CN106244484A - 南极磷虾共生菌、南极磷虾抗氧化肽及其制备方法和应用 - Google Patents
南极磷虾共生菌、南极磷虾抗氧化肽及其制备方法和应用 Download PDFInfo
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Abstract
本发明涉及一种南极磷虾共生菌、南极磷虾抗氧化肽及其制备方法和应用,属于微生物及生物活性肽技术领域,南极磷虾共生菌嗜冷杆菌NL‑6于2016年6月22日保藏于中国武汉典型培养物保藏中心,保藏号为CCTCCM 2016341。用该菌株发酵南极磷虾匀浆,获得一种南极磷虾抗氧化肽,抗氧化性最高的活性肽组分1~5kDa组分,其羟自由基清除率达到93.0%,还原力为1.32,ABTS清除率为92.4%。所述多肽具有较好的体内外稳定性,更能适应规模化生产的需求,作为抗氧化性食品添加剂具有较好的开发与应用价值。
Description
技术领域
本发明属于微生物及生物活性肽技术领域,具体地说涉及一种南极磷虾共生菌、南极磷虾抗氧化肽及其制备方法和应用。
背景技术
人体内活性氧自由基(ROS)具有一定的功能,如免疫和信号传导过程,在细胞的增殖、分化和凋亡中具有重要作用,但过多的活性氧自由基就会有破坏行为。大量研究已经证明,老化以及老化相关的疾病如癌症、心血管、自身免疫性疾病以及关节炎等的发生机理与体内自由基产生过多或清除自由基能力下降有着密切的关系。因此人们对由自由基引起机体损伤变性,产生的各种疾病引起了强烈的重视。合成的抗氧化剂虽然高效、廉价,但由于有副作用,人们更青睐于选择天然的抗氧化剂。因此低价、高效、低毒的天然抗氧化剂的开发成为研究热点。在医药、化妆品、保健品和食品与饲料添加剂等方面有广阔的应用前景。
微生物发酵技术以现代发酵技术为核心,利用微生物的代谢活动过程,经生物转化而大规模地制造各种工业发酵产品。发酵工程以其生产条件温和,原料来源丰富且价格低廉,产物专一,废弃物对环境的污染小和容易处理的特点在许多领域得到广泛应用。南极磷虾是地球上最大的蛋白质资源,其肌肉蛋白中氨基酸组成全面,配比合理。南极磷虾生物储量丰富,南极磷虾蛋白可作为抗氧化肽的极佳来源。用微生物发酵技术获取南极磷虾抗氧化肽,增加了南极磷虾附加利用价值,提高了南极磷虾资源的高值化和产业化应用前景。2013年,吉林大学的林松毅等酶解蛋清蛋白,并用超滤等技术得到抗氧化性较高的蛋清蛋白源抗氧化肽粉。(公开号:CN 103342734 A);2012年浙江大学的罗自生等以对虾虾壳为原料,用碱性蛋白酶水解,得到抗氧化性较优的小分子抗氧化肽(公开号:CN 102676623 A)。
发明内容
本发明要解决的技术问题在于提供一种南极磷虾共生菌、南极磷虾抗氧化肽及其制备方法和应用,嗜冷杆菌NL-6是从南极磷虾中筛选到的一种南极磷虾共生菌,用该菌株发酵南极磷虾匀浆,获得一种南极磷虾抗氧化肽,其作为食品添加剂具有广阔的应用前景。
一株南极磷虾共生菌嗜冷杆菌NL-6(Psychrobacter sp.NL-6),该嗜冷杆菌NL-6于2016年6月22日保藏于中国典型培养物保藏中心,保藏号为CCTCC M 2016341。
进一步,所述嗜冷杆菌NL-6的16S rDNA长度1439bp,序列见SEQ NO.1。
进一步,所述嗜冷杆菌NL-6为革兰氏阴性菌,菌落特征为白色或乳黄色,表面光滑,有凸起,边缘整齐,扫描电镜图片显示,菌株NL-6呈现短杆棒状,菌体表面光滑,部分刚分裂的菌体呈现椭球状,大部分菌体处于二分裂状态,两个菌体相连,且中间凹陷,连接处细长,菌体间有粘膜产生,单个菌体的大小为:宽:0.6-0.9μm,长:0.8-1.5μm。
本发明还提供一种南极磷虾抗氧化肽,所述南极磷虾抗氧化肽是由嗜冷杆菌NL-6发酵南极磷虾匀浆得到,其步骤如下:
1)菌株培养:
取嗜冷杆菌NL-6菌株,接种于2216E液体培养基中,25-37℃条件,150-220r/min,发酵培养8-14h;
将上述培养后的菌株以体积比4%的接种量接种于含2216E液体培养基的摇瓶中,25-37℃条件,150-220r/min,发酵培养8-14h,即得种子发酵液;
2)南极磷虾冷水解冻,用匀浆机打成匀浆,按质量体积比8-15%湿重量用蒸馏水溶解,121℃高压灭菌20min,制得南极磷虾培养基;
3)将步骤1)制得的种子发酵液以体积比6%-18%接种量接种于步骤2)制备的南极磷虾培养基中,25-37℃,150-220r/min,发酵培养24-48h,即得菌种发酵液;
4)将步骤3)制得的发酵液10000r/min离心30min,收集上清,用分子量为1kDa、5kDa和10kDa的超滤膜超滤,即得<1kDa、1~5kDa、5~10kDa、>10kDa的各组分,将各组分冷冻干燥,即得抗氧化肽粉。
进一步,所述的南极磷虾抗氧化肽优先为1~5kDa。
本发明还提供一种所述的南极磷虾抗氧化肽在食品、食品添加剂及保健品中的应用。
本发明还提供一种所述的1~5kDa南极磷虾抗氧化肽在化妆品中的应用。
本发明与现有技术相比的有益效果:
本发明采用微生物发酵技术获得抗氧化肽,工艺简单,成本低廉,原料丰富易得,反应条件温和,特别是该菌株是从南极磷虾共生菌中筛选获得的一种嗜冷杆菌属的新菌,具有菌源和培养基的同源性,具有较好的发酵效果,避免了采用昂贵的酶试剂,节约了成本。制得的抗氧化肽体外稳定性较好,更能适应规模化生产的需求;且体内稳定性较好,作为抗氧化性食品添加剂具有较好的开发与应用价值。
由于南极磷虾中较高的氟含量是影响其应用的一个主要原因,对本发明南极磷虾抗氧化肽发酵前后冻干粉中氟含量对比发现,发酵后的冻干粉中氟含量(327±3.4mg·kg-1)相比于发酵前(3407±2.5mg·kg-1)降低了10倍之多,且低于欧盟标准(350mg·kg-1),故发酵后的南极磷虾抗氧化肽直接作为蛋白粉,或作为食品、保健品的添加剂都具有较好的应用前景。
南极磷虾抗氧化肽各组分的抗氧化性在体外有较好的稳定性,在高温(100℃煮沸1h)条件下,各组分羟自由基清除活性均可保持在98%以上,还原能力均可保持在84%以上,其中1~5kDa和5~10kDa组分在各pH(2、4、6、8和10)下还原能力均可保持在96%以上;故由<5kDa的抗氧化肽作为化妆品添加剂,具有稳定性好、分子量小、易于吸收的优势。
南极磷虾抗氧化肽各组分对有Ca2+存在的条件下,羟自由基清除率、还原能力均能保持在90%以上,故由<5kDa的抗氧化肽作为食品添加剂可与各类钙类产品同服,其抗氧化性能几乎不受影响。
南极磷虾抗氧化肽各组分抗氧化性在以Pepsase-Trypsin-Chymotrypsin复合酶体系模拟的体内胃肠道环境中有较好的稳定性,1~5kDa组分羟自由基清除活性经胃环境模拟后活性保持在98.6%,经胃肠环境模拟后活性保持在84.7%;还原能力经胃环境模拟后活性保持在96.7%,经胃肠环境模拟后活性保持在82.0%,更进一步验证了1~5kDa小分子量抗氧化肽具有较好的体内外稳定性,作为食品添加剂的有较好的应用价值。
附图说明
图1嗜冷杆菌NL-6的扫描电镜图;
图2嗜冷杆菌NL-6的系统发育树;
嗜冷杆菌NL-6,拉丁文为Psychrobacter sp.NL-6,于2016年6月22日保藏于中国典型培养物保藏中心,保藏号为CCTCC M 2016341,保藏地址为中国武汉武汉大学。
具体实施方式
下面通过实施例来进一步说明本发明的技术方案,但本发明的保护范围不受实施例任何形式的限制。
实施例1菌株NL-6的筛选
1、具抗氧化活性发酵液的获得
1)共生微生物菌株的筛选将南极磷虾洗净,研钵研磨,取1g研磨匀浆于无菌蒸馏水中,150-220r/min,振荡1~2h,取混悬液于蒸馏水中,梯度稀释,涂布于2216E固体培养基上,25-37℃培养24-48h,挑取外观形态不同的10株菌(编号NL-(1~10))于2216E液体培养基中培养,保存菌种。
2)菌株活化培养:
将10株共生菌株接种于2216E液体培养基的试管中,25-37℃条件,150-220r/min,发酵培养8-14h。
将上述培养后的菌株以体积比4%的接种量接种于含2216E液体培养基的摇瓶中,25-37℃条件,150-220r/min,发酵培养8-14h,即得种子发酵液。
3)南极磷虾冷水解冻,用匀浆机打成匀浆,按质量体积比8-15%(湿重量)用蒸馏水溶解,121℃高压灭菌20min,制得南极磷虾培养基。
4)将制得的种子发酵液以体积比6%-18%接种量接种于南极磷虾培养基中,25-37℃,150-220r/min,发酵培养24-48h,即得菌种发酵液。
5)将菌种发酵液10000r/min离心30min,取上清液。
2、抗氧化性测定
1)羟自由基清除率测定
在10mL试管中加入2mL的待测液,然后依次加入2mL浓度为9mmol/L的FeSO4,2mL浓度为9mmol/L的水杨酸,最后加入2mL浓度为9mmol/L的H2O2启动反应,静置10min后于510nm处测定吸光度A1。考虑到提取液本身的吸光值,做试样空白,测出扣除试样空白的吸光度A2,同时测定空白对照液的吸光度A0。
清除率(%)=(A0-A1+A2)/A0×100
2)还原力测定
加入步骤1制备的各上清液2.5mL,同时加入2.5mL的磷酸缓冲液(0.2mol/L,pH6.6)和2.5mL 1%的铁氰化钾溶液。混合物50℃水浴20min后加入2.5mL 10%的三氯乙酸,室温静置10min。取2.5mL反应液,加入2.5mL蒸馏水和0.5mL0.1%氯化铁溶液。反应10min后测定700nm处吸光值,该值越高说明样品的还原性越强。
表1 10株共生微生物发酵液抗氧化性大小比较
菌株 | 羟自由基清除率 | 还原力 |
NL-1 | 89.8% | 0.487 |
NL-2 | 78.4% | 0.270 |
NL-3 | 79.5% | 0.388 |
NL-4 | 85.4% | 0.441 |
NL-5 | 87.8% | 0.453 |
NL-6 | 91.7% | 0.604 |
NL-7 | 74.3% | 0.414 |
NL-8 | 68.3% | 0.484 |
NL-9 | 75.9% | 0.440 |
NL-10 | 81.4% | 0.375 |
通过表1我们选取了羟自由基清除率和还原力均较好的菌株NL-6进行进一步研究,并对菌株NL-6进行了菌种鉴定。
实施例2
菌株NL-6的菌种鉴定
(1)形态特征
革兰氏染色结果显示,菌株NL-6为革兰氏阴性菌,菌落特征为白色或乳黄色,表面光滑,有凸起,边缘整齐。扫描电镜图片显示,菌株NL-6呈现短杆棒状,菌体表面光滑,部分刚分裂的菌体呈现椭球状,大部分菌体处于二分裂状态,两个菌体相连,且中间凹陷,连接处细长,菌体间有粘膜产生,见图1。由图1可以大体推算单个菌体的大小为:宽:0.6-0.9μm,长:0.8-1.5μm。
(2)生理生化反应特征
由于革兰氏染色显示,该菌为革兰氏阴性菌,故可以用API 20E细菌鉴定系统进行生理生化分析。结果见表2
表2菌株NL-6的生理生化鉴定结果
注:“-”表阴性;“+”表示阳性
由该表可以知道,菌株NL-6可以发酵利用葡萄糖、鼠李糖、密二糖、阿拉伯糖。
(3)16S rDNA序列分析
吸取1.5mL的处于对数生长期的菌株NL-6菌液,4℃,10000r/min,离心10min。弃上清液,加入400μL的TE缓冲液,用枪头将沉淀吹打均匀,加入10μL的溶菌酶,混合均匀,置于37℃水浴中90min,4℃,10000r/min,离心10min,保留上清即为DNA溶液。
采用16S rDNA通用引物,由青岛华大基因合成。正向引物:27F 5’-AGAGTTTGATCCTGGTCAG-3’,反向引物:1492R 5’-CGGCTACCTTGTTACGACTT-3’;16S rDNA序列的PCR条件是:95℃预变性5min;95℃变性30s,55℃退火30s,72℃延伸1min,循环30次,72℃延伸10min。PCR反应体系是:反应体系:50μL,模板DNA:2μL,上游引物:2μL,下游引物:2μL,2X Easy tap MasterMix:25μL,双蒸水:19μL。
用1%的琼脂糖电泳检测PCR扩增产物,以D2000Marker为标准分子量对照,约在1500bp处有一条PCR扩增产物条带,对菌株NL-6的16S rDNA PCR产物送到青岛华大基因进行测序。序列见SEQ NO.1,测得菌株NL-6的16S rDNA序列长度为1439bp。
将菌株NL-6的16S rDNA序列在NCBI的核酸数据库中通过BLAST进行分析。用BioEdit软件进行多序列对比,与菌株NL-6的16S rDNA序列(1439bp)相似性比较高的菌株为Psychrobacter fozii NF23(T)(98.82%)、Psychrobacter okhotskensis MD17(T)(98.47%)、Psychrobacter cryohalolentis K5(T)(98.33%)。下载与实验菌株亲缘关系较近的序列,通过MEGA6软件Neighbor-joining选项绘制发育树如图2。
通过《常见细菌系统鉴定手册》,可以确定该菌株属于嗜冷杆菌属(Psychrobacter)的细菌,命名为Psychrobacter sp.NL-6,可能为嗜冷杆菌属的新种。
实施例3抗氧化肽的制备
1)菌株的活化
将南极磷虾共生菌嗜冷杆菌NL-6接种于2216E液体培养基的试管中,28℃,200r/min,培养10h;将上述培养后的菌株以4%接种量(体积比)接种于含发酵培养基的摇瓶中,28℃条件,200r/min,发酵培养10h。即得种子发酵液。
2)NL-6发酵南极磷虾
南极磷虾冷水解冻,用匀浆机打成匀浆,按质量体积比10%(湿重量)用蒸馏水溶解,121℃高压灭菌20min,制得南极磷虾培养基。
将制得的种子发酵液以体积比8%接种量接种于南极磷虾培养基中,32℃,200r/min,发酵培养48h,即得菌种发酵混悬液。
3)各组分抗氧化肽制备
将制得的发酵液10000r/min离心30min,收集上清,用分子量为1kDa、
5kDa、10kDa的超滤膜超滤,即得<1kd、1~5kDa、5~10kDa、>10kDa的各
组分,将各组分冷冻干燥,即得抗氧化肽粉。
经验证,本发明所得的抗氧化性最高的活性肽组分为1~5kDa组分,其
羟自由基清除率达到93.0%,还原力为1.32,ABTS清除率为92.4%。
实施例4体外抗氧化性稳定性测定
温度条件测定
将实施例3获得的各组分分别在20℃、40℃、60℃、80℃、100℃条件下恒温水浴1h,放置室温后测其羟自由基清除率,还原力大小。实验证实,各组分羟自由基清除活性保持在98%以上,还原能力保持在84%以上。
pH条件测定
将实施例3获得的各组分分别溶解于pH为2、4、6、8和10的缓冲液中,测其还原力大小。实验证实,1~5kDa和5~10kDa组分在各pH下还原能力均可保持在96%以上;
Ca2+影响
将实施例3获得的各组分溶解于浓度为5mmol/L的Ca2+溶液中,测其抗氧化活性影响。实验证实,各组分对有Ca2+存在的条件下,羟自由基清除率、还原能力均能保持在90%以上,故由<5kDa的抗氧化肽制得的食品添加剂,分子量小,易于吸收,且可与各类钙类产品同服,其抗氧化性能几乎不受影响。
实施例5体内抗氧化性稳定性测定
采用Pepsase-Trypsin-Chymotrypsin复合酶体系,模拟胃肠道环境对南极磷虾抗氧化肽活性影响。
1)胃蛋白酶酶液的制备
配置pH为2.0的氯化钾-盐酸缓冲液,待用。准确称取1.00g Pepsase溶于100mL已配置的人工胃液,现配现用。
2)人工肠液的制备取50mL胃蛋白酶酶解液,配置2.0M的NaOH溶液调节其pH为7.0,再加入1%(w/w)的Trypsin和Chymotrypsin(1:1)。
3)模拟胃中的消化环境用分析天平称取1.00g南极磷虾多肽冻干粉(P5组分),溶于50mL的胃蛋白酶酶解液(37±1)℃,100r/min条件下振荡2h,将其取出,于沸水浴中煮沸10min,使胃蛋白酶失活,终止反应,冷却至室温,并用2M NaOH调节pH值至7.0。取25mL消化液于12000r/min的离心机离心30min,收集清液,测定其抗氧化活性。
4)模拟肠道中的消化环境取出已在胃蛋白酶酶解液中消化2h的样品25mL,调节PH为7.0,加入人工肠液25mL,于(37±1)℃,100r/min条件下振荡2h,将其取出,于沸水浴中煮沸10min,使其含有的蛋白酶失活,终止反应,冷却至室温,离心,调节转速为12000r/min,离心30min,并检测上清液的自由基清除率。
经证实,各组分抗氧化性在以Pepsase-Trypsin-Chymotrypsin复合酶体系模拟的胃肠道环境中有较好的稳定性,1~5kDa组分羟自由基清除活性经胃环境模拟后活性保持在98.6%,经胃肠环境模拟后活性保持在84.7%;还原能力经胃环境模拟后活性保持在96.7%,经胃肠环境模拟后活性保持在82.0%。
Claims (9)
1.一株南极磷虾共生菌嗜冷杆菌NL-6,其特征在于该嗜冷杆菌NL-6于2016年6月22日保藏于中国典型培养物保藏中心,保藏号为CCTCC M 2016341。
2.根据权利要求1所述的一株南极磷虾共生菌嗜冷杆菌NL-6,其特征在于所述嗜冷杆菌NL-6的16S rDNA长度1439bp,序列见SEQ NO.1。
3.根据权利要求1所述的一株南极磷虾共生菌嗜冷杆菌NL-6,其特征在于所述嗜冷杆菌NL-6为革兰氏阴性菌,菌落特征为白色或乳黄色,表面光滑,有凸起,边缘整齐,扫描电镜图片显示,菌株NL-6呈现短杆棒状,菌体表面光滑,部分刚分裂的菌体呈现椭球状,大部分菌体处于二分裂状态,两个菌体相连,且中间凹陷,连接处细长,菌体间有粘膜产生,单个菌体的大小为:宽:0.6-0.9μm,长:0.8-1.5μm。
4.一种南极磷虾抗氧化肽,其特征在于所述南极磷虾抗氧化肽是由权利要求1-3任何一项所述的嗜冷杆菌NL-6发酵南极磷虾匀浆得到,其步骤如下:
1)菌株培养:
取嗜冷杆菌NL-6菌株,接种于2216E液体培养基中,25-37℃条件,150-220r/min,发酵培养8-14h;
将上述培养后的菌株以体积比4%的接种量接种于含2216E液体培养基的摇瓶中,25-37℃条件,150-220r/min,发酵培养8-14h,即得种子发酵液;
2)南极磷虾冷水解冻,用匀浆机打成匀浆,按质量体积比8-15%湿重量用蒸馏水溶解,121℃高压灭菌20min,制得南极磷虾培养基;
3)将步骤1)制得的种子发酵液以体积比6%-18%接种量接种于步骤2)制备的南极磷虾培养基中,25-37℃,150-220r/min,发酵培养24-48h,即得菌种发酵液;
4)将步骤3)制得的发酵液10000r/min离心30min,收集上清,用分子量为1kDa、5kDa和10kDa的超滤膜超滤,即得<1kDa、1~5kDa、5~10kDa、>10kDa的各组分,将各组分冷冻干燥,即得南极磷虾抗氧化肽粉。
5.根据权利要求4所述的一种南极磷虾抗氧化肽,其特征在于所述的南极磷虾抗氧化肽为1~5kDa。
6.一种权利要求4所述的南极磷虾抗氧化肽在食品、食品添加剂及保健品中的应用。
7.含有权利要求4所述的南极磷虾抗氧化肽的食品或保健品。
8.一种权利要求5所述的南极磷虾抗氧化肽在化妆品中的应用。
9.含有权利要求5所述的南极磷虾抗氧化肽的化妆品。
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---|---|---|---|---|
CN107841519A (zh) * | 2017-12-06 | 2018-03-27 | 华南协同创新研究院 | 一种南极磷虾抗氧化肽及其制备方法与应用 |
CN108130352A (zh) * | 2017-12-06 | 2018-06-08 | 华南协同创新研究院 | 一种制备南极磷虾抗氧化肽的方法及该抗氧化肽与应用 |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102559825A (zh) * | 2012-01-18 | 2012-07-11 | 辽宁省大连海洋渔业集团公司 | 一种制备南极磷虾低氟水解多肽的方法 |
CN105506039A (zh) * | 2016-02-01 | 2016-04-20 | 浙江工业大学 | 米曲霉发酵虾壳制备抗氧化肽的方法 |
-
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Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102559825A (zh) * | 2012-01-18 | 2012-07-11 | 辽宁省大连海洋渔业集团公司 | 一种制备南极磷虾低氟水解多肽的方法 |
CN105506039A (zh) * | 2016-02-01 | 2016-04-20 | 浙江工业大学 | 米曲霉发酵虾壳制备抗氧化肽的方法 |
Non-Patent Citations (2)
Title |
---|
JIANAN SUN ET AL.: "Screening of Microorganisms from Deep-Sea Mud for Antarctic Krill (Euphausia superba) Fermentation and Evaluation of the Bioactive Compounds", 《APPL BIOCHEM BIOTECHNOL》 * |
江艳华 等: "基于高通量测序的冷冻南极磷虾中细菌菌群结构分析", 《食品安全质量检测学报》 * |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN107841519A (zh) * | 2017-12-06 | 2018-03-27 | 华南协同创新研究院 | 一种南极磷虾抗氧化肽及其制备方法与应用 |
CN108130352A (zh) * | 2017-12-06 | 2018-06-08 | 华南协同创新研究院 | 一种制备南极磷虾抗氧化肽的方法及该抗氧化肽与应用 |
CN108130352B (zh) * | 2017-12-06 | 2020-08-25 | 华南协同创新研究院 | 一种制备南极磷虾抗氧化肽的方法及该抗氧化肽与应用 |
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