CN108523109B - 金刺梨发酵方法 - Google Patents
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Abstract
一种金刺梨发酵方法,包括:提供一菌株混合物,该菌株混合物包括菌数比为0.3~1:0.2~0.9:1~1.8的一干酪乳酸杆菌菌株的菌液、一长双歧杆菌菌株的菌液及一酿酒酵母菌菌株的菌液;提供一金刺梨样品;及以该菌株混合物发酵该金刺梨样品,使该干酪乳酸杆菌菌株的菌液、该长双歧杆菌菌株的菌液及该酿酒酵母菌菌株的菌液为1.5×107CFU/ml~3.5×107CFU/ml的总最终浓度下,于22~33℃的温度发酵该金刺梨样品6~15天,以获得一金刺梨发酵液。借此,能够获得提升抗氧化能力的该金刺梨发酵液,以作为抗氧化保健食品食用或作为食品添加物使用,达到食用者养生的功效。
Description
技术领域
本发明关于一种金刺梨发酵方法,特别是一种获得提升抗氧化能力的金刺梨发酵液的金刺梨发酵方法。
背景技术
随着工业化的进步,我们生活的环境也充斥着越来越多的污染源,人们通过每天接触这些污染源,身体里会有许多的自由基产生,而这些自由基为引起许多现代文明病的主因。因此,现在人越来越重视抗氧化以去除自由基的保健方法。
金刺梨(Rosa sterilis var. leioclada)又称光枝无子刺梨或无籽刺梨,属蔷薇科,虽然金刺梨含有抗氧化成分,然而,必须食用大量的金刺梨才能发挥足够的保健功效,以致于效益不高。有鉴于此,需要一种金刺梨发酵方法,以获得提升抗氧化能力的金刺梨发酵液以供食用者食用,以提升保健效益。
发明内容
本发明提供一种金刺梨发酵方法,以获得提升抗氧化能力的一金刺梨发酵液。
一种金刺梨发酵方法,包括:提供一菌株混合物,该菌株混合物包括菌数比为0.3~1:0.2~0.9:1~1.8的一干酪乳酸杆菌菌株的菌液、一长双歧杆菌菌株的菌液及一酿酒酵母菌菌株的菌液;提供一金刺梨样品;及以该菌株混合物发酵该金刺梨样品,使该干酪乳酸杆菌菌株的菌液、该长双歧杆菌菌株的菌液及该酿酒酵母菌菌株的菌液为1.5×107CFU/ml~3.5×107CFU/ml的总最终浓度下,于22~33℃的温度发酵该金刺梨样品6~15天,以获得一金刺梨发酵液。
借此,以包括该干酪乳酸杆菌菌株、该长双歧杆菌菌株及该酿酒酵母菌菌株的菌株混合物,发酵该金刺梨样品,使该金刺梨样品的抗氧化活性及所含的抗氧化物质含量得以提升,以获得提升抗氧化能力的该金刺梨发酵液,能够作为抗氧化保健食品食用或作为食品添加物使用,达到食用者养生的功效。
其中,另外包括以一干酪乳酸杆菌菌株BCRC 10697、一长双歧杆菌菌株BCRC14604及一酿酒酵母菌菌株BCRC 20579分别形成该干酪乳酸杆菌菌株的菌液、该长双歧杆菌菌株的菌液及该酿酒酵母菌菌株的菌液。借此,能够获得提升抗氧化能力的该金刺梨发酵液,以作为抗氧化保健食品食用,达到食用者养生的功效。
其中,该菌株混合物于28℃的温度下,发酵该金刺梨样品。借此,能够获得提升抗氧化能力的该金刺梨发酵液,以作为抗氧化保健食品食用,达到食用者养生的功效。
其中,以该菌株混合物发酵该金刺梨样品14天。借此,能够获得提升抗氧化能力的该金刺梨发酵液,以作为抗氧化保健食品食用,达到食用者养生的功效。
附图说明
图1a:第A1~A3组的ABTS+清除率的长条图;
图1b:第A1~A3组的SOD活性的长条图;
图2a:第A1~A3组的维生素C含量的长条图;
图2b:第A1~A3组的类胡萝卜素含量的长条图;
图3a:第B1~B3组的ABTS+清除率的长条图;
图3b:第B1~B3组的SOD活性的长条图;
图3c:第B1~B3组的维生素C含量的长条图;
图3d:第B1~B3组的类胡萝卜素含量的长条图;
图4a:第C1~C3组的ABTS+清除率的长条图;
图4b:第C1~C3组的SOD活性的长条图;
图4c:第C1~C3组的维生素C含量的长条图;
图4d:第C1~C3组的类胡萝卜素含量的长条图;
图5a:第D1~D3组的ABTS+清除率的长条图;
图5b:第D1~D3组的SOD活性的长条图;
图5c:第D1~D3组的维生素C含量的长条图;
图5d:第D1~D3组的类胡萝卜素含量的长条图。
具体实施方式
为使本发明的上述及其他目的、特征及优点能更明显易懂,下文特根据本发明的实施例,并配合所附附图,作详细说明如下:
本发明提供一种金刺梨发酵方法,包括:提供一菌株混合物及一金刺梨样品,该菌株混合物包括:一干酪乳酸杆菌(Lactobacillus casei)菌株、一长双歧杆菌(Bifidobacterium longum)菌株及一酿酒酵母菌(Saccharomyces cerevisiae)菌株。以该菌株混合物发酵该金刺梨样品,即能够获得提高抗氧化能力的一金刺梨发酵液。
详言之,能够使用购自食品工业发展研究所的一干酪乳酸杆菌菌株(BCRC10697)、一长双歧杆菌菌株(BCRC14604)及一酿酒酵母菌菌株(BCRC20579)以形成该菌株混合物。
该金刺梨样品能够为一个整颗的金刺梨,例如能够将该金刺梨蒸熟以软化该金刺梨,使该菌株混合物的菌液能够渗入以利于充分发酵该金刺梨样品,或者能够将该金刺梨经过切割处理后作为该金刺梨样品以进行发酵,举例而言,能够于该金刺梨表面切割出一个切口、或将该金刺梨剖切,或者将该金刺梨切成片状等,以裸露出该金刺梨的果肉并增加与菌液接触的表面积,方便充分发酵该金刺梨。本实施例中,将该金刺梨进行榨汁形成一金刺梨汁,以作为该金刺梨样品来进行发酵,借此,相较于发酵包括果肉及果皮的整颗该金刺梨,以该金刺梨汁作为该金刺梨样品来进行发酵,能够使该菌株混合物的菌液与该金刺梨样品充分混合,进而提升发酵效率。
该菌株混合物发酵该金刺梨样品前,较佳能够先进行该干酪乳酸杆菌菌株、该长双歧杆菌菌株及该酿酒酵母菌菌株的增殖培养,活化该干酪乳酸杆菌菌株、该长双歧杆菌菌株及该酿酒酵母菌菌株,以获得健康状态的菌株,借此提高发酵的效率。本实施例中,分别将该干酪乳酸杆菌菌株、该长双歧杆菌菌株及该酿酒酵母菌菌株置于一液态培养基中进行增殖培养,以避免各菌株互相拮抗影响彼此的增殖生长,并培养至对数生长期(logphase)后分别形成一干酪乳酸杆菌菌株的菌液,一长双歧杆菌菌株的菌液及一酿酒酵母菌菌株的菌液,其中,能够分别将该干酪乳酸杆菌菌株、该长双歧杆菌菌株及该酿酒酵母菌菌株于37℃中进行增殖培养,使该干酪乳酸杆菌菌株、该长双歧杆菌菌株及该酿酒酵母菌菌株能够于最适生长温度下进行增殖,以确保菌株的品质。另外以血球细胞记数法计算菌量,分别调整该干酪乳酸杆菌菌株的菌液、该长双歧杆菌菌株的菌液及该酿酒酵母菌菌株的菌液的浓度为1×107CFU/ml,以形成用以发酵该金刺梨样品的菌株混合物,值得注意的是,以0.3~1:0.2~0.9:1~1.8的菌数比,混合该干酪乳酸杆菌菌株的菌液、该长双歧杆菌菌株的菌液及该酿酒酵母菌菌株的菌液,以形成该菌株混合物,使该干酪乳酸杆菌菌株、该长双歧杆菌菌株及该酿酒酵母菌菌株彼此之间,能够具有良好的交互作用效率,较佳能够以0.8:0.8:1.6的菌数比,混合该干酪乳酸杆菌菌株的菌液、该长双歧杆菌菌株的菌液及该酿酒酵母菌菌株的菌液,以形成该菌株混合物,进而达到最佳发酵效率。
混合该菌株混合物及该金刺梨样品,并使该干酪乳酸杆菌菌株的菌液、该长双歧杆菌菌株的菌液及该酿酒酵母菌菌株的菌液的总最终浓度,在混合该菌株混合物及该金刺梨样品后为1.5×107CFU/ml~3.5×107CFU/ml,且混合该菌株混合物及该金刺梨样品后,该菌株混合物能够占总体积的10~25%,借此能够提供足够的菌数来进行发酵,确保具有预期的发酵效率。
将该菌株混合物于进行发酵反应的最适条件下发酵该金刺梨样品,借此以获得该金刺梨发酵液。详言之,将该菌株混合物在22~33℃的温度下发酵该金刺梨样品6~15天,且发酵的过程中能够采静置发酵,亦能够以80~110 rpm(转)的震荡培养来进行发酵,以防止沉淀产生。本实施例中,将该菌株混合物于28℃下发酵该金刺梨样品14天,并以100 rpm的震荡培养以提升发酵效果。
为证明本发明的金刺梨发酵方法确实能够获得提升抗氧化能力的该金刺梨发酵液,现进行以下实验。
以0.8:0.8:1.6的菌数比,混合该干酪乳酸杆菌菌株的菌液、该长双歧杆菌菌株的菌液及该酿酒酵母菌菌株的菌液,以获得该菌株混合物,以该菌株混合物于28℃下发酵该金刺梨样品14天后以形成该金刺梨发酵液,将该金刺梨发酵液进行离心以将杂质分离后,吸取一上清液后作为实验组(A1)。另外将与该干酪乳酸杆菌同属但不同种的一嗜酸乳杆菌(Lactobacillus acidophilus)菌株的菌液、加上该长双歧杆菌菌株的菌液及该酿酒酵母菌菌株的菌液,以0.8:0.8:1.6的菌数比混合并共同发酵该金刺梨样品后,以同样的方法处理做为第一对照组(A2),以及未经发酵的该金刺梨样品做为第二对照组(A3),分别检测各组(A1~A3)的抗氧化能力及抗氧化物质含量。
(A)抗氧化能力测定
ABTS(2,2'-Azino-bis -〔3-ethylbenthiazoline sulfonic acid〕)为一种显色剂,当ABTS与氧化剂及过氧化氢(H2O2)反应后会产生ABTS+,ABTS+为一种稳定的蓝绿色物质,并于734nm有吸收光,当抗氧化剂加入后会抑制ABTS+的产生,因此该ABTS+的吸光值越低,表示该抗氧化剂的抗氧化能力越佳。因此,将上述实验组(A1)与第一对照组(A2)及第二对照组(A3)稀释500倍后,比较各组对于ABTS+的清除率,结果如图1a所示,使用包括该干酪乳酸杆菌菌株、该长双歧杆菌菌株及该酿酒酵母菌菌株的菌株混合物发酵的该金刺梨发酵液(A1),相较于以该嗜酸乳杆菌菌株、该长双歧杆菌菌株及该酿酒酵母菌菌株共同发酵的该金刺梨发酵液(A2),及未经发酵的该金刺梨样品(A3),ABTS+的清除率均有显著的提升。
另外,将上述第A1~A3组稀释100倍后,测定其中所含的超氧化物歧化酶(SOD)的活性,结果如图1b所示,以包括该干酪乳酸杆菌菌株、该长双歧杆菌菌株及该酿酒酵母菌菌株的菌株混合物发酵的该金刺梨发酵液(A1),其超氧化物歧化酶活性显著高于第A2、A3组。
(B)抗氧化物质含量测定
继续将上述第A1~A3组稀释500倍后,测定其中所含有的维生素C(Vitamin C)及类胡萝卜素(Carotenoid)的含量,结果如图2a、2b所示,试验结果显示,第A1组的维生素C及类胡萝卜素含量,明显高于第A2、A3组。因此能够证实,经该干酪乳酸杆菌菌株、该长双歧杆菌菌株及该酿酒酵母菌菌株共同发酵所获得的金刺梨发酵液,其中所含有的维生素C及类胡萝卜素的含量确实有增加。
因此,经该干酪乳酸杆菌菌株、该长双歧杆菌菌株及该酿酒酵母菌菌株共同发酵该金刺梨样品后的金刺梨发酵液,其抗氧化能力及抗氧化物质含量,相较于未经发酵的金刺梨样品确实具有显著提升,且亦高于该嗜酸乳杆菌菌株、该长双歧杆菌菌株及该酿酒酵母菌菌株共同发酵该金刺梨样品后的金刺梨发酵液,因此能够证实,本发明使用该干酪乳酸杆菌菌株、该长双歧杆菌菌株及该酿酒酵母菌菌株共同发酵该金刺梨样品,能够获得提升抗氧化能力的该金刺梨发酵液。
(C)发酵菌株浓度的影响
本实验在以0.8:0.8:1.6的菌数比,混合该干酪乳酸杆菌菌株的菌液、该长双歧杆菌菌株的菌液及该酿酒酵母菌菌株的菌液,以形成该菌株混合物,并混合该菌株混合物及该金刺梨样品后,使该干酪乳酸杆菌菌株的菌液、该长双歧杆菌菌株的菌液及该酿酒酵母菌菌株的菌液的总最终浓度分别形成为3×107CFU/ml及1×107CFU/ml,并于28℃下分别发酵该金刺梨样品14天以形成两组金刺梨发酵液(B1及B2),并以未经发酵的该金刺梨样品(B3)作为对照组,分别检测各组(B1~B3)的抗氧化能力及抗氧化物质含量,以比较不同浓度的菌株对发酵该金刺梨样品的影响;其结果如图3a-3d所示,第B1组的ABTS+的清除率及超氧化物歧化酶(SOD)的活性皆有显著的提升,且第B1组的维生素C及类胡萝卜素含量亦增加,显示在混合该菌株混合物及该金刺梨样品后,以总最终浓度3×107CFU/ml的该干酪乳酸杆菌菌株的菌液、该长双歧杆菌菌株的菌液及该酿酒酵母菌菌株的菌液发酵该金刺梨样品,能够有效的提升该金刺梨样品的抗氧化能力。
(D)发酵温度的影响
本实验在以0.8:0.8:1.6的菌数比,混合该干酪乳酸杆菌菌株的菌液、该长双歧杆菌菌株的菌液及该酿酒酵母菌菌株的菌液,以形成该菌株混合物,并混合该菌株混合物及该金刺梨样品后,使该干酪乳酸杆菌菌株的菌液、该长双歧杆菌菌株的菌液及该酿酒酵母菌菌株的菌液的总最终浓度形成3×107CFU/ml,并分别于28℃及20℃下,发酵该金刺梨样品14天以形成两组金刺梨发酵液(C1及C2),并以未经发酵的该金刺梨样品(C3)作为对照组,分别检测各组(C1~C3)的抗氧化能力及抗氧化物质含量,以比较在不同温度对发酵该金刺梨样品的影响;其结果如图4a-4d所示,第C1组的ABTS+的清除率及超氧化物歧化酶(SOD)的活性皆有显著的提升,且第C1组的维生素C及类胡萝卜素含量亦增加,显示于28℃下发酵相较于20℃下发酵,能够获得高抗氧化能力的该金刺梨发酵液。
(E)发酵时间的影响
本实验在以0.8:0.8:1.6的菌数比,混合该干酪乳酸杆菌菌株的菌液、该长双歧杆菌菌株的菌液及该酿酒酵母菌菌株的菌液,以形成该菌株混合物,并混合该菌株混合物及该金刺梨样品后,使该干酪乳酸杆菌菌株的菌液、该长双歧杆菌菌株的菌液及该酿酒酵母菌菌株的菌液的总最终浓度形成3×107CFU/ml,并于28℃下分别发酵该金刺梨样品14及5天,以形成两组金刺梨发酵液(D1及D2),并以未经发酵的该金刺梨样品(D3)作为对照组,分别检测各组(D1~D3)的抗氧化能力及抗氧化物质含量,以比较在不同发酵时间对发酵该金刺梨样品的影响;其结果如图5a-5d所示,第D1组的ABTS+的清除率及超氧化物歧化酶(SOD)的活性,相较于第D2组及第D3组皆有显著的提升,且第D1组的维生素C及类胡萝卜素含量,相较于第D2组及第D3组亦增加,并且,仅发酵该金刺梨样品5天(D2)所获得的金刺梨发酵液的抗氧化能力与抗氧化物质含量,与对照组(D3)的抗氧化能力与抗氧化物质含量相比皆无明显的变化,进一步显示本发明的金刺梨发酵方法发酵该金刺梨样品6~15天,可以有效的提升该金刺梨样品的抗氧化能力。
综上所述,本发明的金刺梨发酵方法,通过以包括该干酪乳酸杆菌菌株、该长双歧杆菌菌株及该酿酒酵母菌菌株的菌株混合物,发酵该金刺梨样品,使该金刺梨样品的抗氧化活性及所含的抗氧化物质含量得以提升,以获得提升抗氧化能力的该金刺梨发酵液,能够作为抗氧化保健食品食用或作为食品添加物使用,达到食用者养生的功效。
Claims (1)
1.一种金刺梨发酵方法,其特征在于,包括:
提供一个菌株混合物,该菌株混合物系分别将该干酪乳酸杆菌菌株BCRC10697、该长双歧杆菌菌株BCRC14604及该酿酒酵母菌菌株BCRC20579置于一液态培养基中进行增殖培养,并于37℃中培养至对数生长期后分别形成一干酪乳酸杆菌菌株的菌液,一长双歧杆菌菌株的菌液及一酿酒酵母菌菌株的菌液,并以0.8:0.8:1.6的菌数比混合该干酪乳酸杆菌菌株的菌液、该长双歧杆菌菌株的菌液及该酿酒酵母菌菌株的菌液;
提供一个金刺梨样品,将该金刺梨样品进行榨汁形成一金刺梨汁;及
以该菌株混合物发酵该金刺梨汁,使该干酪乳酸杆菌菌株的菌液、该长双歧杆菌菌株的菌液及该酿酒酵母菌菌株的菌液的总最终浓度在混合该菌株混合物及该金刺梨汁后为1.5×107CFU/ml~3.5×107CFU/ml,且混合该菌株混合物及该金刺梨汁后,该菌株混合物占总体积的10~25%,于28℃的温度并以100rpm的转速震荡培养发酵该金刺梨汁14天,以获得一个金刺梨发酵液。
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