CN114208939A - 一种宠物犬猫用植物基酸奶及其制备方法 - Google Patents
一种宠物犬猫用植物基酸奶及其制备方法 Download PDFInfo
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Abstract
本发明涉及益生菌应用技术领域,特别涉及一种宠物犬猫用植物基酸奶及其制备方法。所述植物基酸奶是利用凝结芽孢杆菌VHProbi C08发酵制备得到的,酸度适宜,活菌数高,风味独特。经代谢组学分析可知,植物基酸奶中的氨基酸、核苷酸、维生素等初级代谢物种类丰富,含量高,有助于改善酸奶的风味质地,提高酸奶的营养益生作用,促进宠物犬猫健康成长,具有极大的市场潜力。
Description
技术领域
本发明涉及益生菌应用技术领域,特别涉及一种宠物犬猫用植物基酸奶及其制备方法。
背景技术
植物基酸奶是以大豆、巴旦木、鹰嘴豆等植物原料为基底,通过添加白砂糖、淀粉等其他辅助原料,经乳酸菌菌株发酵后得到的具有特殊香气的产品。有数据显示,2020年,美国市场植物性酸奶产品增长20.2%,几乎是传统酸奶的7倍,销售额达3.43亿美元。在国内,很多大品牌也开始了植物基酸奶的布局,未来植物基酸奶市场发展空间广阔。
消费者对健康的关注是推动植物基酸奶发展起来的主要原因,这是因为植物基酸奶相较于采用牛乳、羊乳发酵的传统酸奶具有较多优势。很多科学研究表明植物蛋白可提供比动物蛋白质更多的营养价值,且已经将动物脂肪或蛋白的消耗与慢性疾病相关联。植物基酸奶因不采用牛乳,更适合牛乳过敏、乳糖不耐受、崇尚素食主义或关心环境伦理的消费者饮用。植物基酸奶的供应柔和性更强,产量基本保持稳定,植物原料的运输与保存也更便捷。植物基酸奶不含胆固醇和乳糖,产品质构特性与动物乳基酸奶基本相同,在健康和实现资源可持续方面是普通酸奶的理想替代品。
宠物犬猫的体温和胃肠道pH与人体存在差别。成年犬体温37.5~38.5℃、幼犬38.5~39℃,猫的正常体温为38~38.5℃,均高于人体正常体温。胃分泌的酸是盐酸,有2种存在形式:一种为游离酸,另一种为结合酸,即与蛋白质结合的盐酸蛋白酸。两种酸合在一起称为总酸。在纯胃液中绝大部分是游离酸,pH为0.9~1.5。人和普通动物的胃酸成分基本上没大区别,只是强弱的区别,但是狗的胃酸比人强几百倍,可以消化骨头。宠物犬猫体内普遍缺少乳糖酶,易乳糖不耐。但是目前市面上专为宠物犬猫设计的酸奶种类很少,且均为牛乳、羊乳发酵的传统酸奶,给犬猫喂食传统酸奶或人用酸牛奶,犬猫难以消化吸收,会给肠胃造成很大的负担。
植物基酸奶的营养价值在于不含或含少量过敏原、无乳、无动物源性成分、不含或含少量乳糖等,植物基酸奶因不采用牛乳,更适合体内普遍缺少乳糖酶、易乳糖不耐的宠物犬猫饮用。不同植物蛋白适用的发酵菌种不同,因此需根据所用原料挑选合适的菌种。目前植物基酸奶大多以大豆为基底进行发酵,发酵菌种的选择对于发酵风味、粘度、货架期稳定性等有重要影响。因此,为解决上述问题,本研究将开发一款风味独特、营养丰富、能更好地适应宠物犬猫胃肠道环境、更适合宠物犬猫饮用的植物基酸奶。
植物基酸奶的营养价值在于不含或含少量过敏原、无乳、无动物源性成分、不含或含少量乳糖等,植物基酸奶因不采用牛乳,更适合体内普遍缺少乳糖酶、易乳糖不耐的宠物犬猫饮用。植物基酸奶的供应柔和性更强。植物作为酸奶原料的稳定性更强,对环境也更加友好。植物原料的运输与保存也更便捷,产量基本保持稳定。凝结芽孢杆菌比普通发酵菌种能更好地适应宠物犬猫胃肠道环境,从而帮助调节宠物肠道微生态平衡,促进消化吸收。因此本研究将其应用于宠物植物基酸奶发酵,以获得更适合宠物犬猫饮用的酸奶。
发明内容
本发明为解决现有技术问题,提供了一种新型宠物犬猫用植物基酸奶及其制备方法。所述酸奶中活菌量高,酸度适宜,代谢产物种类丰富,营养价值高,市场前景广阔。
本发明一方面涉及一种植物基酸奶,是以凝结芽孢杆菌对原料进行发酵制备得到的。
所述凝结芽孢杆菌为凝结芽孢杆菌 VHProbi C08(Bacillus coagulans VHProbiC08),已于 2019 年 9 月 23 日保藏于中国武汉大学的中国典型培养物保藏中心,其保藏号为 CCTCC NO:M2019738。
所述原料的组分及其重量份数分别为:植物蛋白粉3~10份、低聚果糖4~10份、L-赖氨酸0.5~1.5份、乳酸钙0.3~0.6份、牛磺酸0.02~0.04份、黄原胶0.2~0.5份、纯净水80~100份。
进一步优选的,所述原料的组分及其重量份数分别为:植物蛋白粉4份、低聚果糖8份、L-赖氨酸1份、乳酸钙0.4份、牛磺酸0.03份、黄原胶0.35份、纯净水90份。
所述植物蛋白粉选自豆粉、燕麦粉、荞麦粉、青稞粉中的任意一种或两种或多种的组合。
所述植物蛋白粉优选豆粉。
本发明一方面涉及一种植物基酸奶的制备方法,包括如下步骤:
(1)按比例称取各原料组分,充分混匀,均质,得到发酵培养基,其中各组分及其重量份数分别为植物蛋白粉3~10份、低聚果糖4~10份、L-赖氨酸0.5~1.5份、乳酸钙0.3~0.6份、牛磺酸0.02~0.04份、黄原胶0.2~0.5份、纯净水80~100份;
(2)将均质后的发酵培养基在115~121℃条件下高温灭菌;
(3)待发酵培养基冷却至37~45℃时,接种活化后的凝结芽孢杆菌VHProbi C08菌液,至发酵培养基中初始活菌量为106~107CFU/mL,37~45℃条件下发酵14~18h;
(4)在4℃条件下酸化12h,即制得植物基酸奶。
上述制备方法中步骤(1)中所述发酵培养液的组分及其重量份数分别为豆粉4份、低聚果糖8份、L-赖氨酸1份、乳酸钙0.4份、牛磺酸0.03份、黄原胶0.35份、纯净水90份。
本发明还提供了所述植物基酸奶在制备宠物犬猫食品中的应用。
本发明选用凝结芽孢杆菌VHProbi C08发酵制备植物基酸奶。该菌株具有耐高温、耐酸、耐胆盐、易培养、易保存和促进肠道双歧杆菌等益生菌的生长繁殖的特点,其能成功在胃肠环境下繁殖并形成优势菌群,而且还具有抗腹泻、降解胆固醇和降血糖的益生功效。
本发明制备得到的植物基酸奶pH和酸度适宜,粘度和持水力较好,具有良好的质地和口感,且酸奶中活菌量高于4.0×108CFU/mL,粗蛋白含量高于国家标准,总糖含量较市面中其他酸奶制品偏少,有助于预防宠物犬猫的龋齿和肥胖。
所述植物基酸奶中共检测出1113种代谢产物。与发酵前相比,植物基酸奶中差异显著的代谢物共有9类,156 种,占总代谢物的 14.02%,其中氨基酸及其衍生物、有机杂环化合物、有机氧化合物、脂质、核苷酸及其衍生物等 5 类代谢物含量最高,分别占比为29.14%、19.21%、13.35%、12.58%、6.62%。与发酵前相比,植物基酸奶中含量显著提高的代谢物为胞苷、L-赖氨酸、苯乙酰甘氨酸、黄嘌呤核苷、D-葡萄糖酸等共14 种。酸奶中富含有L-赖氨酸、L-天冬氨酸、L-色氨酸等16种氨基酸及其衍生物,以及吡哆胺、D-泛酸、烟酰胺和D-脱硫生物素等维生素和辅酶因子等等,代谢产物种类非常丰富,并且代谢产物中的多种营养益生物质,如L-天冬氨酸、可的宁N-葡萄糖醛酸苷、没食子酸等代谢物的含量较发酵前提高了几十倍甚至100多倍,极大地提高了植物基酸奶的营养价值。
本发明制备的植物基酸奶营养丰富,风味独特,适口性与耐口性俱佳,有良好的市场潜力和应用前景。
附图说明
图1为总离子色谱图,其中a为质控样本的正离子模式总离子流图,b为质控样本的负离子模式总离子流图;
图2为植物基酸奶发酵前后的OPLS-DA 图和S-plot 图;其中a为OPLS-DA 图,b为S-plot 图;
图3为植物基酸奶中差异显著的代谢物KEGG通路富集图。
具体实施方式
本发明实施例中选用的凝结芽孢杆菌 VHProbi C08(Bacillus coagulansVHProbi C08)筛选自酸白菜样品,已于 2019 年 9 月 23 日保藏于中国武汉大学的中国典型培养物保藏中心,其保藏号为 CCTCC NO:M2019738。
凝结芽孢杆菌 VHProbi C08 不产生溶血素,不会溶解血细胞,对常见抗生素敏感,生物安全性良好;能够耐受较高的盐度,最大耐受盐浓度为 7%;能在较广阔的温度范围内生长,并且能产生耐热性很强的孢子,方便该益生菌株后期工业化加工;体外细胞表面疏水性为 8.58%,对肠道细胞具有粘附性。
凝结芽孢杆菌 VHProbi C08 具备较强的抗氧化功能,其中对 DPPH 自由基清除率为 87.80%,HRS 自由基清除率为 47.79%;上清液抗脂质过氧化抑制率为 84.46%,菌体抗脂质过氧化抑制率为 44.88%。
凝结芽孢杆菌 VHProbi C08 具备较强的降解胆固醇能力,其中体外胆固醇降解率达到 39.21%;在高血脂小鼠模型中,用本发明提供的凝结芽孢杆菌VHProbi C08 喂食模型鼠 30 天后预处理组血液胆固醇水平降低了约 34%,后处理组血液胆固醇水平降低了约10%;40 天后,凝结芽孢杆菌 VHProbi C08预处理组血液胆固醇水平降低了约 39%,后处理组血液胆固醇水平降低了约30%,效果非常显著。
凝结芽孢杆菌 VHProbi C08 能有效预防和缓解腹泻,抑制肠蠕动。给大鼠连续服用凝结芽孢杆菌 VHProbi C08 7 天后,能有效延迟腹泻时间,减少粪便量。凝结芽孢杆菌VHProbi C08 代谢产生的有机酸等代谢产物能够抑制有害菌的生长,促进肠道对营养物质的消化和吸收,恢复肠道的微生物屏障功能,刺激机体免疫功能,缩短疾病病程。
下面结合具体实施例,对本发明做进一步说明,但本发明不受实施例的限制。
实施例1
一种植物基酸奶,是通过如下方法制备得到的:
(1)按比例称取各原料组分,充分混匀,均质,得到发酵培养基,其中各组分及其重量份数分别为豆粉10份、低聚果糖4份、L-赖氨酸0.5份、乳酸钙0.3份、牛磺酸0.04份、黄原胶0.2份、纯净水100份;
(2)将均质后的发酵培养基在115~121℃条件下高温灭菌;
(3)待发酵培养基冷却至40℃时,接种活化后的凝结芽孢杆菌VHProbi C08,至发酵培养基中活菌量为3.5×106CFU/mL,40℃条件下发酵24h;
(4)在4℃条件下酸化12h,即制得植物基酸奶。
实施例2
一种植物基酸奶,是通过如下方法制备得到的:
(1)按比例称取各原料组分,充分混匀,均质,得到发酵培养基,其中各组分及其重量份数分别为豆粉3份、低聚果糖10份、L-赖氨酸1.5份、乳酸钙0.6份、牛磺酸0.02份、黄原胶0.5份、纯净水80份;
(2)将均质后的发酵培养基在115~121℃条件下高温灭菌;
(3)待发酵培养基冷却至45℃时,接种活化后的凝结芽孢杆菌VHProbi C08,至发酵培养基中活菌量为106CFU/mL,45℃条件下发酵18h;
(4)在4℃条件下酸化12h,即制得植物基酸奶。
实施例3
一种植物基酸奶,是通过如下方法制备得到的:
(1)按比例称取各原料组分,充分混匀,均质,得到发酵培养基,其中各组分及其重量份数分别为豆粉4份、低聚果糖8份、L-赖氨酸1份、乳酸钙0.4份、牛磺酸0.03份、黄原胶0.35份、纯净水90份;
(2)将均质后的发酵培养基在115~121℃条件下高温灭菌;
(3)待发酵培养基冷却至37℃时,接种活化后的凝结芽孢杆菌VHProbi C08,至发酵培养基中活菌量为107CFU/mL,37℃条件下发酵14h;
(4)在4℃条件下酸化12h,即制得植物基酸奶。
实施例4植物基酸奶活菌数以及粗蛋白和总糖含量的测定
分别对本发明实施例1~3制备得到的植物基酸奶样品中活菌数以及粗蛋白和总糖含量进行测定。
1、活菌数的测定方法:
用葡萄糖酵母提取物BC琼脂培养基(1L)进行凝结芽孢杆菌活菌数的测定,其配方如下:酵母粉5.0g,蛋白胨5.0g,葡萄糖5.0g,K2HPO4 0.5g,KH2PO4 0.5g,MgSO4 0.3g,微量元素溶液1.0mL,琼脂粉15.0g,蒸馏水1000mL ,调pH6.3,121℃灭菌15min。
微量元素溶液(mg/mL):NaCl 10g,7H2O·FeSO4 18g,H2O·MnSO4 16g,7H2O·ZnSO4 1.6g,5H2O·CuSO4 1.6g,7H2O·CoSO4 1.6g,过滤除菌。
取少量样品进行10倍梯度稀释,直到最后的稀释预计含有大约30个菌落形成单位(CFU)/mL。在10~20分钟内稀释样品,选取最后三个稀释剂用于分析。每个稀释度吸取1mL样品匀液于无菌平皿内,将冷却至48 ℃的BC琼脂培养基倾注入平皿约15mL,转动平皿使混合均匀。40℃±2℃厌氧培养48h。孵育48小时后,计数培养皿上的菌落。计数含有30到300个菌落的平板,只计算以下出现的菌落:在BC琼脂培养基表面菌落直径应在1 mm ~ 5 mm之间,白色至乳白色,凸出,边缘完整,表面光滑;在BC琼脂培养基内部的菌落直径应为0.5 mm~1mm,呈奶油色针尖状。待长出单菌落后进行计数。
2、粗蛋白含量的测定方法:参照GB5009.5-2016《食品安全国家标准 食品中蛋白质的测定》中自动凯氏定氮仪法,将消化后的样品在自动凯氏定氮仪上实现自动加液、蒸馏、滴定和记录滴定数据的过程,根据公式(B)计算样品中蛋白质的含量,结果如表1所示。
其中X为试样中蛋白质的含量(g/100g),V1为试液消耗硫酸标准滴定液的体积(mL),V2为试剂空白消耗硫酸标准滴定液的体积(mL),c为硫酸标准滴定溶液浓度(mol/L),m为试样的质量(g),V3为吸取消化液的体积(mL),F为氮换算为蛋白质的系数。
3、总糖含量的测定方法:准确称取0.5g植物基酸奶样品,放在大试管中,加入6MHCL 10 mL,在沸水浴中加热0.5h,冷却至室温后加入1滴酚酞指示剂,以6M NaOH溶液中和至溶液呈微红色,将试管中的溶液转移至50 mL容量瓶,并用去离子水多次润洗试管定容至50 mL;用DNS法测定还原糖含量,即为总糖含量,结果如表1所示。
表1 植物基酸奶中的活菌数以及粗蛋白和总糖含量
植物基酸奶 | 活菌数(CFU/mL) | 总糖(mg/ mL) | 粗蛋白(%) |
实施例1 | 4.45×10<sup>8</sup>±0.06 | 38.26±0.04 | 3.08±0.03 |
实施例2 | 4.62×10<sup>8</sup>±0.08 | 40.15±0.06 | 3.21±0.01 |
实施例3 | 4.75×10<sup>8</sup>±0.02 | 40.84±0.04 | 3.26±0.07 |
由表1的数据可以看出,本发明制备得到的植物基酸奶中活菌量较高,粗蛋白含量高于GB19302-2010《食品安全国家标准发酵乳》中发酵乳蛋白质含量≥2.9%的标准,并且总糖含量较市面中其他酸奶制品偏少,有利于预防宠物犬猫的龋齿和肥胖。其中,实施例3制备得到的植物基酸奶中活菌数、粗蛋白含量比实施例1和2相对较高。
实施例5植物基酸奶基本理化指标测定
对本发明实施例1~3制备得到的植物基酸奶进行基本理化指标的测定。
1、pH的测定:
取出酸奶样品10mL于相应的小烧杯中,校准pH计后,开始样品pH的测定,待示数稳定后读数并做好记录,每个样品读数3次,取平均值。
2、酸度的测定:
参照GB/T 12456-2008《食品中总酸的测定》测定滴定酸度,配制0.1mol/L的氢氧化钠标准溶液,称取10g酸奶样品进行3倍稀释处理,然后进行滴定,记录消耗的NaOH溶液体积V mL,根据酸度计算公式°T=[c(NaOH)/0.1]×V×10计算样品酸度。
3、粘度的测定:
用Brookfield粘度计进行酸奶粘度的测定,将酸奶样品加入样品杯中,再将转子浸入样品中合适位置,连接转子,设置转子和转速,当读数稳定后进行读数,每个样品测试3组数据,取平均值。
4、持水力的测定:
取酸奶样品10mL进行3000r/min离心10min处理,分离出相应的乳清,称量并记录酸奶去乳清前和去乳清后的质量,两者比值即为酸奶样品的持水力。
上述各项理化指标检测结果见表2。
表2 植物基酸奶基本理化指标
植物基酸奶 | pH | 酸度(°T) | 粘度(mPa﹒s) | 持水力(%) |
实施例1 | 4.66±0.02 | 82.08±0.11 | 287.0±0.09 | 61.45±0.12 |
实施例2 | 4.59±0.05 | 84.12±0.03 | 304.0±0.04 | 63.94±0.09 |
实施例3 | 4.56±0.09 | 85.41±0.07 | 309.0±0.06 | 65.55±0.07 |
由表2的数据可以看出,本发明实施例1~3制备得到的植物基酸奶pH和酸度均比较适宜,粘度和持水力较好,具有良好的质地和口感。
实施例6植物基酸奶代谢产物分析
申请人利用非靶向代谢组技术对本发明制备得到的植物基酸奶进行代谢产物进行分析,从而有助于了解凝结芽孢杆菌VHProbi C08在大豆植物基中的代谢特征,为植物基酸奶加工工艺改良、滋味品质提升以及功能性成分富集等方面研究提供依据。
6.1 试验方法
6.1.1 样本处理
精确移取实施例3制备得到的酸奶200µL至1.5 mL离心管中,加入800 µL 提取液(甲醇:乙腈=1:1(v:v)),含0.02 mg/mL的内标(L-2-氯苯丙氨酸);涡旋混匀30s后,低温超声提取30 min(5℃,40KHz);将样品静置于-20℃,30 min;离心15 min(13000g,4℃),移取上清液,氮气吹干;加入 100 μL复溶液(乙腈:水=1:1)复溶;涡旋混匀30s,低温超声萃取 5min(5℃,40 KHz);离心10 min(13000 g,4℃),移取上清液至带内插管的进样小瓶中上机分析;另外,每个样本分别移取等体积上清液混合后作为质控样本。
6.1.2 LC-MS检测
采用超高效液相色谱串联傅里叶变换质谱UHPLC-Q Exactive HF-X 系统,ACQUITY UPLC HSS T3色谱柱对样品进行分离。流动相A为95%水+5%乙腈(含0.1%甲酸),流动相B为47.5%乙腈+47.5%异丙醇+5%水(含0.1%甲酸),进样量为2μL,柱温为40℃,梯度洗脱程序见表3。
表3 流动相洗脱梯度
Time(min) | Flow rate(mL/min) | A(%) | B(%) |
0 | 0.4 | 100 | 0 |
3.5 | 0.4 | 75.5 | 24.5 |
5 | 0.4 | 35 | 65 |
5.5 | 0.4 | 0 | 100 |
7.4 | 0.6 | 0 | 100 |
7.6 | 0.6 | 48.5 | 51.5 |
7.8 | 0.5 | 100 | 0 |
9 | 0.4 | 100 | 0 |
10 | 0.4 | 100 | 0 |
质谱条件:样品经电喷雾电离,分别采用正、负离子扫描模式采集质谱信号。具体参数如表4所示。
样品经色谱分离流出的组分不断进入质谱,质谱连续扫描进行数据采集,每一次扫描得到一张质谱图,将每一张质谱图中所有离子强度相加,得到一个总的离子流强度。以时间为横坐标、离子强度总和为纵坐标绘制得到总离子色谱图。通过总离子色谱图评价仪器误差及数据可靠性。
表4 质谱参数
描述 | Description | 参数 |
扫描范围 | Scan typr(m/z) | 70-1050 |
鞘气流速 | Shesth gas flow rate (arb) | 50 |
辅助气流速 | Aux gas flow rate (arb) | 13 |
加热温度 | Heater temp (℃) | 425 |
毛细管温度 | Capillary temp (℃) | 325 |
喷雾电压(正模式) | Spray voltage (+) (V) | 3500 |
喷雾电压(负模式) | Spray voltage (-) (V) | -3500 |
S-Lens电压 | S-Lens RF Leval | 50 |
碰撞能 | Normalized collision energy (eV) | 20,40,60 |
分辨率(Full MS) | Resolution (Full MS<sup>)</sup> | 60000 |
分辨率(Full MS<sup>2</sup>) | Resolution (MS<sup>2)</sup> | 7500 |
6.1.3 数据处理与分析方法
将质谱检测获得的原始数据导入代谢组学处理软件 ProgenesisQI(WatersCorporation, Milford, USA)进行基线过滤、峰识别、积分、保留时间校正、峰对齐等,最终得到含保留时间、质荷比和峰强度等信息的数据矩阵。其后采用该软件进行特征峰搜库鉴定,将 MS 和 MS/MS 质谱信息与代谢数据库进行匹配,MS 质量误差设置为小于 10ppm,同时根据二级质谱匹配得分鉴定代谢物。主要数据库为 http://www.hmdb.ca/、https://metlin.scripps.edu/等公共数据库。比较发酵前后酸奶中代谢成分的差异变化,采用 OPLS-DA模型的变量投影重要性 (VIP, Variable importance in the projection)大于 1 和单维统计分析(正态分布数据采用双尾学生氏 t 检验和非正态分布数据采用Wilcoxon Mann-Whitney 检验)的 P值小于 0.05 相结合的方法筛选差异性代谢物,并结合 KEGG 数据库进行代谢通路分析。
6.2 结果与分析
6.2.1植物基酸奶总离子色谱图
植物基酸奶总离子色谱图如图1所示,a和b分别展示了正、负离子模式下,质控样品的总离子色谱图。由图3可见,该检测条件下,峰形良好,分布相对均匀;各色谱峰的响应强度和保留时间基本重叠,说明仪器误差较小,数据可靠。
6.2.2植物基酸奶OPLS-DA 分析
为了准确获得植物基酸奶发酵前后样品中的差异代谢物,采用OPLS-DA(orthogonal partial least squares discriminant analysis, OPLS-DA)有监督的判别方法对发酵前后的植物基酸奶样品进行分析,得到 OPLS-DA 得分图2。由图2可知,正离子模式下,植物基酸奶发酵前后的代谢产物明显分离,表明各组之间存在显著差异,全部样品组都位于置信区间内,同时参数 R2Y(cum)=1,Q2(cum)=0.994,均接近1,说明模型稳定,数据可靠。在 S-plot 图中,每个小点代表1 个变量,离原点越远的变量贡献越大,差异越明显,越有可能成为潜在生物标记物。图2中黑色的点表示这些代谢物的 VIP 值≥1,这些物质将进一步结合差异倍数和显著性差异参数进行再次筛选。
6.2.3植物基酸奶中差异代谢物鉴定与分析
检测分析结果显示,本发明提供的植物基酸奶中共检测出1113种代谢产物。与发酵前相比,植物基酸奶中差异显著的代谢物共有9类,156 种,占总代谢物的 14.02%,其中氨基酸及其衍生物、有机杂环化合物、有机氧化合物、脂质、核苷酸及其衍生物等 5 类代谢物含量最高,分别占比为 29.14%、19.21%、13.35%、12.58%、6.62%。
通过进一步比较发酵前后植物基酸奶中代谢成分的差异倍数变化可知:与发酵前相比,植物基酸奶中含量显著提高的代谢物共有 14 种,主要涉及氨基酸及其衍生物4种、核苷酸及其衍生物3种、有机杂环化合物3种、有机氧化合物1种、脂质1种、苯及其衍生物1种、苯丙烷和聚酮化合物1种。14种具体的代谢物如表 5 所示。
表5 发酵后含量显著提高的代谢物
化合物分类 | 分子式 | 代谢物名称 |
氨基酸及其衍生物 | C<sub>8</sub>H<sub>18</sub>N<sub>2</sub>O<sub>2</sub> | N<sub>6</sub>, N<sub>6</sub>-二甲基-L-赖氨酸 |
氨基酸及其衍生物 | C<sub>10</sub>H<sub>11</sub>NO<sub>3</sub> | 苯乙酰甘氨酸 |
氨基酸及其衍生物 | C<sub>4</sub>H<sub>7</sub>NO<sub>4</sub> | L-天冬氨酸 |
氨基酸及其衍生物 | C<sub>11</sub>H<sub>12</sub>N<sub>2</sub>O<sub>2</sub> | L-色氨酸 |
核苷酸及其衍生物 | C<sub>9</sub>H<sub>13</sub>N<sub>3</sub>O<sub>5</sub> | 胞苷 |
核苷酸及其衍生物 | C<sub>16</sub>H<sub>20</sub>N<sub>2</sub>O<sub>7</sub> | 可的宁N-葡萄糖醛酸苷 |
核苷酸及其衍生物 | C<sub>10</sub>H<sub>12</sub>N<sub>4</sub>O<sub>6</sub> | 黄嘌呤核苷 |
有机杂环化合物 | C<sub>9</sub>H<sub>9</sub>NO<sub>4</sub> | 无色多巴色素 |
有机杂环化合物 | C<sub>10</sub>H<sub>12</sub>N<sub>2</sub>O | 5-羟色胺 |
有机杂环化合物 | C<sub>8</sub>H<sub>7</sub>NO<sub>5</sub> | 5-羟基-6-甲基-3,4-吡啶二甲酸 |
有机氧化合物 | C<sub>6</sub>H<sub>10</sub>O<sub>8</sub> | D-葡萄糖酸 |
脂质 | C<sub>19</sub>H<sub>26</sub>O<sub>4</sub> | 泛醌-2 |
苯及其衍生物 | C<sub>7</sub>H<sub>6</sub>O<sub>5</sub> | 没食子酸 |
苯丙烷和聚酮化合物 | C<sub>9</sub>H<sub>8</sub>O<sub>3</sub> | 2-羟基肉桂酸 |
氨基酸是构建细胞、修复组织的基础材料,是机体生长发育所必需的营养物质,也是调节代谢,增加抵抗力所需物质.
核苷酸具有抗氧化作用,食物中缺乏核苷酸可损害肝脏、心脏、肠道和免疫系统。
有机杂环化合物是有机化合物中庞大的一类化合物,广泛存在于核酸、某些维生素、激素和生物碱等具有生物活性的分子中,在生物体内常常扮演着重要角色.
有机氧化合物主要包括碳水化合物和多元醇类化合物。碳水化合物是生命细胞结构的主要成分及主要供能物质,并且有调节细胞活动的重要功能;多元醇类化合物,如莽草酸、奎尼酸,可应用于抗菌、抗肿瘤、抗血栓形成和抗脑缺血等多种疾病的治疗,还具有抗炎、镇痛作用。
脂质具有抗氧化、抗癌、降低胆固醇、减肥、降血糖血脂等多种生理功能。
没食子酸具有抗炎、抗突变、抗氧化、抗自由基等多种生物学活性.
羟基肉桂酸可通过调节肠道菌群,缓解动物模型的炎症性肠炎、肠道缺血再灌注损伤、结直肠癌等疾病。
综上,本发明提供的植物基酸奶中富含有L-赖氨酸、L-天冬氨酸、L-色氨酸等16种氨基酸及其衍生物,以及吡哆胺、D-泛酸、烟酰胺和D-脱硫生物素等维生素和辅酶因子等等,代谢产物种类非常丰富,并且代谢产物中的多种营养益生物质,如L-天冬氨酸、可的宁N-葡萄糖醛酸苷、没食子酸等代谢物的含量较发酵前提高了几十倍甚至100多倍,极大地提高了植物基酸奶的营养价值。
6.2.4 凝结芽孢杆菌VHProbi C08在大豆植物基中的代谢通路分析
通路富集分析共获得72 种差异代谢通路,其中 19个通路存在显著差异(P<0.05)。排在前10 的代谢通路见图3。
通过对差异代谢物数量富集情况进行分析可知,本发明所述凝结芽孢杆菌VHProbi C08在大豆植物基中的主要代谢通路分别是①ABC转运蛋白(ABC transporters),主要包括L-苯丙氨酸、L-天冬氨酸和胞苷等20种代谢产物;②色氨酸代谢(Tryptophanmetabolism),主要包括5-羟色胺、L-色氨酸等14种代谢产物;③氨酰-tRNA生物合成(Aminoacyl-tRNA biosynthesis),主要包括L-天冬氨酸、L-谷氨酰胺和L-异亮氨酸等13种代谢产物;④氨基苯甲酸降解(Aminobenzoate degradation),主要包括没食子酸、丁香酸、香兰素等12种代谢产物;⑤酪氨酸代谢(tyrosine metabolism),主要包括L-酪氨酸、4-羟基苯基丙酮酸、无色多巴色素等11种代谢产物。
综上,本发明利用凝结芽孢杆菌VHProbi C08发酵制备得到的植物基酸奶酸度适宜,活菌数高,风味独特。经代谢组学分析可知,植物基酸奶中的氨基酸、核苷酸、维生素等初级代谢物种类丰富,含量高,有助于改善酸奶的风味质地,提高酸奶的营养益生作用,促进宠物犬猫健康成长,具有极大的市场潜力。
Claims (9)
1.一种植物基酸奶,其特征在于,所述的植物基酸奶是以凝结芽孢杆菌对原料进行发酵制备得到的。
2.如权利要求1所述的植物基酸奶,其特征在于,所述的凝结芽孢杆菌为凝结芽孢杆菌VHProbi C08(Bacillus coagulans VHProbi C08),保藏号为 CCTCC NO:M2019738。
3.如权利要求1或2所述的植物基酸奶,其特征在于,所述原料的组分及其重量份数分别为:植物蛋白粉3~10份、低聚果糖4~10份、L-赖氨酸0.5~1.5份、乳酸钙0.3~0.6份、牛磺酸0.02~0.04份、黄原胶0.2~0.5份、纯净水80~100份。
4.如权利要求3所述的植物基酸奶,其特征在于,所述原料的组分及其重量份数分别为:植物蛋白粉4份、低聚果糖8份、L-赖氨酸1份、乳酸钙0.4份、牛磺酸0.03份、黄原胶0.35份、纯净水90份。
5.如权利要求3或4所述的植物基酸奶,其特征在于,所述的植物蛋白粉为豆粉、燕麦粉、荞麦粉、青稞粉中的任意一种或两种或多种的组合。
6.如权利要求5所述的植物基酸奶,其特征在于,所述的植物蛋白粉为豆粉。
7.一种植物基酸奶的制备方法,其特征在于,所述的方法包括如下步骤:
(1)按比例称取各原料组分,充分混匀,均质,得到发酵培养基,其中各组分及其重量份数分别为植物蛋白粉3~10份、低聚果糖4~10份、L-赖氨酸0.5~1.5份、乳酸钙0.3~0.6份、牛磺酸0.02~0.04份、黄原胶0.2~0.5份、纯净水80~100份;
(2)将均质后的发酵培养基在115~121℃条件下高温灭菌;
(3)待发酵培养基冷却至37~45℃时,接种活化后的凝结芽孢杆菌VHProbi C08菌液,至发酵培养基中初始活菌量为106~107CFU/mL,37~45℃条件下发酵14~18h;
(4)在4℃条件下酸化12h,即制得植物基酸奶。
8.如权利要求7所述的植物基酸奶,其特征在于,步骤(1)中所述发酵培养液的组分及其重量份数分别为豆粉4份、低聚果糖8份、L-赖氨酸1份、乳酸钙0.4份、牛磺酸0.03份、黄原胶0.35份、纯净水90份。
9.权利要求1-6任一所述植物基酸奶在制备宠物犬猫食品中的应用。
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