CN115624181A - 组合物、制造方法及应用 - Google Patents
组合物、制造方法及应用 Download PDFInfo
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- CN115624181A CN115624181A CN202211398838.1A CN202211398838A CN115624181A CN 115624181 A CN115624181 A CN 115624181A CN 202211398838 A CN202211398838 A CN 202211398838A CN 115624181 A CN115624181 A CN 115624181A
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- powder
- oil
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- bacteria
- bifidobacterium
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Abstract
提供:包含菌粉和脂肪油且前述菌粉的结块得到抑制的组合物、制造方法及应用;包含菌粉、脂肪油、以及选自由微粉和表面活性剂组成的组中的至少1种的组合物;其制造方法以及选自由微粉和表面活性剂组成的组中的至少1种作为菌粉用抗结块剂在包含脂肪油的组合物中的应用。
Description
本申请是申请日为2020年3月19日、申请号为202080057079.5,发明名称为“组合物、制造方法及应用”的申请的分案申请。
技术领域
本发明涉及组合物、制造方法及应用。
本申请主张基于2019年10月17日在美国申请的临时专利申请第62916346号的优先权,将其内容援用至此。
背景技术
报道了:菌具有便秘和腹泻的改善效果、乳糖不耐症的改善效果、基于免疫功能改善的感染防御和变态反应抑制效果、动脉硬化的预防效果、以及抗肿瘤作用等对于人类的健康有益的效果。因此,近些年,市场上有将菌粉悬浮于脂肪油中而成的被称为油滴(oildrop)的产品。
油滴中,期望的是将菌粉均匀地分散在脂肪油中,但有时菌粉会沉淀和固化在容器的底部,最终形成菌粉难以再分散的沉积体(结块)。
专利文献1中记载了包含益生菌细菌的1种以上或1株以上、油、以及无水磷酸氢钙的补充剂组合物。记载了:在该补充剂组合物中,维持了益生菌细菌的存活率。该补充剂组合物是以油作为分散介质的悬浮液。
然而,并不清楚专利文献1中记载的补充剂组合物中是否抑制了益生菌细菌的结块。
现有技术文献
专利文献
专利文献1:美国专利申请公开第2018/0235271号说明书
发明内容
发明要解决的问题
根据本发明人等的研究,如后述的参考例所示,仅将菌粉中包含的载体分散在作为分散介质的脂肪油中的情况,未发生前述载体的结块。本发明人等由该研究结果考虑出油滴中的菌粉的结块是由菌粉中包含的菌体本身所引起的。
本发明的目的在于提供包含菌粉和脂肪油、且前述菌粉的结块得到抑制的组合物、制造方法及应用。
用于解决问题的方案
本发明人等为了解决前述课题而进行了深入研究的结果,得到通过以下的构成而能够解决前述课题,完成了本发明。
[1]一种组合物,其包含:菌粉、脂肪油、以及选自由微粉和表面活性剂组成的组中的至少1种。
[2]根据[1]所述的组合物,其为悬浮液。
[3]根据[1]或[2]所述的组合物,其中,前述菌粉包含选自由乳酸菌的活菌、乳酸菌的死菌、双歧杆菌属细菌的活菌和双歧杆菌属细菌的死菌组成的组中的至少1种。
[4]根据[1]~[3]中任一项所述的组合物,其中,前述微粉包含选自由微晶纤维素和磷酸三钙组成的组中的至少1种。
[5]根据[1]~[4]中任一项所述的组合物,其中,前述表面活性剂包含选自由阴离子表面活性剂和HLB值低于7的非离子表面活性剂组成的组中的至少1种。
[6]根据[5]所述的组合物,其中,前述阴离子表面活性剂包含选自由硬脂酸金属盐组成的组中的至少1种,前述HLB值低于7的非离子表面活性剂包含选自由HLB值低于7的蔗糖脂肪酸酯和HLB值低于7的甘油脂肪酸酯组成的组中的至少1种。
[7]根据[1]~[6]中任一项所述的组合物,其包含相对于前述组合物的总质量为0.5~10质量%的前述菌粉。
[8]根据[1]~[7]中任一项所述的组合物,其还包含添加剂。
[9]一种[1]~[8]中任一项所述的组合物的制造方法,其中,按任意的顺序将菌粉、脂肪油、以及选自由微粉和表面活性剂组成的组中的至少1种进行混合;或者,在包含菌粉和脂肪油的悬浮液中混合选自由微粉和表面活性剂组成的组中的至少1种。
[10]选自由微粉和表面活性剂组成的组中的至少1种作为菌粉用抗结块剂在包含脂肪油的组合物中的应用。
发明的效果
根据本发明,可以提供包含菌粉和脂肪油、且前述菌粉的结块得到抑制的组合物、制造方法及应用。
对于本发明的组合物,菌粉的结块得到抑制、菌粉容易分散在作为分散介质的脂肪油中。
具体实施方式
结块是指:在包含菌粉的悬浮液中,菌粉沉淀和固化并形成难以再分散的沉积体。即,结块是指发生沉淀和固化且形成难以再分散的沉积体,若即使沉淀和固化也能再分散,则不能称为结块。
使用“~”所表示的数值范围包括该数值范围的上限值和下限值。
[组合物]
本发明的组合物包含菌粉、脂肪油、以及选自由微粉和表面活性剂组成的组中的至少1种。
以下对菌粉、脂肪油、微粉和表面活性剂详细地进行说明。
〈菌粉〉
菌粉(Cell powder)是干燥的菌体的统称。
作为将菌体干燥的方法,例如可以使用冷冻干燥或喷雾干燥,但不限定于这些。
冷冻干燥是使用冷冻干燥装置、液氮等在约-20℃~-160℃左右的低温下通常以1~60Pa左右的减压下进行干燥的方法。
喷雾干燥是利用微粒化装置使液体变为液滴,将该液滴喷雾在相对高温的加热气流中,由此使水分蒸发进行干燥的方法。
菌粉可以是仅包含菌体的状态、或是包含菌体及除菌体以外的成分的状态。作为除前述菌体以外的成分,例如可以使用冷冻保护剂、冷冻干燥保护剂、喷雾干燥保护剂和载体。
菌粉可以是分散在作为药物的原材料或饮食品的原材料具有使用效果的研制剂(日文:倍散剤)、赋形剂或载体中的状态。作为前述研制剂、前述赋形剂或前述载体,例如可列举出淀粉、淀粉分解物或糊精。作为前述淀粉,例如可列举出玉米淀粉、马铃薯淀粉或木薯淀粉。
菌粉优选包含选自由乳酸菌的活菌、乳酸菌的死菌、双歧杆菌属细菌的活菌和双歧杆菌属细菌的死菌组成的组中的至少1种。
《乳酸菌》
乳酸菌是归属于细菌领域的厚壁菌门且通过代谢而产生乳酸的细菌的统称。
作为乳酸菌,在归属于厚壁菌门且通过代谢而产生乳酸的细菌中,优选归属于芽孢杆菌纲乳杆菌目的乳酸菌,更优选归属于气球菌科(Aerococcaceae)、肉杆菌科(Carnobacterium)、肠球菌科(Enterococcus)、链球菌科(Streptococcus)、乳杆菌科(Lactobacillus)、明串珠菌科(Leuconostocaceae)或链球菌科(Streptococcaceae)的乳酸菌。
作为归属于乳杆菌科的乳酸菌,可列举出加氏乳杆菌(Lactobacillus gasseri)、嗜酸乳杆菌(L.acidophilus)、瑞士乳杆菌(L.helveticus)、副干酪乳杆菌(L.paracasei)、干酪乳杆菌(L.casei)、鼠李糖乳杆菌(L.rhamunosus)、德氏乳杆菌(L.delbrueckii)、德氏乳杆菌保加利亚亚种(L.derubruekii subsp.bulgaricus)和植物乳杆菌(L.plantarum)等乳杆菌属细菌。
作为归属于肠球菌科的乳酸菌,可列举出粪肠球菌(Enterococcus faecalis)和屎肠球菌(E.faecium)等肠球菌属细菌。
作为归属于链球菌科(Streptococci)的乳酸菌,可列举出乳酸乳球菌(Lactococcus lactis)、乳酸乳球菌乳脂亚种(L.lactis subsp.cremoris)等乳球菌属细菌、嗜热链球菌(Streptococcus thermophilus)等链球菌属细菌。
作为归属于明串珠菌科的乳酸菌,可列举出肠膜明串珠菌(Leuconostocmesenteroides)、肠膜明串珠菌乳脂亚种(L.mesenteroides subsp.cremoris)等明串珠菌属细菌。
作为乳酸菌,优选选自由上述菌种组成的组中的至少1种。另外,作为乳酸菌,也可以使用属名(genus)或种加词(epithet)未确定的菌株、或新种菌株。
乳酸菌可以在活菌和死菌的任意状态下使用。乳酸菌的形态可以使用经冷冻者、经冷冻干燥者或经喷雾干燥者。进而,乳酸菌也可以是仅包含乳酸菌的菌体、乳酸菌的菌体中包含除菌体以外的成分(例如,冷冻保护剂、冷冻干燥保护剂、喷雾干燥保护剂等)的状态。另外,也可以是分散在研制剂中的状态。作为研制剂,可以使用玉米淀粉、马铃薯淀粉、木薯淀粉等淀粉、淀粉分解物、糊精、麦芽糊精等。
(加氏乳杆菌)
加氏乳杆菌只要对宿主发挥有益的效果,就没有特别限制。具体而言,加氏乳杆菌通过单独使用、或与其它有效成分的组合使用,只要对宿主发挥有益的效果,就没有特别限制。
作为加氏乳杆菌,具体可列举出NITE BP-01669、ATCC 33323、DSM 20243、JCM1131、SBT 2055和OLL 2716。作为加氏乳杆菌,特别优选NITE BP-01669。加氏乳杆菌可以单独使用1株,或者也可以组合2株以上而使用。
(嗜酸乳杆菌)
嗜酸乳杆菌只要对宿主发挥有益的效果,就没有特别限制。具体而言,嗜酸乳杆菌通过单独使用、或与其它有效成分的组合使用,只要对宿主发挥有益的效果,就没有特别限制。
作为嗜酸乳杆菌,具体可列举出NITE BP-01695、ATCC 4356、DSM 20079、JCM1132、YIT 0168和YIT 0154。作为嗜酸乳杆菌,特别优选NITE BP-01695。嗜酸乳杆菌可以单独使用1株,或者也可以组合2株以上而使用。
(瑞士乳杆菌)
瑞士乳杆菌只要对宿主发挥有益的效果,就没有特别限制。具体而言,瑞士乳杆菌通过单独使用、或与其它有效成分的组合使用,只要对宿主发挥有益的效果,就没有特别限制。
作为瑞士乳杆菌,具体可列举出NITE BP-01671、ATCC 15009、DSM 20075、JCM1120和SBT 2171。作为瑞士乳杆菌,特别优选NITE BP-01671。瑞士乳杆菌可以单独使用1株,或者也可以组合2株以上而使用。
(副干酪乳杆菌)
副干酪乳杆菌只要对宿主发挥有益的效果,就没有特别限制。具体而言,副干酪乳杆菌通过单独使用、或与其它有效成分的组合使用,只要对宿主发挥有益的效果,就没有特别限制。
作为副干酪乳杆菌,具体可列举出NITE BP-01633、ATCC 25302、DSM 5622、JCM8130、ATCC 25599、DSM 20258和JCM 1171。作为副干酪乳杆菌,特别优选NITE BP-01633。副干酪乳杆菌可以单独使用1株,或者也可以组合2株以上而使用。
(乳酸菌的培养方法)
乳酸菌的菌体可以通过培养乳酸菌而容易地获得。培养方法只要能使乳酸菌增殖就没有特别限定。作为培养方法,例如,可以直接使用通常用于乳酸菌的培养的方法、或进行适宜修正而使用。培养温度优选25~50℃,更优选35~42℃。培养优选在厌氧条件下进行。例如,可以边通入二氧化碳气等非氧化性气体边进行培养。另外,培养也可以在液体静置培养等微好氧条件下进行。培养例如进行至乳酸菌增殖到期望的程度即可。
培养中使用的培养基只要能使乳酸菌增殖就没有特别限定。作为培养基,例如可以直接使用通常用于乳酸菌的培养的培养基,或者也可以进行适宜修正而使用。即,作为碳源,例如可以根据同化性使用半乳糖、葡萄糖、果糖、甘露糖、纤维素二糖、麦芽糖、乳糖、蔗糖、海藻糖、淀粉、淀粉水解物、废糖蜜等糖类。另外,也可以在包含酪蛋白、乳清、或其分解物等乳蛋白的培养基中进行培养。作为氮源,例如可以使用氨、硫酸铵、氯化铵、硝酸铵等铵盐或硝酸盐。另外,作为无机盐类,例如可以使用氯化钠、氯化钾、磷酸钾、硫酸镁、氯化钙、硝酸钙、氯化锰、硫酸亚铁等。另外,可以使用蛋白胨、大豆粉、脱脂大豆粕、肉提取物、酵母提取物等有机成分。另外,作为已制备完成的培养基,例如可以使用MRS培养基(de Man,Rogosa,and Sharpe medium)。
《双歧杆菌属细菌》
双歧杆菌(Bifidobacterium)属是属于细菌领域的放线菌门放线菌纲双歧杆菌目的一组细菌的属名。
作为双歧杆菌属细菌,例如可列举出长双歧杆菌婴儿亚种(B.longumsubsp.infantis)、短双歧杆菌(B.breve)、长双歧杆菌长亚种(B.longum subsp.longum)、长双歧杆菌猪亚种(B.longum subsp.suis)、动物双歧杆菌乳亚种(B.animalissubsp.lactis)、动物双歧杆菌动物亚种(B.animalis subsp.animalis)、两歧双歧杆菌(B.bifidum)、青春双岐杆菌(B.adolescentis)、角双歧杆菌(B.angulatum)、齿双歧杆菌(B.dentium)、假小链双歧杆菌(B.pseudocatenulatum)、伪长双歧杆菌(B.pseudolongum)和嗜热双歧杆菌(B.thermophilum)。
作为双歧杆菌属细菌,优选使用选自由长双歧杆菌婴儿亚种(B.longumsubsp.infantis)、短双歧杆菌(B.breve)、长双歧杆菌长亚种(B.longum subsp.longum)、长双歧杆菌猪亚种(B.longum subsp.suis)、动物双歧杆菌乳亚种(B.animalissubsp.lactis)、和两歧双歧杆菌(B.bifidum)组成的组中的至少1种。更优选使用选自由长双歧杆菌婴儿亚种(B.longum subsp.infantis)、短双歧杆菌(B.breve)、长双歧杆菌长亚种(B.longum subsp.longum)、动物双歧杆菌乳亚种(B.animalis subsp.lactis)组成的组中的至少1种。
双歧杆菌属细菌可以单独使用1种或组合两种以上而使用。
作为组合使用双歧杆菌属细菌时的组合,优选长双歧杆菌的各亚种中的1种以上与短双歧杆菌的组合。
作为双歧杆菌属细菌,也可以使用种加词(epithet)未确定的菌株、或新种菌株。
双歧杆菌属细菌可以在活菌和死菌的任意状态下使用。双歧杆菌属细菌的形态可以使用经冷冻者、经冷冻干燥者或经喷雾干燥者。进而,双歧杆菌属细菌也可以是仅包含双歧杆菌属细菌的菌体的状态、双歧杆菌属细菌的菌体中包含除菌体以外的成分(例如,冷冻保护剂、冷冻干燥保护剂、喷雾干燥保护剂等)的状态。另外,也可以是分散在研制剂中的状态。作为研制剂,可以使用玉米淀粉、马铃薯淀粉、木薯淀粉等淀粉、淀粉分解物、糊精、麦芽糊精等。
(长双歧杆菌婴儿亚种)
长双歧杆菌婴儿亚种只要对宿主发挥有益的效果,就没有特别限制。具体而言,长双歧杆菌婴儿亚种通过单独使用、或与其它有效成分的组合使用,只要对宿主发挥有益的效果,就没有特别限制。
作为长双歧杆菌婴儿亚种,具体可列举出NITE BP-02623、ATCC 15697、ATCC15702、DSM 20088和JCM 1222。作为长双歧杆菌婴儿亚种,特别优选NITE BP-02623。长双歧杆菌婴儿亚种可以单独使用1株,或者也可以组合2株以上而使用。
(短双歧杆菌)
短双歧杆菌只要对宿主发挥有益的效果,就没有特别限制。具体而言,短双歧杆菌通过单独使用、或与其它有效成分的组合使用,只要对宿主发挥有益的效果,就没有特别限制。
作为短双歧杆菌,具体例如可列举出NITE BP-02622、FERM BP-11175、ATCC15700、ATCC 15698、DSM 20213、DSM 24706、DSM 13692、DSM 24732、DSM 24736、DSM16604、JCM 1192、NCC 2705、NCC 490、YIT 4010、YIT 4064、SBT 2928、UCC 2003、BBG-001、C50、R0070、BG7。作为短双歧杆菌,特别优选NITE BP-02622。短双歧杆菌可以单独使用1株,或者也可以组合2株以上而使用。
(长双歧杆菌长亚种)
长双歧杆菌长亚种只要对宿主发挥有益的效果,就没有特别限制。具体而言,长双歧杆菌长亚种通过单独使用、或与其它有效成分的组合使用,只要对宿主发挥有益的效果,就没有特别限制。
作为长双歧杆菌长亚种,具体例如可列举出NITE BP-02621、ATCC 15707、ATCC25962、DSM 20219和JCM 1217。作为长双歧杆菌长亚种,特别优选NITE BP-02621。长双歧杆菌长亚种可以单独使用1株,或者也可以组合2株以上而使用。
(长双歧杆菌猪亚种)
长双歧杆菌猪亚种只要对宿主发挥有益的效果,就没有特别限制。具体而言,长双歧杆菌猪亚种通过单独使用、或与其它有效成分的组合使用,只要对宿主发挥有益的效果,就没有特别限制。
作为长双歧杆菌猪亚种,具体例如可列举出ATCC 27533、ATCC 27532、DSM 20211和JCM 1269。作为长双歧杆菌猪亚种,特别优选ATCC 27533。长双歧杆菌猪亚种可以单独使用1株,或者也可以组合2株以上而使用。
(动物双歧杆菌乳亚种)
动物双歧杆菌乳亚种只要对宿主发挥有益的效果,就没有特别限制。具体而言,动物双歧杆菌乳亚种通过单独使用、或与其它有效成分的组合使用,只要对宿主发挥有益的效果,就没有特别限制。
作为动物双歧杆菌乳亚种,具体例如可列举出DSM 15954和FERM P-21998。作为动物双歧杆菌乳亚种,特别优选DSM 15954。动物双歧杆菌乳亚种可以单独使用1株,或者也可以组合2株以上而使用。
需要说明的是,代替由示例的菌株编号确定的菌株,也可以使用与以该菌株编号保存在菌株保存机构(典型培养物保藏中心)中的菌株处于实质上相同菌株关系者。例如,若为长双歧杆菌长亚种,则可以代替ATCC 15707使用DSM 20219或JCM 1217。作为是否处于实质上相同菌株关系的指标,例如,可以使用16S rRNA基因碱基序列的同一性。菌株处于实质上相同关系时的16S rRNA基因碱基序列的同一性优选99.86%以上,更优选99.93%以上,进一步优选100%。菌株处于实质上相同关系时,特别优选16S rRNA基因碱基序列的同一性为100%、且同化性等微生物学性质相同。
另外,代替由示例的菌株编号确定的菌株,也可以使用该菌株的衍生株。作为衍生株,可列举出从保存株经人工培育的菌株、从保存株自然生长出的菌株。作为培育的方法,可列举出基于基因工程方法的改变、基于突变处理的改变。作为突变处理,例如可列举出X射线的照射、紫外线的照射、或基于N-甲基-N’-硝基-N-亚硝基胍(MNNG)、乙基甲磺酸盐(EMS)或甲基甲磺酸盐(MMS)等诱变剂的处理。作为从保存株自然生长出的菌株,可列举出在使用保存株时自然生长出的突变株。衍生株可以通过1株的改变来构建,也可以通过2株以上的改变来构建。
作为双歧杆菌属细菌,可以使用市售品,也可以使用适宜制造而获得者。作为市售品,例如可列举出长双歧杆菌长亚种NITE BP-02621、短双歧杆菌NITE BP-02622、长双歧杆菌婴儿亚种NITE BP-02623、动物双歧杆菌乳亚种BB-12(DSM 15954)。
(双歧杆菌属细菌的培养方法)
双歧杆菌属细菌的菌体可以通过培养双歧杆菌属细菌而容易地获得。培养方法只要能使双歧杆菌属细菌增殖就没有特别限定。作为培养方法,例如可以直接使用通常用于双歧杆菌属细菌的培养的方法、或进行适宜修正而使用。培养温度优选25~50℃,更优选35~42℃。培养优选在厌氧条件下进行。例如,可以边通入二氧化碳气等非氧化性气体边进行培养。另外,培养也可以在液体静置培养等微好氧条件下进行。培养例如进行至双歧杆菌属细菌增殖到期望的程度即可。
培养中使用的培养基只要能使双歧杆菌属细菌增殖就没有特别限定。作为培养基,例如可以直接使用通常用于双歧杆菌属细菌的培养的培养基,或者也可以进行适宜修正而使用。即,作为碳源,例如可以根据同化性使用半乳糖、葡萄糖、果糖、甘露糖、纤维素二糖、麦芽糖、乳糖、蔗糖、海藻糖、淀粉、淀粉水解物、废糖蜜等糖类。另外,也可以在包含酪蛋白、乳清、或其分解物等乳蛋白的培养基中进行培养。作为氮源,例如可以使用氨、硫酸铵、氯化铵、硝酸铵等铵盐或硝酸盐。另外,作为无机盐类,例如可以使用氯化钠、氯化钾、磷酸钾、硫酸镁、氯化钙、硝酸钙、氯化锰、硫酸亚铁等。另外,可以使用蛋白胨、大豆粉、脱脂大豆粕、肉提取物、酵母提取物等有机成分。另外,作为已制备完成的培养基,例如可以使用MRS培养基(de Man,Rogosa,and Sharpe medium)。
《菌种保藏的首字母缩略词》
菌种保藏的首字母缩略词如下所述。
NITE:国家技术评估学会,专利微生物保藏中心(NITE Patent MicroorganismsDepositary,NPMD),日本国家标准与技术研究院(National Institute of Technologyand Evaluation)
FERM:国家技术评估学会,专利微生物保藏中心(NITE Patent MicroorganismsDepositary,NPMD),日本国家标准与技术研究院(National Institute of Technologyand Evaluation)
ATCC:美国菌种保藏中心(American Type Culture Collection)
DSM:德国微生物菌种保藏中心(Deutsche Sammlung von Mikroorganismen undZellkulturen GmbH,DSMZ)
JCM:JCM菌株保藏中心(Japan Collection of Microorganisms),Riken BRC
NCC:雀巢公司(Nestle S.A.)
YIT:养乐多本社株式会社(Yakult Honsha Co.,Ltd.)
SBT:雪印惠乳业株式会社(MEGMILK Snow Brand Co.,Ltd.)
OLL:株式会社明治(Meiji Co.,Ltd.)
R:拉曼集团(LALLEMAND Inc.)
《益生菌》
菌粉可以是活菌或死菌,但菌粉为活菌时,可以期待发挥作为“益生菌”的功能。
需要说明的是,益生菌是相对于抗生素(antibiotics)而提出的术语,词源是具有共生含义的源于共生(probiosis)。目前,被广泛接受的益生菌的定义是“通过改善肠道菌群的平衡而对宿主的健康产生积极影响的活微生物”。
《菌粉的含量》
本发明的组合物中的菌粉的含量相对于组合物的总质量优选0.5~10质量%,更优选1~5质量%。
菌粉的含量相对于组合物的总质量为0.5质量%以上时,能够进一步发挥本发明的组合物含有菌粉所带来的有益效果。
菌粉的含量相对于组合物的总质量为10质量%以下时,使本发明的组合物含有菌粉所带来的有益效果与成本的平衡变得更良好。
〈脂肪油〉
本发明中的脂肪油在本发明的组合物流通的过程中例如优选在0~40℃的至少一部分范围内为液体的油,更优选在0~40℃的整个范围内为液体的油。
作为脂肪油,优选食用油。
作为食用油,例如可列举出榛子油、橄榄油、月见草油、南瓜油、米糠油、大豆油、玉米油、向日葵籽油、菜籽油、红花油(Safflower oil)、椰油(包括羽叶棕榈油、锯棕榈油等)、棕榈油、棕榈仁油、中链脂肪酸甘油三酯(MCT)、二十二碳六烯酸(DHA)、二十碳五稀酸(EPA)、亚麻仁油、紫苏籽油(紫苏油)、米胚芽油、小麦胚芽油、椰子油、棉籽油、花生油、芝麻油、杏仁油、腰果油、澳洲胡桃油、蒙刚果油、山核桃油、松子油、阿月浑子树油、胡桃油、葫芦籽油、旱地油瓜籽油、南瓜籽油、西瓜籽油、巴西莓提取物、黑醋栗油、琉璃苣油、月见草油、苋菜油、杏仁油、苹果油、摩洛哥坚果油、洋蓟油、鳄梨油、巴巴苏仁油、辣木油、板栗油(Yangu oil)、角豆树油、芫荽油、麦芽油(Dica oil)、亚麻油、葡萄籽油、大麻油、木棉籽油、扁柄草油(Lallemantia oil)、马鲁拉油、白芒花油、芥子油、秋葵油(木槿油)、木瓜油、罂粟油、欧洲李油、藜麦油、皂厂杂油、山茶籽油(Camellia oil)、蓟油、番茄油、磷虾油(Krilloil)和琉璃苣油。
作为脂肪油,优选选自由橄榄油、米糠油、大豆油、玉米油、向日葵籽油、红花油(Safflower oil)、中链脂肪酸甘油三酯(MCT)组成的组中的至少1种,更优选选自由玉米油、向日葵籽油、中链脂肪酸甘油三酯(MCT)组成的组中的至少1种,特别优选中链脂肪酸甘油三酯(MCT)。
脂肪油相对于组合物的总质量优选包含90质量%以上,更优选包含92质量%以上,进一步优选包含95质量%以上。
〈微粉和表面活性剂〉
本发明的组合物中,微粉和表面活性剂作为抗结块剂发挥作用。
抗结块剂具有防止或消除结块的作用。
《微粉》
微粉是有机物或无机物的微小尺寸的粉末,优选有机物的微小尺寸的粉末。
作为有机物的微小尺寸的粉末,例如可列举出微晶纤维素。微晶纤维素是用酸将浆粕水解和纯化而得到的高纯度的纤维素。
微晶纤维素颗粒的平均粒径以D50计优选1~200μm,更优选5~100μm。
微小颗粒的平均粒径(D50)是由利用激光衍射/散射法求出的体积分布计算出的50%体积粒径。
作为无机物的微小尺寸的粉末,例如可列举出磷酸三钙、和微粒二氧化硅。磷酸三钙是由化学式Ca3(PO4)2所示的磷酸与钙的盐。磷酸三钙具有三种多晶型,但在本发明的组合物中,观察到作为低温多晶型的β-磷酸三钙(β-TCP)。微粒二氧化硅是二氧化硅的微小颗粒。作为无机物的微小尺寸的粉末,特别优选磷酸三钙。
磷酸三钙颗粒的平均粒径以D50计优选1~200μm,更优选1~50μm。
微粉优选包含选自由微晶纤维素和磷酸三钙组成的组中的至少1种。
本发明的组合物中,微粉和表面活性剂适于用作菌粉用抗结块剂。
《表面活性剂》
表面活性剂优选选自由阴离子表面活性剂和HLB值低于7的非离子表面活性剂组成的组中的至少1种。
(阴离子表面活性剂)
作为阴离子表面活性剂,例如可列举出羧酸型阴离子表面活性剂、直链烷基苯磺酸钠、磺酸型阴离子表面活性剂、硫酸酯型阴离子表面活性剂和磷酸酯型阴离子表面活性剂。
作为阴离子表面活性剂,优选选自由碳数为12~18的脂肪酸的盐组成的组中的至少1种,更优选选自由硬脂酸金属盐组成的组中的至少1种,进一步优选硬脂酸的碱土金属盐,特别优选硬脂酸钙或硬脂酸镁。
阴离子表面活性剂可以使用单独1种,或者组合两种以上而使用。
(HLB值低于7的非离子表面活性剂)
作为HLB值低于7的非离子表面活性剂,优选HLB值低于7的脂肪酸酯,更优选选自由HLB值低于7的蔗糖脂肪酸酯和HLB值低于7的甘油脂肪酸酯组成的组中的至少1种,进一步优选选自由HLB值低于7的蔗糖脂肪酸酯组成的组中的至少1种。
HLB值低于7的非离子表面活性剂可以使用单独1种,或者组合两种以上而使用。
《微粉和表面活性剂的含量》
本发明的组合物中的微粉和表面活性剂的含量以微粉和表面活性剂的总计计、相对于组合物的总质量优选0.01~10质量%,更优选0.1~5质量%,进一步优选0.5~2质量%。
〈添加剂〉
本发明的组合物除了上述成分之外可以进一步包含添加剂。
作为添加剂,可列举出抗氧化剂、赋形剂、结合剂、崩解剂、润滑剂、稳定剂、矫味矫臭剂、稀释剂等。
作为抗氧化剂,优选维生素E。维生素E是脂溶性维生素,容易溶解于组合物中的脂肪油中。
〈组合物的用途〉
本发明的组合物例如可以是补充剂组合物、饮料组合物、食品组合物、药物组合物、饲料组合物,但优选用作补充剂组合物。
将本发明的组合物用作补充剂组合物时,可以直接摄取本发明的组合物,也可以在补充剂、饮料、食品、药物或饲料中添加本发明的组合物来摄取。
将本发明的组合物添加至补充剂、饮料、食品、药物或饲料中时,优选通过将本发明的组合物滴加在饮料、食品、药物或饲料中数滴而利用。
[制造方法]
本发明的组合物可以通过按任意的顺序将菌粉、脂肪油、以及选自由微粉和表面活性剂组成的组中的至少1种进行混合而制造。
本发明的组合物还可以通过在包含菌粉和脂肪油的悬浮液中混合选自由微粉和表面活性剂组成的组中的至少1种而制造。
混合的方法没有特别限定。例如,可以通过搅拌将菌粉、脂肪油、以及选自由微粉和表面活性剂组成的组中的至少1种进行混合。
组合物包含添加剂的情况,其混合顺序没有特别限定。可以包含在含有菌粉和脂肪油的悬浮液中,也可以在将选自由微粉和表面活性剂组成的组中的至少1种进行混合时进行添加。
[应用]
上述的微粉和表面活性剂可适宜用作抗结块剂。
实施例
以下通过实施例对本发明进行更具体地说明,但本发明不限定于后述的实施例。对于本发明,只要不改变其主旨就可以进行各种变形。
[菌粉的制备]
〈菌粉1〉
将婴儿双歧杆菌(Bifidobacterium infantis)NITE BP-02623接种于含有蛋白质、氨基酸和糖源的培养基中,在32~41℃下进行5~24小时培养后,通过离心分离从培养液中回收了菌体(湿菌体)。使用冷冻干燥机(共和真空技术株式会社制),进行18~96小时的冷冻干燥,将冷冻干燥结束后的菌体块物理地粉碎而得到冷冻干燥粉末。以1:3(w/w)的比例将得到的冷冻干燥粉末和木薯淀粉研制,得到双歧杆菌属细菌粉。
〈菌粉2〉
将短双歧杆菌(Bifidobacterium breve)NITE BP-02622接种于含有蛋白质、氨基酸和糖源的培养基中,在32~41℃下进行5~24小时培养后,通过离心分离从培养液中回收了菌体(湿菌体)。使用冷冻干燥机(共和真空技术株式会社制),进行120小时的冷冻干燥,将冷冻干燥结束后的菌体块物理地粉碎而得到冷冻干燥粉末。以1:3(w/w)的比例将得到的冷冻干燥粉末和木薯淀粉研制,得到双歧杆菌属细菌粉。
〈菌粉3〉
将长双歧杆菌长亚种(Bifidobacterium longum subsp.longum)NITE BP-02621接种于含有蛋白质、氨基酸和糖源的培养基中,在32~41℃下进行5~24小时培养后,通过离心分离从培养液中回收了菌体(湿菌体)。使用冷冻干燥机(共和真空技术株式会社制),进行18~96小时的冷冻干燥,将冷冻干燥结束后的菌体块物理地粉碎而得到冷冻干燥粉末。以1:3(w/w)的比例将得到的冷冻干燥粉末和玉米淀粉研制,得到双歧杆菌属细菌粉。
〈菌粉4〉
将长双歧杆菌长亚种(Bifidobacterium longum subsp.longum)NITE BP-02621接种于含有蛋白质、氨基酸和糖源的培养基中,在32~41℃下进行5~24小时培养后,通过离心分离从培养液中回收了菌体(湿菌体)。使用冷冻干燥机(共和真空技术株式会社制),进行18~96小时的冷冻干燥,将冷冻干燥结束后的菌体块物理地粉碎而得到冷冻干燥粉末。以1:3(w/w)的比例将得到的冷冻干燥粉末和马铃薯淀粉研制,得到双歧杆菌属细菌粉。
〈菌粉5〉
将加氏乳杆菌(Lactobacillus gasseri)NITE BP-01669接种于含有蛋白质、氨基酸和糖源的培养基中,在32~41℃下进行5~24小时培养后,通过离心分离从培养液中回收了菌体(湿菌体)。使用冷冻干燥机(共和真空技术株式会社制),进行18~96小时的冷冻干燥,将冷冻干燥结束后的菌体块物理地粉碎而得到冷冻干燥粉末。以1:3(w/w)的比例将得到的冷冻干燥粉末和玉米淀粉研制,得到乳酸菌粉。
〈菌粉6〉
将嗜酸乳杆菌(Lactobacillus acidophilus)NITE BP-01695接种于含有蛋白质、氨基酸和糖源的培养基中,在32~41℃下进行5~24小时培养后,通过离心分离从培养液中回收了菌体(湿菌体)。使用冷冻干燥机(共和真空技术株式会社制),进行18~96小时的冷冻干燥,将冷冻干燥结束后的菌体块物理地粉碎而得到冷冻干燥粉末。以1:3(w/w)的比例将得到的冷冻干燥粉末和玉米淀粉研制,得到乳酸菌粉。
〈菌粉7〉
将副干酪乳杆菌(Lactobacillus paracasei)NITE BP-01633接种于含有蛋白质、氨基酸和糖源的培养基中,在32~41℃下进行5~24小时培养后,通过离心分离从培养液中回收了菌体(湿菌体)。使用冷冻干燥机(共和真空技术株式会社制),进行18~96小时的冷冻干燥,将冷冻干燥结束后的菌体块物理地粉碎而得到冷冻干燥粉末。以1:3(w/w)的比例将得到的冷冻干燥粉末和麦芽糊精研制,得到乳酸菌粉。
〈菌粉8〉
将瑞士乳杆菌(Lactobacillus helveticus)NITE BP-01671接种于含有蛋白质、氨基酸和糖源的培养基中,在32~41℃下进行5~24小时培养后,通过离心分离从培养液中回收了菌体(湿菌体)。以1:4(固体成分换算比、w/w)的比例将浓缩菌液和淀粉分解物混合后,利用喷雾干燥器进行喷雾干燥,得到乳酸菌粉。
〈菌粉9〉
将长双歧杆菌婴儿亚种(Bifidobacterium longum subsp.infantis)NITE BP-02623接种于含有蛋白质、氨基酸和糖源的培养基中,在32~41℃下进行5~24小时培养后,通过离心分离从培养液中回收了菌体(湿菌体)。使用冷冻干燥机(共和真空技术株式会社制),进行18~96小时的冷冻干燥,将冷冻干燥结束后的菌块物理地粉碎而得到冷冻干燥粉末。
[分散性的评价方法]
将制得的组合物的10mL填充于玻璃制试管中,用橡胶栓进行密封。将填充有组合物的试管在设定于5℃的恒温箱中静置30天。将静置后的组合物以每秒1次左右的速度进行20次倾倒混合后,观察试管底面。
依据以下的基准,按A~E这5个等级评价了分散性。
A…底面未残留菌粉或载体。
B…与A相比,底面残留有较多的菌粉或载体。
C…与B相比,底面残留有较多的菌粉或载体。
D…与C相比,底面残留有较多的菌粉或载体。
E…难以悬浮。
[参考例]
将木薯淀粉(参考例1)或麦芽糊精(参考例2)2.5质量%与MCT油(S9013)97.5质量%混合,制备了100质量%的组合物。
使用制得的组合物,依据上述的评价方法评价了分散性。将评价结果示于表1。
[表1]
参考例1 | 参考例2 | |
评价 | C | B |
未配混菌粉的参考例1和参考例2中,载体的分散性优异。由此启示出,油滴中的菌粉的结块是由菌粉中包含的菌体本身所引起的。
[实施例和比较例]
<例A1~例A22>
例A1是比较例,例A2~例A22是实施例。
以表2或表3所示的比例(单位:质量%)将菌粉、微粉或表面活性剂、以及脂肪油进行混合,使用磁力搅拌器制成均匀的分散液。表面活性剂中,将B-100D添加至脂肪油中并在90℃的热水浴中使其溶解,恢复至室温后,添加菌粉并进行均匀地混合。
使用制得的组合物,依据上述的评价方法评价了分散性。将评价结果示于表2和表3。
[表2]
[表3]
配混了微粉或表面活性剂的例A2~例A22与未配混这些的例A1相比,菌粉的分散性优异。
配混了阴离子表面活性剂(硬脂酸钙)的例A8~例A10中,分散性的评价均为B,与例A1相比,菌粉的分散性良好。
配混了HLB值低于7的非离子表面活性剂(B-100D、S-170)的例A11~例A16与HLB值为7以上的非离子表面活性剂(S-770、S-1570)的配混了例A17~例A22相比,菌粉的分散性良好。
〈例B1~例B9〉
例B1是比较例,例B2~例B9是实施例。
以表4所示的比例(单位:质量%)将菌粉、微粉或表面活性剂、以及脂肪油进行混合,使用磁力搅拌器制成均匀的分散液。表面活性剂中,将L-195和POS-135添加至脂肪油中并在90℃的热水浴中使其溶解,恢复至室温后,添加菌粉并进行均匀地混合。
使用制得的组合物,依据上述的评价方法评价了分散性。将评价结果示于表4。
[表4]
配混了微粉或表面活性剂的例B2~例B9与未配混这些的例B1相比,菌粉的分散性优异。
配混了作为微粉的磷酸三钙的例B2、配混了作为阴离子表面活性剂的硬脂酸Mg的例B3、配混了作为非离子表面活性剂的B-370F(HLB值=3)的例B7、配混了作为非离子表面活性剂的POS-135(HLB值=1)的例B9中,分散性的评价为A,菌粉的分散性特别优异。
〈例C1~例C11〉
例C1是比较例,例C2~例C11是实施例。
以表5所示的比例(单位:质量%)将菌粉、微粉或表面活性剂、以及脂肪油进行混合,使用磁力搅拌器制成均匀的分散液。表面活性剂中,将B-100D添加至脂肪油中并在90℃的热水浴中使其溶解,恢复至室温后,添加菌粉并进行均匀地混合。
使用制得的组合物,依据上述的评价方法评价了分散性。将评价结果示于表5。
[表5]
配混了微粉或表面活性剂的例C2~例C11与未配混这些的例C1相比,菌粉的分散性优异。
配混了作为微粉的磷酸三钙的例C5、配混了作为非离子表面活性剂的B-100D(HLB值=3)的例C8、配混了作为非离子表面活性剂的B-370F(HLB值=3)的例C11中,分散性的评价为A,菌粉的分散性特别优异。
〈例D1~例D12〉
例D1、例D3、例D5、例D7、例D9和例D11是比较例,例D2、例D4、例D6、例D8、例D10和例D12是实施例。
以表6所示的比例(单位:质量%)将菌粉、微粉或表面活性剂、以及脂肪油进行混合,使用磁力搅拌器制成均匀的分散液。
使用制得的组合物,依据上述的评价方法评价了分散性。将评价结果示于表6。
[表6]
配混了硬脂酸钙的例D2、例D4、例D6、例D8、例D10和例D12中,分散性的评价为A或B,菌粉的分散性良好。
在使用了冷冻干燥菌粉的例(例D2、例D4、例D6、例D8、例D10)和使用了喷雾干燥菌粉的例(例D12)之间,未观察到添加硬脂酸钙时分散性的显著差异。
〈例E1、例E2〉
例E1是比较例,例E2是实施例。
以表7所示的比例(单位:质量%)将菌粉、微粉或表面活性剂、以及脂肪油进行混合,使用磁力搅拌器制成均匀的分散液。
使用制得的组合物,依据上述的评价方法评价了分散性。将评价结果示于表7。
[表7]
配混了硬脂酸钙的例E2中,分散性的评价为B,菌粉的分散性良好。
即使使用未混合研制剂的状态的菌粉,通过添加硬脂酸钙,分散性也得到了改善。
〈例F1、例F2〉
例F1是比较例,例F2是实施例。
直接使用市售的补充剂组合物(Baby Pro-Bio Bifidum M1,BEAN STALK SNOWCO.,LTD.制)(例F1)、或添加1.0质量%硬脂酸钙(例F2),制备了评价用组合物。
上述补充剂组合物是以双歧杆菌粉(动物双歧杆菌乳亚种BB-12:DSM 15954)、向日葵籽油、抗氧化剂(维生素E)、和柠檬酸作为原材料的组合物。
使用制得的评价用组合物,依据上述的评价方法评价了分散性。将评价结果示于表8。
[表8]
F1 | F2 | |
评价 | E | C |
即使是市售的补充剂组合物,通过添加硬脂酸钙,分散性也得到改善。
表2~表7中的术语的含义如下所述。
(菌粉)
菌粉1…如上所述制备的菌粉1
菌粉2…如上所述制备的菌粉2
菌粉3…如上所述制备的菌粉3
菌粉4…如上所述制备的菌粉4
菌粉5…如上所述制备的菌粉5
菌粉6…如上所述制备的菌粉6
菌粉7…如上所述制备的菌粉7
菌粉8…如上所述制备的菌粉8
菌粉9…如上所述制备的菌粉9
(脂肪油)
S9013中链脂肪酸甘油三酯(MCT油S9013、太阳油脂株式会社制)
(微粉)
微晶纤维素…Ceolus FD-F20(旭化成株式会社制)
磷酸三Ca…磷酸三钙(太平化学产业株式会社制)
微粒二氧化硅…SYLOPAGE720(FUJI SILYSIA CHEMICAL LTD.制)
(表面活性剂)
硬脂酸Ca…硬脂酸钙(太平化学产业株式会社制)
硬脂酸Mg…硬脂酸镁(San-Ei Gen F.F.I.,Inc.制)
B-100D…甘油脂肪酸酯(Ryoto Polyglycerol Ester B-100D、Mitsubishi-Chemical Foods Corporation制;HLB值3)
S-170…蔗糖脂肪酸酯(Ryoto Sugar Ester S-170、Mitsubishi-Chemical FoodsCorporation制;HLB值1)
P-170…蔗糖脂肪酸酯(Ryoto Sugar Ester P-170、Mitsubishi-Chemical FoodsCorporation制;HLB值1)
O-170…蔗糖脂肪酸酯(Ryoto Sugar Ester O-170、Mitsubishi-Chemical FoodsCorporation制;HLB值1)
L-195…蔗糖脂肪酸酯(Ryoto Sugar Ester L-195、Mitsubishi-Chemical FoodsCorporation制;HLB值1)
B-370F…蔗糖脂肪酸酯(Ryoto Sugar Ester B-370F、Mitsubishi-ChemicalFoods Corporation制;HLB值3)
ER-190…蔗糖脂肪酸酯(Ryoto Sugar Ester ER-190、Mitsubishi-ChemicalFoods Corporation制;HLB值1)
POS-135…蔗糖脂肪酸酯(Ryoto Sugar Ester POS-135、Mitsubishi-ChemicalFoods Corporation制;HLB值1)
S-770…蔗糖脂肪酸酯(Ryoto Sugar Ester S-770、Mitsubishi-Chemical FoodsCorporation制;HLB值7)
S-1570蔗糖脂肪酸酯(Ryoto Sugar Ester S-1670、Mitsubishi-Chemical FoodsCorporation制;HLB值16)
[结果的说明]
实施例中,观察到抑制菌粉的沉淀和固化、以及结块的效果。
配混了表面活性剂时,有显示出优异的效果的倾向。
需要说明的是,配混了菌粉的组合物与仅配混了载体的组合物相比时,确认到配混了菌粉的组合物倾向于更多地残留于底面,因此强烈需要配混菌粉的组合物以消除沉淀/固化、及结块。
产业上的可利用性
本发明的组合物可以作为补充剂组合物直接摄取、或添加在食品中加以利用。
Claims (10)
1.选自由微粉和表面活性剂组成的组中的至少1种作为菌粉用抗结块剂在包含脂肪油的组合物中的应用。
2.根据权利要求1所述的应用,其中,所述组合物为悬浮液。
3.根据权利要求1或2所述的应用,其中,所述菌粉包含选自由乳酸菌的活菌、乳酸菌的死菌、双歧杆菌属细菌的活菌和双歧杆菌属细菌的死菌组成的组中的至少1种。
4.根据权利要求1或2所述的应用,其中,所述微粉包含选自由微晶纤维素和磷酸三钙组成的组中的至少1种。
5.根据权利要求1或2所述的应用,其中,所述微粉为无机物。
6.根据权利要求5所述的应用,其中,所述无机物为磷酸三钙。
7.根据权利要求1或2所述的应用,其中,所述表面活性剂包含选自由阴离子表面活性剂和HLB值低于7的非离子表面活性剂组成的组中的至少1种。
8.根据权利要求7所述的应用,其中,所述阴离子表面活性剂包含选自由硬脂酸金属盐组成的组中的至少1种,所述HLB值低于7的非离子表面活性剂包含选自由HLB值低于7的蔗糖脂肪酸酯和HLB值低于7的甘油脂肪酸酯组成的组中的至少1种。
9.根据权利要求1或2所述的应用,其中,相对于所述组合物的总质量,包含0.5~10质量%的所述菌粉。
10.根据权利要求1或2所述的应用,其中,所述组合物还包含添加剂。
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JP2017112995A (ja) * | 2015-12-18 | 2017-06-29 | アサヒ飲料株式会社 | 微生物菌体含有飲料 |
TW201906538A (zh) * | 2017-06-14 | 2019-02-16 | 日商朝日飲料股份有限公司 | 含有微生物菌體之碳酸飲料、及提高碳酸飲料中微生物菌體粉末之沉澱物或凝聚物之分散性之方法 |
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