CN105325825A - 一种同时提高抗氧化性和去除重金属镉的果蔬汁改良方法 - Google Patents
一种同时提高抗氧化性和去除重金属镉的果蔬汁改良方法 Download PDFInfo
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Abstract
本发明公开了一种同时提高抗氧化性和去除重金属镉的果蔬汁改良方法,属于食品科学技术领域。本发明方法是将植物乳杆菌CCFM8610添加至果蔬汁处理2h或36h后,通过一次简单的离心,即可使果蔬汁中的重金属镉去除率达56.18%至82.87%,并提高了果蔬汁的抗氧化性。本发明方法为食品中重金属的控制和减除提供了新的途径和解决方案。
Description
技术领域
本发明涉及一种同时提高抗氧化性和去除重金属镉的果蔬汁改良方法,属于食品科学技术领域。
背景技术
重金属镉(Cd)是环境和食物链污染中最常见的有毒金属之一,对人体健康都有极其严重的危害。该金属被美国毒性物质与疾病登记署和环境保护署列为第6位危害人类健康的有毒物质,同时被国际癌症研究中心列为一级致癌物。急性镉中毒导致人体出现恶心、呕吐等症状,大剂量摄入甚至会由于急性肝损伤而导致死亡。慢性镉中毒则涉及肝、肾、骨骼、心血管以及生殖器官的损伤。2014年环境保护部和国土资源部发布的全国土壤污染状况调查公报也表明,镉已成为点位超标率最高的污染物(7.0%)。随着镉在工业和生活中的应用,其在水体和土壤中的含量逐年增加,从而进入食物链并最终在人体内蓄积。大量报道表明,镉极易在苹果、草莓、梨、番茄及黄瓜等水果和蔬菜中积累,导致相关果蔬制品镉含量超标,对人体健康构成了威胁。
目前,针对果蔬中镉的去除尚未开发出有效的方法,常见的化学沉降及离子交换等方法存在着安全性低、成本高等缺点,并不适用于果蔬汁镉含量的控制。因此,开发一种简单而有效的去除果蔬汁中镉的方法,是亟待解决的问题。目前也没有涉及利用微生物控制果蔬汁中的有害金属含量的专利或研究。
另外,大量报道表明镉暴露会诱导机体的异常氧化应激反应,导致氧化损伤并进一步造成组织病变,这也是镉对机体的主要致毒机制。若能增强果蔬汁的抗氧化能力,则能使消费者通过摄入果蔬汁而更好地对抗镉毒性。然而发酵菌株、发酵时间均会对抗氧化能力产生影响。
因此,如何对果蔬汁进行改良以同时提高抗氧化性和去除重金属镉,是目前亟需解决的问题。
发明内容
为了克服上述问题,本发明应用植物乳杆菌CCFM8610,开发了简单而有效的去除果蔬汁中镉并提高抗氧化性的方法。
本发明提供了一种果蔬汁高效改良方法,所述改良是指同时提高果蔬汁的抗氧化性、改善果蔬汁风味、降低果蔬汁的镉含量;所述方法是向果蔬汁中添加植物乳杆菌CGMCCNO.6077处理,处理结束后离心去除菌体。
植物乳杆菌CGMCCNO.6077,是发明人在前期工作中从我国传统发酵食品中筛选的到一株植物乳杆菌(Lactobacillusplantarum)CCFM8610,于2012年5月3日在中国微生物菌种保藏管理委员会普通微生物菌种保藏中心保藏,其保藏号为CGMCCNO.6077。。
在本发明的一种实施方式中,所述处理是添加活化后的植物乳杆菌CGMCCNO.6077的菌泥或菌悬液至终浓度为1.2×107至1.5×107cfu/mL。
在本发明的一种实施方式中,所述处理是在37℃下处理2h或36h。
在本发明的一种实施方式中,所述方法具体是:将植物乳杆菌CGMCCNO.6077活化后,离心洗涤,重悬于生理盐水,然后按照终浓度1.2×107至1.5×107的接种量加入果蔬汁中,于37℃下处理2h或36h,然后8000g离心20min,取上清,即为改良后的果蔬汁。
在本发明的一种实施方式中,所述果蔬汁是苹果汁、西红柿汁或者黄瓜汁。
在本发明的一种实施方式中,所述果蔬汁是将水果或蔬菜清洗、切片后按1:2w/v加水,榨汁、离心取上清液、灭菌得到的。
在本发明的一种实施方式中,所述降低果蔬汁的镉含量是指使果蔬汁中的重金属镉去除率达56%以上。
本发明的有益效果:
(1)本发明方法可同时提高果蔬汁的抗氧化性、有效去除重金属镉,并改善果蔬汁风味。按照本发明方法,经短暂处理,即可使果蔬汁中的重金属镉下降67.12%至82.87%;发酵36h后,对自由基的清除率达到54.33%至78.51%,说明果蔬汁的抗氧化能力得到显著提升、本发明方法同时明显改善风味、提高营养水平。
(2)本发明使用的植物乳杆菌是被囊括于卫生部2010年颁布的《可用于食品的菌种名单》的食品级微生物;
具体实施方式
菌株活化和扩培培养基为MRS培养基:MRS培养基是本技术领域的技术人员熟知的培养基,它含有胰蛋白胨、酵母浸膏、葡萄糖、醋酸钠、柠檬酸二铵、吐温80、硫酸镁、硫酸锰,pH6.2~6.4。
果蔬汁的制备:自中国江苏、江西及湖南省三处集市购买新鲜苹果、西红柿及黄瓜(表1)。清洗,切片后按1:2w/v加入超纯水,使用榨汁机压榨以初步获得果蔬汁。将果蔬汁于10000×g离心20分钟后,105℃灭菌10分钟,并分装于玻璃瓶中。
表1本发明中使用果蔬汁的相关信息
实施例1植物乳杆菌CCFM8610处理2h
将植物乳杆菌CCFM8610(即植物乳杆菌CGMCCNO.6077)接种至MRS培养基中活化18h后,离心获得菌体并重悬于灭菌的生理盐水中。按107cfu/mL的添加量将CCFM8610的菌悬液添加入9种不同果蔬汁中,另设不接种乳酸菌的果蔬汁作为空白对照。将果蔬汁在37℃培养2h后,进行标准平板计数。之后以8000×g离心20分钟,取果蔬汁上清。
测定pH值,并加入纯硝酸,于微波消解系统中消解20min,得到的消解液使用原子吸收光谱分析仪测定镉离子含量。将1mL处理前后的果蔬汁与1mL新鲜配制的DPPH无水乙醇溶液(0.2mmol/L)充分混匀后,37℃条件下避光反应30min。将DPPH与PBS(pH7.2)混合作为对照样品,同样条件培养。7000×g离心10min后,在517nm测定吸光度,并按照以下公式计算乳酸菌清除DPPH自由基的能力:
DPPH自由基清除率(%)=[1-A517(样品)/A517(对照)]×100%。
如表2所示,加入植物乳杆菌CCFM8610处理2h后,果蔬汁的pH值出现了明显下降,但果蔬汁中的活菌数并没有出现显著的上升。9类购买于不同地区的果蔬汁中都检测到了重金属镉(表3),表明我国的果蔬产品确实面临着较为严重的镉污染问题。植物乳杆菌CCFM8610处理2h后,果蔬汁中的镉含量出现了显著的下降。西红柿汁3中镉的去除率最低,达到67.12%;黄瓜汁2中镉的去除率最高,达到82.87%。
表2植物乳杆菌CCFM8610处理2h后对果蔬汁中pH和活菌数的影响
注:a,b表示对于pH或活菌数,标有不同字母的组别间存在显著差异(p<0.05)
表3植物乳杆菌CCFM8610处理2h后对果蔬汁中镉含量的影响
注:a,b表示对于镉含量,标有不同字母的组别间存在显著差异(p<0.05)
表4植物乳杆菌CCFM8610处理2h后对果蔬汁抗氧化能力的影响
如表4所示,植物乳杆菌CCFM8610处理2h后,西红柿汁3和西红柿汁3的抗氧化特性得到了显著提升。其中西红柿汁2的DPPH清除率从62.39%上升至69.52%,而西红柿汁3的DPPH清除率从58.83%上升至67.20%。
实施例2植物乳杆菌处理36h
将植物乳杆菌CCFM8610接种至MRS培养基中活化18h后,离心获得菌体并重悬于灭菌的生理盐水中。按107cfu/mL的添加量将CCFM8610的菌悬液添加入9种不同果蔬汁中,另设不接种乳酸菌的果蔬汁作为对照。将果蔬汁在37℃培养36h后,进行标准平板计数。之后以8000×g离心20分钟,取果蔬汁上清,测定pH值,并加入纯硝酸,于微波消解系统中消解20min,得到的消解液使用原子吸收光谱分析仪测定镉离子含量。将1mL处理前后的果蔬汁与1mL新鲜配制的DPPH无水乙醇溶液(0.2mmol/L)充分混匀后,37℃条件下避光反应30min。将DPPH与PBS(pH7.2)混合作为对照样品,同样条件培养。7000×g离心10min后,在517nm测定吸光度,并按照以下公式计算乳酸菌清除DPPH自由基的能力:
DPPH自由基清除率(%)=[1-A517(样品)/A517(对照)]×100%。
如表5所示,加入植物乳杆菌CCFM8610处理36h后,果蔬汁的pH值出现了显著的下降,果蔬汁中活菌数也出现显著上升,每种果蔬汁样本中的活菌数菌突破1×108cfu/mL(除黄瓜汁3中活菌数含量为0.95×108cfu/mL)。经处理36h后,果蔬汁中的镉含量出现了显著的下降(表6),西红柿汁2中镉的去除率最低,达到56.18%;黄瓜汁3中镉的去除率最高,达到81.79%。
表5植物乳杆菌CCFM8610处理36h后对果蔬汁中pH和活菌数的影响
注:a,b表示对于pH或活菌数,标有不同字母的组别间存在显著差异(p<0.05)
表6植物乳杆菌CCFM8610处理36h后对果蔬汁中镉含量的影响
注:a,b表示对于镉含量,标有不同字母的组别间存在显著差异(p<0.05)
表7植物乳杆菌CCFM8610处理36h后对果蔬汁抗氧化能力的影响
如表7所示,植物乳杆菌CCFM8610处理36h后,大多数果蔬汁的抗氧化特性都得到了显著提升。如西红柿汁2的DPPH清除率从62.39%上升至72.69%,而黄瓜汁3的DPPH清除率从54.78%上升至62.75%。
实施例1和2表明,植物乳杆菌CCFM8610可以作为一种食品级的镉吸附剂加入果蔬汁中,仅需2小时的处理时间和一次简单的离心,即可实现对果蔬汁中镉含量的高效去除。同时,若将该菌株作为发酵剂对果蔬汁进行长时间的发酵,同样可以使镉含量去除56%以上。随着发酵的进行,果蔬汁中pH降低,可能会对植物乳杆菌CCFM8610对镉的吸附能力造成负面影响(该菌株在pH为6至7时对镉的吸附能力最强),但发酵过程中细菌数量得到显著升高,在另一方面又可以提升对镉的吸附。这或许是CCFM8610处理36h后同样可以达到2h处理类似的镉去除率的原因。另外CCFM8610的长时间发酵可显著提高果蔬汁的抗氧化能力,这一特性可使处理后的果蔬汁产品对镉毒性造成的氧化损伤具有更好地保护能力。同时,本发明还采用类似的方法,使用植物乳杆菌LP1-7和LP1-2菌株(这两株菌在含有镉的水溶液中同样具有较好的镉吸附效果)对果蔬汁进行改良,处理时间为2h或36h,结果显示,果蔬汁中镉去除率比使用本发明的CCFM8610菌株降低约40%以上,更重要的是,这两株对照菌株基本无法达到较好的抗氧化效果,这可能是由于上述两株菌对果蔬汁的特殊环境适应能力不如CCFM8610菌株。
虽然本发明已以较佳实施例公开如上,但其并非用以限定本发明,任何熟悉此技术的人,在不脱离本发明的精神和范围内,都可做各种的改动与修饰,因此本发明的保护范围应该以权利要求书所界定的为准。
Claims (7)
1.一种果蔬汁高效改良方法,其特征在于,所述方法中的改良是指同时降低果蔬汁的镉含量并提高果蔬汁的抗氧化性;所述方法是向果蔬汁中添加植物乳杆菌CGMCCNO.6077处理,处理结束后离心去除菌体。
2.根据权利要求1所述的方法,其特征在于,所述处理是添加活化后的植物乳杆菌CGMCCNO.6077的菌泥或菌悬液至终浓度为1.2×107至1.5×107cfu/mL。
3.根据权利要求1所述的方法,其特征在于,所述处理是在37℃下处理2h或36h。
4.根据权利要求1所述的方法,其特征在于,所述方法具体是:将植物乳杆菌CGMCCNO.6077活化后,离心洗涤,重悬于生理盐水,然后按照终浓度1.2×107至1.5×107cfu/mL的接种量加入果蔬汁中,于37℃下处理2h或36h,然后8000g离心20min,取上清,即为改良后的果蔬汁。
5.根据权利要求1所述的方法,其特征在于,所述果蔬汁是苹果汁、西红柿汁或者黄瓜汁。
6.根据权利要求1所述的方法,其特征在于,所述降低果蔬汁的镉含量是指使果蔬汁中的重金属镉去除率达56%以上。
7.根据权利要求1所述的方法,其特征在于,所述处理可显著提高果蔬汁对DPPH自由基的清除率,即提升果蔬汁的抗氧化能力。
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