CN116076692A - 一种富含功能食品因子调味剂中添加剂及其制备方法 - Google Patents
一种富含功能食品因子调味剂中添加剂及其制备方法 Download PDFInfo
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Abstract
本发明属于水产品生物加工技术领域,具体涉及一种富含功能食品因子调味剂中添加剂及其制备方法。将原料粉碎后加入清水煮沸,静止放置冷却至30℃‑60℃,获得原料原液;向原料原液中加入蛋白酶进行酶解,得到原料酶解液;调节pH值,高温灭菌得到原料培养基;向冷却至室温的原料培养基中接种乳酸菌并添加反应底物,28℃‑45℃条件下培养1‑5天获得富含功能食品因子的发酵液,即为添加剂。本发明的方法能够高值化利用扇贝加工废弃物,制得的产品总氨基酸含量、风味氨基酸含量均显著提升,并富含功能食品因子,具有改善机体睡眠质量、增强记忆力、抗氧化、抗衰老、降血压等生理功效。
Description
技术领域
本发明属于水产品生物加工技术领域,具体涉及一种富含功能食品因子调味剂中添加剂及其制备方法。
背景技术
我国是扇贝养殖大国,资源极为丰富,从北到南的整个沿海地区都大量盛产。目前对于贝类的整体利用率较低,仅以鲜食、冷冻品及传统干腌制品为主。在扇贝加工过程中会产生的大量(20%-30%)未被充分利用的副产物裙边、内脏等,含有丰富的氨基酸、多糖、蛋白质、微量元素和无机物质,具有较高的营养价值和丰富的生理活性成分,是开发制备高附加值保健食品及功能性食品的理想原料。目前,市场上的扇贝裙边制品仅有裙边小食品、裙边罐头等,只是简单地加工利用了其风味,未对其活性成分进行高值利用,既浪费资源又污染环境,成为我国水产业亟待解决的大问题。开展扇贝资源高值利用研究,提取、优化和改造它们所含独特功能物质开发成为新型生物制品,贝类资源可持续利用的重要方向。采用生物酶解技术、微生物发酵技术及膜分离技术,可制备出富含牛磺酸、多肽及氨基酸的海鲜调味品和纯天然保健产品。
随着生活水平提高,人们对调味料的需求已由单一的鲜味型向鲜味、营养、方便的复合型转变。
发明内容
本发明的目的在于提供一种富含功能食品因子调味剂的制备方法。
为实现上述目的,本发明采用的技术方案为:
一种富含功能食品因子调味剂中添加剂的制备方法:
1)将原料粉碎后加入其质量1-10倍体积的清水(W/V)煮沸,静止放置冷却至30℃-60℃,获得原料原液;
2)向原料原液中加入质量分数为0.5%-8.0%(W/V)的蛋白酶进行酶解,酶解温度28℃-65℃,酶解时间2-24小时,得到原料酶解液;
3)对原料酶解液调节pH值3.0-7.0,高温灭菌得到原料培养基;
4)向冷却至室温的原料培养基中接种0.5%-4%(V/V)活化好的乳酸菌,在28-45℃条件下培养8-48小时,获得富含乳酸菌的原料发酵液;
5)向富含乳酸菌的原料发酵液当中添加质量分数0.1%-5%(W/V)反应底物,28℃-45℃条件下培养1-5天获得富含功能食品因子的发酵液,即为添加剂。
所述步骤3)向原料酶解液中添加0.5%-3.0%(W/V)的碳源,调节pH值3.0-7.0,高温灭菌得到原料培养基。
所述原料为扇贝加工废弃物——扇贝外套膜和内脏其中的一种或两种混合。
所述蛋白酶可以为胰蛋白酶,木瓜蛋白酶,酸性蛋白酶,碱性蛋白酶,风味蛋白酶,复合蛋白酶,胃蛋白酶,中性蛋白酶,专动物蛋白复合蛋白酶中的一种或几种的混合。
所述碳源为葡萄糖、乳糖、蔗糖中的一种或几种的混合。
所述乳酸菌为瑞士乳杆菌、乳双歧杆菌、两双歧杆菌、保加利亚乳杆菌动物双歧杆菌、短乳杆菌中的一种或几种的混合。
所述反应底物为谷氨酸钠。
一种所述方法制备所得富含功能食品因子调味剂中添加剂,其特征在于:所述添加剂为按权利要求1所述方法制备获得富含高浓度γ-氨基丁酸(GABA)的调味剂中添加剂。
一种富含功能食品因子调味剂,调味剂中添加所述的添加剂。
所述调味剂成分按照如下比例进行配比:所述的添加剂45%-60%(V/V)、酱油20%-25%(V/V)、食盐5%-8%(W/V)、葡萄糖2%-10%(W/V)、食用淀粉0.5%-3.5%(W/V)、余量为水,混合加热(加热煮沸5-30分钟至淀粉糊化,而后冷却至室温)后即得到富含功能食品因子调味剂。
所述添加剂为权利要求1所述富含功能食品因子的发酵液经浓缩至原体积的1-1/5(V/V),待用。
乳酸菌的活化参见《喷雾干燥型乳酸菌制剂活化增殖条件的研究》(张大凤等,2012)。GABA的检测方法参见《GABA含量检测方法Q/LJS0006S-2013》。
本发明所具有的优点
1.本发明所使用的技术使扇贝加工下脚料发酵液中总氨基酸含量提升80%,风味氨基酸含量提升50%,使调味料成品的风味更佳。
2.本发明所获得的发酵液中功能食品因子GABA的含量丰富,可达8g/L,利用1吨扇贝加工废弃物可生产50千克GABA,极大提升了GABA的产量和扇贝加工废弃物的利用率。
3.本发明所获得的调味剂利用葡萄糖、乳糖、蔗糖等碳源和一定量的食盐形成高渗透压,降低产品的水分活度,并通过乳酸菌所产生的天然乳酸降低产品的pH值。通过水份活度和pH值的协同效果,抑制微生物的生长。本发明制得的产品,保留了扇贝特有的鲜味和滋味,和常规的产品比较,含盐量低且不需要添加防腐剂,并富含功能食品因子GABA,具有改善机体睡眠质量、增强记忆力、抗氧化、抗衰老、降血压等生理功效。和常规的产品比较,营养价值高、风味更加、含盐量低且不需要添加防腐剂,是新一代健康营养型调味品。
具体实施方式
下面对本发明作进一步说明,并且本发明的保护范围不仅局限于以下实施例。
本发明中利用扇贝加工废弃物获得富含功能食品因子(GABA)的调味剂,所得的产品总氨基酸含量、风味氨基酸含量均显著提升,并富含功能食品因子,具有改善机体睡眠质量、增强记忆力、抗氧化、抗衰老、降血压等生理功效。和常规的产品比较,营养价值高、风味更加、含盐量低且不需要添加防腐剂,是新一代健康营养型调味品。
实施例1
1)将风干的扇贝外套膜粉碎后加入其质量10倍体积的清水(W/V)煮沸,静止放置冷却至30℃,获得原料原液;
2)向原料原液中加入其体积质量分数为1%(W/V)的专动物蛋白复合蛋白酶(市购)进行酶解,酶解温度50℃,酶解时间8小时,得到原料酶解液;
3)原料酶解液调节pH值4.5,高温灭菌得到原料培养基;
4)向冷却至室温的原料培养基中接种2%(V/V)活化好的短乳杆菌,在30℃条件下培养24小时,获得富含短乳杆菌的原料发酵液;
5)向富含短乳杆菌的原料发酵液当中添加其体积质量分数1%(W/V)的反应底物,37℃条件下培养4天获得富含功能食品因子(GABA)的发酵液,经检测发酵液中GABA含量为3.42g/L;其中,反应底物为谷氨酸钠。
6)将富含功能食品因子的发酵液浓缩至原体积的1/2(V/V),获得浓缩汁,
将得浓缩液与其他成分按照如下比例进行配比:浓缩汁50%(V/V)、酱油20%(V/V)食盐5%(W/V)、葡萄糖2%(W/V)、食用淀粉2%(W/V)、余量为水,混合加热(加热煮沸5-30分钟至淀粉糊化,而后冷却至室温)后即得到富含功能食品因子调味剂,GABA含量为310mg/100g。
*乳酸菌的活化参见《喷雾干燥型乳酸菌制剂活化增殖条件的研究》(张大凤等,2012)。GABA的检测方法参见《GABA含量检测方法Q/LJS0006S-2013》。短乳杆菌购买于北京百欧博伟生物技术有限公司编号为bio-53246。
实施例2
1)将风干的扇贝外套膜粉碎后加入其质量5倍体积的清水(W/V)煮沸,静止放置冷却至30℃,获得原料原液;
2)准备9份相同的原料原液,每一份原料原液中加入不同的蛋白酶(胰蛋白酶、木瓜蛋白酶、酸性蛋白酶、碱性蛋白酶、风味蛋白酶、复合蛋白酶、胃蛋白酶、中性蛋白酶、专动物蛋白复合蛋白酶)进行酶解,蛋白酶质量分数为1%(W/V),以酶的最适反应温度和pH值进行酶解,酶解时间8小时,得到原料酶解液,测定酶解液中17种氨基酸及风味氨基酸含量(见表1和表2);
3)每一份原料酶解液再添加0.5%(W/V)的葡萄糖作为碳源,调节pH值4.5,高温灭菌得到原料培养基;
4)向冷却至室温的原料培养基中接种1%(V/V)活化好的短乳杆菌,在37℃条件下培养21小时,获得富含短乳杆菌的原料发酵液;
5)向富含短乳杆菌的原料发酵液当中添加质量分数1%(W/V)的谷氨酸钠,37℃条件下培养5天,检测发酵液中GABA含量(表3);
*乳酸菌的活化参见《喷雾干燥型乳酸菌制剂活化增殖条件的研究》(张大凤等,2012)。GABA的检测方法参见《GABA含量检测方法Q/LJS0006S-2013》。发酵液中氨基酸的检测方法参见《高效液相色谱法测定谷氨酸棒杆菌发酵液中17种氨基酸》(王平等,2017)。短乳杆菌购买于北京百欧博伟生物技术有限公司编号为bio-53246。
表1 不同蛋白酶解液中17种氨基酸含量(%)
表2 不同蛋白酶酶解液风味氨基酸含量(%)
表3 不同蛋白酶酶解液经短乳杆菌发酵后GABA含量(g/L)
由表1可知,专动物蛋白复合蛋白酶酶解使扇贝加工下脚料酶解液中总氨基酸含量提升80%(与酸性蛋白酶酶解相比);由表2可知,专动物蛋白复合蛋白酶酶解使扇贝加工下脚料酶解液中风味氨基酸含量提升50%(与酸性蛋白酶酶解相比);由表3可知,专动物蛋白复合蛋白酶酶解使扇贝加工下脚料发酵液中GABA的含量提升11倍(与碱性蛋白酶相比)。
实施例3
1)将风干的扇贝外套膜粉碎后加入其质量5倍体积的清水(W/V)煮沸,静止放置冷却至30℃,获得原料原液;
2)准备23份相同的原料原液,每一份原料原液中加入专动物蛋白复合蛋白酶进行酶解,蛋白酶质量分数为1%(W/V),55℃酶解时间8小时,得到原料酶解液;
3)调节酶解液pH值至4.5,高温灭菌得到原料培养基;
4)分别向冷却至室温的23份原料培养基中分别接种1%(V/V)活化好的23种不同乳酸菌,在37℃条件下培养21小时,获得富含乳酸菌的原料发酵液;
5)向富含乳酸菌的原料发酵液当中添加质量分数1%(W/V)的谷氨酸钠,37℃条件下培养4天,检测发酵液中GABA含量(见表4);
*乳酸菌的活化参见《喷雾干燥型乳酸菌制剂活化增殖条件的研究》(张大凤等,2012)。GABA的检测方法参见《GABA含量检测方法Q/LJS0006S-2013》。短乳杆菌购买于北京百欧博伟生物技术有限公司编号为bio-53246,其他22种乳酸菌购买于镇江市天益生物科技有限公司。
表4 不同乳酸菌产GABA(g/L)能力
由表4可知,不同乳酸菌利用扇贝下脚料生产GABA的能力不同,其中短乳杆菌产GABA能力最强,可达6.180g/L。瑞士乳杆菌、乳双歧杆菌、两双歧杆菌、保加利亚乳杆菌动物双歧杆菌发酵液种GABA含量分别为0.237g/L、0.228g/L、0.227g/L、0.219g/L和0.233g/L。
实施例4
1)将风干的扇贝外套膜和扇贝内脏混合物粉碎后加入其质量10倍体积的清水(W/V)煮沸,静止放置冷却至30℃,获得原料原液;
2)向原料原液中加入专动物蛋白复合蛋白酶进行酶解,蛋白酶质量分数为1%(W/V),50℃酶解时间10小时,得到原料酶解液;
3)调节酶解液pH值至5.0,高温灭菌得到原料培养基;
4)向冷却至室温的原料培养基中接种2%(V/V)活化好的短乳杆菌,在37℃条件下培养24小时,获得富含乳酸菌的原料发酵液;
5)向富含乳酸菌的原料发酵液当中添加质量分数1.5%(W/V)的谷氨酸钠,37℃条件下培养5天,检测发酵液中GABA含量为5.42g/L,1吨扇贝加工废弃物(风干扇贝外套膜和内脏)可制备50千克以上GABA。
*乳酸菌的活化参见《喷雾干燥型乳酸菌制剂活化增殖条件的研究》(张大凤等,2012)。GABA的检测方法参见《GABA含量检测方法Q/LJS0006S-2013》。短乳杆菌购买于北京百欧博伟生物技术有限公司编号为bio-53246。
对比例1
将实施例2中采用专动物蛋白复合蛋白酶并在碳源的添加下制备所得富含功能食品因子的发酵液浓缩(①),分别与不同方法制备所得富含GABA的调味品进行比较。
不同方法制备所得富含GABA的调味品分别为
②申请号为CN201510335574.9,发明名称为《一种富含γ-氨基丁酸的海鲜调味料及其制备方法》制备所得液体调味料中实施例3;
③申请号为CN201510335574.9,发明名称为《一种富含γ-氨基丁酸的海鲜调味料及其制备方法》固体调味料中实施例1。
将上述不同方法获得富含GABA的调味品进行检测,GABA的检测方法参见《GABA含量检测方法Q/LJS0006S-2013》(参见表5)。
表5 不同方法制备调味剂GABA(g/L)含量比较
① | ② | ③ |
8.01g/kg | 100mg/kg | 1g/kg |
*①实施例2制备的调味料中GABA的含量。
由表5可知,本发明所获得的调味料中GABA的含量远高于其他方法制备所得调味品中GABA的含量,其是通过菌种的筛选和培养策略的改变所实现的。首先,不同菌种生产GABA的能力有差异,因此需要对发酵菌种进行优选。其次,利用微生物产GABA关键是谷氨酸脱羧酶(GAD)的产生。耐酸系统主要通过GAD和相关的Glu(谷氨酸)/GABA反向转运子协同发挥作用。反向转运子将外源谷氨酸转运到细胞内,在GAD作用下脱羧,不可逆地消耗了H+,从而维持细胞的生理pH。GAD基因只有被酸性、高渗、低渗和低氧环境诱导,并在H-NS和RpoS因子的分子级联调控下才能获得表达。在扇贝溶液中加入适量的葡萄糖,一可以弥补扇贝缺失的糖分,为微生物生长提供足够的碳原,促进微生物的生长繁殖,发酵产酸;二可以稍提高细胞外渗透压,有利于诱导GAD基因表达。通常微生物来源的GAD最适反应pH值在3.8~5.0,主要集中在pH值4.0~5.0,当pH值高于6.0时易发生离解。因此,适当降低培养基初始pH值不仅可以诱导GAD基因表达,提高GAD活性与稳定性,还可以补充GABA产生所消耗掉的H+,有利于发酵液中GABA的富集。
Claims (10)
1.一种富含功能食品因子调味剂中添加剂的制备方法,其特征在于:
1)将原料粉碎后加入其质量1-10倍体积的清水(W/V)煮沸,静止放置冷却至30℃-60℃,获得原料原液;
2)向原料原液中加入质量分数为0.5%-8.0%(W/V)的蛋白酶进行酶解,酶解温度28℃-65℃,酶解时间2-24小时,得到原料酶解液;
3)对原料酶解液调节pH值3.0-7.0,高温灭菌得到原料培养基;
4)向冷却至室温的原料培养基中接种0.5%-4%(V/V)活化好的乳酸菌,在28-45℃条件下培养8-48小时,获得富含乳酸菌的原料发酵液;
5)向富含乳酸菌的原料发酵液当中添加质量分数0.1%-5%(W/V)反应底物,28℃-45℃条件下培养1-5天获得富含功能食品因子的发酵液,即为添加剂。
2.按权利要求1所述的富含功能食品因子调味剂中添加剂的制备方法,其特征在于:所述步骤3)向原料酶解液中添加0.5%-3.0%(W/V)的碳源,调节pH值3.0-7.0,高温灭菌得到原料培养基。
3.按权利要求1所述的富含功能食品因子调味剂中添加剂的制备方法,其特征在于:所述原料为扇贝加工废弃物——扇贝外套膜和内脏其中的一种或两种混合。
4.按权利要求1所述的富含功能食品因子调味剂中添加剂的制备方法,其特征在于:所述蛋白酶可以为胰蛋白酶,木瓜蛋白酶,酸性蛋白酶,碱性蛋白酶,风味蛋白酶,复合蛋白酶,胃蛋白酶,中性蛋白酶,专动物蛋白复合蛋白酶中的一种或几种的混合。
5.按权利要求2所述的富含功能食品因子调味剂中添加剂的制备方法,其特征在于:所述碳源为葡萄糖、乳糖、蔗糖中的一种或几种的混合。
6.按权利要求1所述的富含功能食品因子调味剂中添加剂的制备方法,其特征在于:所述乳酸菌为瑞士乳杆菌、乳双歧杆菌、两双歧杆菌、保加利亚乳杆菌动物双歧杆菌、短乳杆菌中的一种或几种的混合。
7.按权利要求1所述的富含功能食品因子调味剂中添加剂的制备方法,其特征在于:所述反应底物为谷氨酸钠。
8.一种权利要求1所述方法制备所得富含功能食品因子调味剂中添加剂,其特征在于:所述添加剂为按权利要求1所述方法制备获得富含高浓度γ-氨基丁酸(GABA)的调味剂中添加剂。
9.一种富含功能食品因子调味剂,其特征在于:调味剂中添加权利要求1所述的添加剂。
10.按权利要求9所述的富含功能食品因子调味剂,其特征在于:所述调味剂成分按照如下比例进行配比:权利要求1所述的添加剂45%-60%(V/V)、酱油20%-25%(V/V)、食盐5%-8%(W/V)、葡萄糖2%-10%(W/V)、食用淀粉0.5%-3.5%(W/V)、余量为水,混合加热后即得到富含功能食品因子调味剂。
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