CN107814832B - 一种马鲛鱼蛋白来源的铁螯合肽及其应用 - Google Patents
一种马鲛鱼蛋白来源的铁螯合肽及其应用 Download PDFInfo
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Abstract
本发明涉及生物技术领域，尤其涉及一种马鲛鱼蛋白来源的铁螯合肽及其应用，所述马鲛鱼蛋白来源的铁螯合肽的氨基酸序列为Gln‑Lys‑Gly‑Thr‑Tyr‑Asp‑Asp‑Tyr‑Val‑Glu‑Gly‑Leu。本发明马鲛鱼蛋白来源的铁螯合肽具有很强的铁螯合能力，可应用于铁补充剂，安全无毒副作用、促进铁吸收，既提高了马鲛鱼加工副产物的利用率，又为新型铁补充剂的开发提供了技术支持，具有广阔的应用前景。
Description
技术领域
本发明涉及生物技术领域，尤其涉及一种具有强铁螯合能力的马鲛鱼蛋白来源的铁螯合肽及其应用。
背景技术
马鲛鱼为近海中上层鱼类，每年的产量都相当丰富，不仅是浙江更是我国重要的海洋经济鱼类之一。在浙江舟山群岛渔场设有专门马鲛鱼渔场，从东海区主要渔业资源利用状况分析中可以发现马鲛鱼产量开发还有着潜力可以挖掘。据联合国粮农组织统计2015年世界马鲛鱼年捕获量就达149万吨，而且呈上升趋势。作为低值高产鱼的一种，马鲛鱼鱼肉口感微酸，而且肉质相对比较粗糙，但是它的不饱和脂肪酸和蛋白含量非常高，DHA、EPA等的不饱和脂肪酸含量仅次于鲔鱼，而且它还具有提神和防衰老等食疗功能。经常食用对治疗贫血、预防早衰以及神经衰弱等症有一定辅助疗效，因此，马鲛鱼通常用来加工制备鱼油产品，但是，仅仅用来制备鱼油产品会产生大量的加工副产物，约占到总重量的40-60%，这些副产物蛋白质含量高、水分多，不易保存，容易腐烂变质发臭，目前对马鲛鱼加工副产物的利用主要集中在加工成动物饲料和鱼糜，这些加工产物特别是前者附加值相对比较低。这些含有丰富蛋白质资源的加工副产物是生物活性肽的优质原料，但目前未得到有效高值化利用。
铁作为必需微量元素之一在人体中起着非常重要的作用。但是由于许多天然食物中存在植酸及人体肠道呈碱性而使食物中无机铁吸收率较低，近年来的研究表明，一些生物活性小肽与铁的螯合物具有比无机铁更高的吸收效率而且没有副作用，而吸收率相对较高的血红素铁由于呈红色，食品中添加会影响品质，因此，近年来生物活性小肽铁成为研究热点，而如何获得具有铁螯合活性的肽，成为制备新型铁补充剂迫切的研究方向。
发明内容
本发明为了克服马鲛鱼加工副产物利用率低和传统铁补充剂吸收率较低的问题，提供了一种具有强铁螯合能力的马鲛鱼蛋白来源的铁螯合肽。
本发明还提供了一种马鲛鱼蛋白来源的铁螯合肽在铁补充剂中的应用。
为了实现上述目的，本发明采用以下技术方案：
一种马鲛鱼蛋白来源的铁螯合肽，所述马鲛鱼蛋白来源的铁螯合肽的氨基酸序列为Gln-Lys-Gly-Thr-Tyr-Asp-Asp-Tyr-Val-Glu-Gly-Leu，
作为优选，所述马鲛鱼蛋白来源的铁螯合肽的分子量为1387.47 Da。
作为优选，所述马鲛鱼蛋白来源的铁螯合肽的铁螯合能力为824μg/g。
一种马鲛鱼蛋白来源的铁螯合肽的应用，将所述马鲛鱼蛋白来源的铁螯合肽与铁离子溶液螯合，制得铁-肽螯合物，应用于铁补充剂。
本发明马鲛鱼蛋白来源的铁螯合肽具有安全无毒副作用、促进铁吸收特点，其原理为：利用马鲛鱼蛋白来源的铁螯合肽具备与铁离子螯合的作用位点，能够形成稳定多肽-铁螯合物，由于其具有独特的螯合机制和转运机制，易被人体吸收。
作为优选，所述铁补充剂为具有补铁功能的食品、药品和保健品。
因此，本发明具有如下的有益效果：铁螯合能力强，可应用于铁补充剂，安全无毒副作用、促进铁吸收，既提高了马鲛鱼加工副产物的利用率，又为新型铁补充剂的开发提供了技术支持，具有广阔的应用前景。
附图说明
图1是本发明马鲛鱼蛋白来源的铁螯合肽的凝胶过滤层析色谱图。
图2是图1中两个主要组分的铁螯合能力图。
图3是本发明马鲛鱼蛋白来源的铁螯合肽的反相高效液相色谱图。
具体实施方式
下面通过具体实施例并结合附图，对本发明的技术方案作进一步具体的说明。
在本发明中，若非特指，所有设备和原料均可从市场购得或是本行业常用的，下述实施例中的方法，如无特别说明，均为本领域常规方法。
（1）马鲛鱼蛋白质预处理：取马鲛鱼（Scomberomorus niphonius）提取油脂后的加工副产物，按100g物料加入500mL异丙醇，用组织捣碎机打成匀浆，然后于60oC水浴加热，回流，萃取6h，抽滤，得滤渣，于60oC真空干燥，再粉碎机粉碎，过100目筛，得到脱脂后的马鲛鱼加工副产物蛋白，密封保存，备用；
（2）酶促水解：以脱脂后的马鲛鱼加工副产物蛋白为原料，按固液比10g:200mL加入20mM pH8.0 Tris-HCl缓冲液，混匀，于水浴中50oC保温10min。加入占总体积1.0%的碱性蛋白酶（6.0×105U/g），搅拌（150rpm）状态下水解1.5h。将酶促水解液于95oC水浴中加热10min，使酶失活。冷却到温室后，离心（10000rpm，20min，4oC），上清液待等下一步分离；
（3）铁螯合肽的制备：将上清液过0.45μm滤膜，然后过3KDa超滤膜，收集小于3KDa组分，将此组分依次经（a）亲和层析、（b）离子交换层析、（c）凝胶过滤层析和（d）反相高效液相色谱（RP-HPLC）纯化，每一步测定收集峰的铁螯合能力，以铁螯合能力最强的峰进入下一步分离纯化，最后得到铁螯合肽。具体步骤如下：
（a）亲和层析：使用IMAC Sepharose Fast Flow（IMAC-FF）树脂填充的色谱柱，用于分离铁结合肽。在IMAC-FF（5mL）填充柱中加入10mL 100mM FeCl3溶液。用15mL超纯水将未结合的铁洗出柱之后，再用15mL 50mM pH3.6乙酸钠-乙酸缓冲液除去非特异性结合铁。随后，将2mL超滤后的组分装载到柱上。用平衡缓冲液（50mM pH3.6乙酸钠-乙酸）冲洗约15mL，然后用15mL 100mM pH4.5 NaH2PO4溶液洗脱，并收集流出液，冷冻干燥；
（b）离子交换层析：将亲和层析得到的冷冻干燥固体100mg溶于10mL超纯水，并过0.22μm滤膜，上样至DEAE Sepharose Fast Flow阴离子交换柱（1.6×30cm），进行线性梯度洗脱，120min内由A液（pH7.0 20mM磷酸缓冲液）过滤到B液（pH7.0 20mM磷酸缓冲液，含1MNaCl），设定流速1.0mL/min，进样量为1.0mL，洗脱方法为0-120min线性洗脱，由100% A过渡到100% B。紫外280nm监测，得到五个峰，收集这五个吸收峰的流出液，并以菲洛嗪比色法测定铁螯合能力，第2个峰组分（SPB-2）铁螯合能力最强，达到329μg/g；
（c）凝胶过滤层析：将离子交换层析得到的SPB2组分上样（1.0mL）到凝胶过滤层析柱（填料为Superdex G75，柱子为1.0×30cm），以超纯水为洗脱液，设定流速1.0mL/min，紫外214m监测，凝胶过滤层析色谱图如图1所示，得到两个峰SPB-2-1和SPB-2-2。收集这两个吸收峰的流出液，并以菲洛嗪比色法测定铁螯合能力，结果如图2所示，第2个峰组分（SPB-2-2）铁螯合能力最强，达到487μg/g。反复多次，制备约50mL SPB-2-2样品液，并冷冻干燥；
（d）RP-HPLC纯化：将凝胶过滤层析得到的SPB-2-2样品用超纯水配成100ug/mL溶液，利用反相高效液相色谱进行分离，反相色谱柱为C18柱（5μm，4.6 mm×250 mm），进样量20uL，柱温30oC，流速0.8mL/min，流动相A为含0.1%三氟乙酸的5%乙腈，流动相B为含0.1%三氟乙酸的50%乙腈，洗脱梯度为0-5min 100% A流动相，5-60min由100% A线性变化到100%B。检测波长214nm。反相高效液相色谱图如图3所示，按峰收集各组分，共得到a和b两个组分，以菲洛嗪比色法测定铁螯合能力，测得第2个峰组分（b）的铁螯合能力最强，达到824μg/g，组分b即为本发明马鲛鱼蛋白来源的铁螯合肽；
（4）结构鉴定：对本发明马鲛鱼蛋白来源的铁螯合肽以ESI-MS测定，其分子量为1387.47 Da，用蛋白/多肽序列分析仪测定，其氨基酸序列为Gln-Lys-Gly-Thr-Tyr-Asp-Asp-Tyr-Val-Glu-Gly-Leu（QKGTYDDYVEGL）。
一种马鲛鱼蛋白来源的铁螯合肽的应用，将马鲛鱼蛋白来源的铁螯合肽与铁离子溶液螯合，制得铁-肽螯合物，应用于具有补铁功能的食品、药品和保健品中，作为铁补充剂应用。利用马鲛鱼蛋白来源的铁螯合肽具备与铁离子螯合的作用位点，能够形成稳定多肽-铁螯合物，由于其具有独特的螯合机制和转运机制，易被人体吸收，具有安全无毒副作用、促进铁吸收的特点。
以上所述仅为本发明的较佳实施例，并非对本发明作任何形式上的限制，在不超出权利要求所记载的技术方案的前提下还有其它的变体及改型。
序列表
<110> 中国计量大学
<120> 一种马鲛鱼蛋白来源的铁螯合肽及其应用
<140> 2017109805211
<141> 2017-10-19
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 12
<212> PRT
<213> 马鲛鱼(Scomberomorus niphonius)
<400> 1
Gln Lys Gly Thr Tyr Asp Asp Tyr Val Glu Gly Leu
1 5 10
Claims (4)
1.一种马鲛鱼蛋白来源的铁螯合肽，其特征在于，所述马鲛鱼蛋白来源的铁螯合肽的氨基酸序列为Gln-Lys-Gly-Thr-Tyr-Asp-Asp-Tyr-Val-Glu-Gly-Leu。
2.根据权利要求1所述的一种马鲛鱼蛋白来源的铁螯合肽，其特征在于，所述马鲛鱼蛋白来源的铁螯合肽的铁螯合能力为824μg/g。
3.一种如权利要求1所述的马鲛鱼蛋白来源的铁螯合肽的应用，其特征在于，将所述马鲛鱼蛋白来源的铁螯合肽与铁离子溶液螯合，制得铁-肽螯合物，应用于铁补充剂。
4.根据权利要求3所述的一种马鲛鱼蛋白来源的铁螯合肽的应用，其特征在于，所述铁补充剂为具有补铁功能的食品、药品和保健品。
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2231171A2 (en) * | 2007-12-14 | 2010-09-29 | Hofseth Biocare AS | Compositions and methods for increasing iron absorption |
| CN104758925A (zh) * | 2015-04-15 | 2015-07-08 | 浙江海洋学院 | 带鱼鱼骨铁螯合胶原肽的铁螯合用途 |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| EP2231171A2 (en) * | 2007-12-14 | 2010-09-29 | Hofseth Biocare AS | Compositions and methods for increasing iron absorption |
| CN104758925A (zh) * | 2015-04-15 | 2015-07-08 | 浙江海洋学院 | 带鱼鱼骨铁螯合胶原肽的铁螯合用途 |
Non-Patent Citations (2)
| Title |
|---|
| Iron homeostasis in host defence and inflammation;Ganz T等;《Nat Rev Immunol.》;20150831;全文 * |
| 超滤法制备高抗菌抗氧化活性带鱼蛋白亚铁螯合肽(Fe-HPH)的工艺研究;林慧敏等;《中国食品学报》;20120630;全文 * |
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