min。
前面所述益生菌产品的制备方法,在三角瓶中装入增殖培养基,接种益生菌种子液,于厌氧箱内静置培养,然后加入氨基酸培养基,在摇床上继续培养,培养完成后,发酵液经过过滤,浓缩和冷冻干燥,得到含还原型谷胱甘肽的益生菌产品。
进一步地,所述厌氧箱内静置培养的温度是35~38℃,培养时间是12~20h;所述摇床上继续培养的温度是34~36℃,培养时间是6~10h。
进一步地,所述益生菌种子液是用MRS培养基把益生菌稀释成2~5wt%浓度而得。
本发明具有以下有益效果:
本发明通过在各种益生菌的培养液中添加氨基酸原料,使益生菌产生具有生理活性的还原型谷胱甘肽,提高益生菌的营养效果,所得到益生菌产品同时具有益生菌和谷胱甘肽的双重保健功效。
本发明的工艺过程是:先用改良的MRS培养基增殖益生菌,然后加入谷氨酸、半胱氨酸、甘氨酸及三磷酸腺苷二钠,利用益生菌自身的谷胱甘肽合成酶,由三磷酸腺苷二钠提供能量,将三种氨基酸合成谷胱甘肽。
具体实施方式
下面结合具体实施例对本发明做进一步的说明:
以下实施例中所用的菌种为:植物乳杆菌CGMCCNO.1258,鼠李糖乳杆菌CNCMI-4474、嗜酸乳杆菌CNCMI-4457,干酪乳杆菌CNCMI-4458,长双歧杆菌CNCMI-4456,乳双歧杆菌CNCMI-4472。CGMCC为中国微生物菌种保藏中心,CNCM为法国巴斯德微生物菌种保藏中心。
实施例1
按照以下步骤制备益生菌产品:
(1)胡萝卜汁的制备:胡萝卜和水按照质量比1:1混合榨汁,过滤除渣,所得滤液煮沸15min;
(2)增殖培养基的制备:2g葡萄糖,1g蛋白胨,1g牛肉膏,0.5g酵母浸膏,0.2g磷酸氢二钾,0.5g乙酸钠,0.05g硫酸镁,0.025g硫酸锰,0.1g吐温80,0.2g柠檬酸氢二铵,10g胡萝卜汁,加蒸馏水至100mL,调pH为6.2~6.4,在121℃湿热灭菌20min;
(3)氨基酸培养基的制备:0.5g谷氨酸,0.2g半胱氨酸,0.3g甘氨酸,0.5g三磷酸腺苷二钠,加蒸馏水至100mL,调pH为6.2~6.4,在115℃湿热灭菌20min;
(4)在250mL三角瓶中装入75mL增殖培养基,接种3wt%的植物乳杆菌种子液(植物乳杆菌稀释于MRS培养基),于厌氧箱内37℃静置培养16h,取出,在超净工作台上加入25mL氨基酸培养基,然后在摇床上250rpm、35℃继续培养8h;统计活菌数,用高效液相法检测培养液中的还原型谷胱甘肽,结果如表1所示;
(5)培养完成后,发酵液经过过滤,浓缩和冷冻干燥,得到含还原型谷胱甘肽的益生菌产品。
实施例2
以鼠李糖乳杆菌作为益生菌制备益生菌产品,步骤与实施例1相同。
实施例3
以嗜酸乳杆菌作为益生菌制备益生菌产品,步骤与实施例1相同。
实施例4
以干酪乳杆菌作为益生菌制备益生菌产品,步骤与实施例1相同。
实施例5
以长双歧杆菌作为益生菌制备益生菌产品,步骤与实施例1相同。
实施例6
以乳双歧杆菌作为益生菌制备益生菌产品,步骤与实施例1相同。
表1
活菌数 | 还原型谷胱甘肽峰面积 | 还原型谷胱甘肽含量 | |
实施例1(植物乳杆菌) | 4.83×109cfu/mL | 7941502 | 2.82mg/mL |
实施例2(鼠李糖乳杆菌) | 4.13×109cfu/mL | 6294118 | 2.24mg/mL |
实施例3(嗜酸乳杆菌) | 3.88×109cfu/mL | 7955284 | 2.83mg/mL |
实施例4(干酪乳杆菌) | 4.16×109cfu/mL | 8776487 | 3.12mg/mL |
实施例5(长双歧杆菌) | 3.26×109cfu/mL | 6273484 | 2.23mg/mL |
实施例6(乳双歧杆菌) | 3.95×109cfu/mL | 6860129 | 2.44mg/mL |
由表1可知,采用本发明的方法所制得的益生菌产品中均含有益生菌和还原型谷胱甘肽,6种益生菌产品的活菌数均大于3.26×109cfu/ml,合成谷胱甘肽的能力以干酪乳杆菌为最好,嗜酸乳杆菌次之,长双歧杆菌最差。
实施例7
按照以下步骤制备益生菌产品:
(1)番茄汁的制备:将番茄打浆,煮沸15min后过滤除渣得滤液;
(2)增殖培养基的制备:2g葡萄糖,1g蛋白胨,1g牛肉膏,0.5g酵母浸膏,0.2g磷酸氢二钾,0.5g乙酸钠,0.05g硫酸镁,0.025g硫酸锰,0.1g吐温80,0.2g柠檬酸氢二铵,10g番茄汁,加蒸馏水至100mL,调pH为6.2~6.4,在121℃湿热灭菌20min;
(3)氨基酸培养基的制备:0.5g谷氨酸,0.2g半胱氨酸,0.3g甘氨酸,0.5g三磷酸腺苷二钠,加蒸馏水至100mL,调pH为6.2~6.4,在115℃湿热灭菌20min;
(4)在250mL三角瓶中装入75mL增殖培养基,接种5wt%的嗜酸乳杆菌种子液(嗜酸乳杆菌稀释于MRS培养基),于厌氧箱内35℃静置培养20h,取出,在超净工作台上加入25mL氨基酸培养基,然后在摇床上250rpm、34℃继续培养10h;
(5)培养完成后,发酵液经过过滤,浓缩和冷冻干燥,得到含还原型谷胱甘肽的益生菌产品。
实施例8
按照以下步骤制备益生菌产品:
(1)番茄汁的制备:将番茄打浆,煮沸15min后过滤除渣得滤液;
(2)增殖培养基的制备:2g葡萄糖,1g蛋白胨,1g牛肉膏,0.5g酵母浸膏,0.2g磷酸氢二钾,0.5g乙酸钠,0.05g硫酸镁,0.025g硫酸锰,0.1g吐温80,0.2g柠檬酸氢二铵,10g番茄汁,加蒸馏水至100mL,调pH为6.2~6.4,在121℃湿热灭菌20min;
(3)氨基酸培养基的制备:0.5g谷氨酸,0.2g半胱氨酸,0.3g甘氨酸,0.5g三磷酸腺苷二钠,加蒸馏水至100mL,调pH为6.2~6.4,在115℃湿热灭菌20min;
(4)在250mL三角瓶中装入75mL增殖培养基,接种2wt%的干酪乳杆菌种子液(干酪乳杆菌稀释于MRS培养基),于厌氧箱内38℃静置培养12h,取出,在超净工作台上加入25mL氨基酸培养基,然后在摇床上250rpm、36℃继续培养6h;
(5)培养完成后,发酵液经过过滤,浓缩和冷冻干燥,得到含还原型谷胱甘肽的益生菌产品。
以上所述,仅为本发明的具体实施方式,但本发明的保护范围并不局限于此,任何属于本技术领域的技术人员在本发明揭露的技术范围内,可轻易想到的变化或替换,都应涵盖在本发明的保护范围之内。因此,本发明的保护范围应该以权利要求的保护范围为准。
Claims (10)
1.一种含还原型谷胱甘肽的益生菌产品,其特征在于,所述益生菌产品由益生菌在增殖培养基中利用氨基酸培养基合成还原型谷胱甘肽而得。
2.根据权利要求1所述的益生菌产品,其特征在于,所述益生菌为植物乳杆菌、鼠李糖乳杆菌、嗜酸乳杆菌、干酪乳杆菌、长双歧杆菌和乳双歧杆菌中的至少一种。
3.根据权利要求1所述的益生菌产品,其特征在于,所述增殖培养基为MRS培养基和胡萝卜汁,或者MRS培养基和番茄汁。
4.根据权利要求3所述的益生菌产品,其特征在于,所述增殖培养基的组成为:按照质量百分数,2%葡萄糖,1%蛋白胨,1%牛肉膏,0.5%酵母浸膏,0.2%磷酸氢二钾,0.5%乙酸钠,0.05%硫酸镁,0.025%硫酸锰,0.1%吐温80,0.2%柠檬酸氢二铵,10%的胡萝卜汁或番茄汁,以及余量的蒸馏水。
5.根据权利要求4所述的益生菌产品,其特征在于,所述胡萝卜汁按以下方法制备:胡萝卜和水按照质量比1:1混合榨汁,过滤除渣,所得滤液煮沸15min。
6.根据权利要求4所述的益生菌产品,其特征在于,所述番茄汁按以下方法制备:将番茄打浆,煮沸15min后过滤除渣得滤液。
7.根据权利要求1所述的益生菌产品,其特征在于,所述氨基酸培养基的组成为:按照质量百分数,0.5%谷氨酸,0.2%半胱氨酸,0.3%甘氨酸,0.5%三磷酸腺苷二钠和余量的蒸馏水。
8.上述任一项权利要求所述的益生菌产品的制备方法,其特征在于,在三角瓶中装入增殖培养基,接种益生菌种子液,于厌氧箱内静置培养,然后加入氨基酸培养基,在摇床上继续培养,培养完成后,发酵液经过过滤,浓缩和冷冻干燥,得到含还原型谷胱甘肽的益生菌产品。
9.根据权利要求8所述的制备方法,其特征在于,所述厌氧箱内静置培养的温度是35~38℃,培养时间是12~20h;所述摇床上继续培养的温度是34~36℃,培养时间是6~10h。
10.根据权利要求8所述的制备方法,其特征在于,所述益生菌种子液是用MRS培养基把益生菌稀释成2~5wt%浓度而得。
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