CN108424942B - 一种葡糖基壳核结构的载体材料及其制备与应用 - Google Patents
一种葡糖基壳核结构的载体材料及其制备与应用 Download PDFInfo
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Abstract
本发明公开了一种葡糖基壳核结构的载体材料及其制备与应用,属于现代食品加工技术领域。本发明以球状超支化的水溶性淀粉粒子为原料,采用酶法接枝扩链的工艺处理,将水溶性葡聚糖分子表面改性成具有结晶结构密集堆积的坚固壳结构,使之形成具有“内核腔无定形态、外壳层结晶态”葡糖基壳核结构载物材料。本发明所采用的球状超支化的水溶性淀粉粒子,原料来源广、不受产地和季节的限制;步骤简便,易于操作,反应条件可控,成本相对较低,对环境基本无污染;本发明制备的产品有效对功能性营养成分的保护、传输及释放,可应用于食品、医药、日用化学品等多个领域,市场前景十分看好,经济效益广阔。
Description
技术领域
本发明涉及一种葡糖基壳核结构的载体材料及其制备与应用,属于现代食品加工技术领域。
背景技术
随着社会科技和经济的发展、生活方式的转化及环境的恶化,我国疾病谱发生变化,糖尿病、高血压、肥胖症等慢性病数量剧增,亚健康状态的人群也越来越庞大。与此同时,人们的健康保健意识也越来越强烈,医疗观念已由病后治疗型向预防保健型转变。通过健康的生活方式、借助膳食营养等手段预防慢性疾病,正越来越被人们重视。
功能性食品由于其显著的生理功能已成为消费者食谱中预防或减少慢性疾病发生的重要途径。然而,许多天然活性组分存在熔点高、水溶性差、易光解、易氧化分解、不易被人体消化吸收等特性,因此设计有效的活性组分传递系统来提高生物活性物质的利用率正成为该领域的研究重点。目前,罗氏制药、帝斯曼、巴斯夫等国外主要天然营养素生产供应商都开发了天然营养素的微胶囊及乳液保护技术并形成了规模化的生产销售。基于此,本发明对一种提高脂溶性组分溶解度与生物利用度的加工方法进行了详细研究。
当今,环境保护,资源节约已经成为世界各国实现可持续发展的基本战略。在全球资源供给紧张、环境问题日益突出,低碳经济发展需求日益强烈,可再生资源为原料的生物载物材料必将迅猛发展与广泛应用。以高分子物质为原料转化制造的生物载物材料作为国际战略性新兴产业。目前,蛋白质生物大分子在药学领域的应用发展很快,比如YeonheeYund等人研究发现了口服的靶向蛋白纳米粒药物载体。但这类药物载体的在胃肠稳定性相对较差;易变性,不易吸收,影响了它们的生物利用度。然而,以淀粉基为原料的载物材料中,例如:张根义、杨英等人在CN101293998中公开了一种水溶性纳米功能性脂肪酸复合物载体的制备方法。但得到的载物复合物产率较低,原料损耗较多,且制作工艺较为复杂。
发明内容
为了解决上述问题,本发明提供了一种可包埋功能性营养成分的葡聚糖基壳核结构载体材料及其加工方法。本发明采用可溶性的淀粉粒子,通过生物技术改性使之形成具有特殊的“内核腔无定形态、外壳层结晶态”壳核结构,在此过程中,通过“藤缠绕”的方法载运营养因子。本发明的葡聚糖基壳核结构载体材料,能够提高功能活性成分的生物稳定性、生物利用有效性及缓释功效。本发明的加工方法具有生产工艺简单、产率高、技术先进、安全性高、可保护和调节功能性营养成分的输送与释放等特点。
本发明的第一个目的是提供一种可包埋功能性营养成分的葡聚糖基壳核结构载体材料,所述葡聚糖基壳核结构载体材料,包括利用糖基转移酶将葡萄糖基以α-1,4糖苷键,在球状超支化的水溶性淀粉粒子的外表面进行接枝扩链得到的。
在一种实施方式中,所述水溶性淀粉粒子分子量107-108g/mol,α-1,6糖苷键比例为7%-10%,平均粒径30-100nm。
在一种实施方式中,所述球状超支化的水溶性淀粉粒子可以是来源于天然植物球状超支化淀粉颗粒、动物中(牡蛎糖原)、生物技术合成高分子球状多糖等等。
在一种实施方式中,所述球状超支化的水溶性淀粉粒子为来源于甜质型可溶性玉米葡聚糖。
在一种实施方式中,所述糖基转移酶可以采用糖原磷酸化酶、α-葡萄糖磷酸化酶等。
在一种实施方式中,所述包埋功能性营养成分包括亚油酸、亚麻酸、Q10等等。
在一种实施方式中,所述接枝扩链的反应体系中还含有功能性成分。
在一种实施方式中,所述葡聚糖基壳核结构载体材料的制备,是先将水溶性淀粉粒子配置成溶液,然后在含有提供葡萄糖分子的供体分子和糖基转移酶的体系中进行反应,反应后经灭酶、离心、干燥沉淀,得到葡聚糖基壳核结构载体材料。
在一种实施方式中,所述提供葡萄糖分子的供体分子与水溶性淀粉粒子的质量比为1.5:1~5:1。
在一种实施方式中,所述提供葡萄糖分子的供体分子为葡萄糖-1-磷酸盐。
在一种实施方式中,所述葡萄糖-1-磷酸盐可以选用钠盐或钾盐。
在一种实施方式中,所述制备,是以来源于甜质型可溶性玉米葡聚糖为主要原料,利用糖基转移酶将球状超支化的玉米葡聚糖接枝扩链,进而形成“内核腔无定形态、外壳层结晶态”的葡糖基壳核结构的载物材料。
在一种实施方式中,所述制备,具体加工步骤:
(1)将水溶性淀粉粒子溶于缓冲液配制成质量浓度为0.5~3.0%的均一溶液;
(2)按照每1g水溶性淀粉粒子添加1.5~5g葡萄糖-1-磷酸盐和10~180U糖基转移酶的比例,添加葡萄糖-1-磷酸盐和糖基转移酶,搅拌均匀,在温度35~40℃,pH 6.5~7.5条件下恒温反应3~24h;
(3)加热灭酶和离心处理,将所获沉淀物真空干燥,即得到葡糖基壳核结构载体材料。
在一种实施方式中,所述缓冲液为Tris-HCl缓冲液,可选地,缓冲液50mmol/L、pH7.0。
本发明的第二个目的是提供一种包埋功能性成分的复合物,所述复合物是在本发明葡聚糖基壳核结构载体材料形成过程中,往反应体系中加入功能性成分制备得到的。
在一种实施方式中,所述功能性成分可以营养成分,比如亚油酸、亚麻酸、Q10等。
本发明的第三个目的是提供一种生物载物材料,含有本发明的葡聚糖基壳核结构载体材料。
本发明的第四个目的是提供所述葡聚糖基壳核结构载体材料在载物方面的应用。
在一种实施方式中,所述载物运载的是药物或者功能性营养成分。
本发明的第五个目的是提供所述葡聚糖基壳核结构载体材料在食品、医药、日用化学品等领域的应用,包括但不限于功能因子靶向控释、纳米粒包埋等等。
本发明的优点和效果:
本发明以球状超支化的水溶性淀粉粒子为原料,采用酶法接枝扩链的工艺处理,将水溶性葡聚糖分子表面改性成具有结晶结构密集堆积的坚固壳结构,使之形成具有“内核腔无定形态、外壳层结晶态”葡糖基壳核结构载物材料,具有如下优点:
(1)本发明所采用的球状超支化的水溶性淀粉粒子,可以充分利用我国资源丰富的谷物原料,原料来源广、不受产地和季节的限制,而且原料具有生物可降解性,环保;尤其是天然玉米多糖,其具备优良生物相容性,更适用于医药领域。
(2)本发明步骤简便,易于操作,反应条件可控,成本相对较低,而且采用清洁绿色生产工艺,对环境基本无污染。
(3)本发明制备的产品有效对功能性营养成分的保护、传输及释放。可应用于食品、医药、日用化学品等多个领域,如功能因子靶向控释、纳米粒包埋等等,市场前景十分看好,经济效益广阔。
附图说明
图1为具“壳核结构”载物材料及其与营养因子复合物的示意图;
图2为原淀粉粒子与具“壳核结构”载物材料的X射线衍射图结果。
具体实施方案
为了更好地实现本发明专利,生物稳定性通过测定其氧化率POV(过氧化值)表征,应用硫氰酸钾POV的测定方法。其公式为:其中,c与c0为测试样品和样品空白中铁的质量;m为CLA的质量;2是转换因子;55.84是铁的相对原子质量。其中以纯营养因子的氧化程度作为对照,计算并比较对照与材料中的过氧化物量(POV)的最大值。其中,POV为脂肪氧化后得到第一阶段产物过氧化物量。由于不能排除少量过氧化物在氧化环境下继续分解成小分子物质,因此稳定性表征为:
稳定性≤(100-100gCLA氧化产生的过氧化物量最大值)/100×100%
细胞实验:对载物复合物做肠道细胞实验。将细胞培养液中添加100μL载物材料-营养因子复合物溶解液,加入2mM双氧水2h刺激细胞,细胞在继续培养4h。采用MTT法检测细胞活力。
下面结合实例进一步阐明本发明的内容,但本发明所保护的内容不仅仅局限于下面的实例。
实施例1
称取1g水溶性淀粉粒子溶于Tris-HCl缓冲液(50mmol/L,pH7.0)配制成质量浓度为0.5%的均一溶液。继续添加1.5g葡萄糖-1-磷酸盐和40U糖基转移酶,搅拌均匀,在温度40℃,pH7.0条件下恒温反应12h。加热灭酶和离心处理,将所获沉淀物真空干燥,即得到葡糖基壳核结构载体材料。
如图1所示,a表示球状超支化水溶性淀粉粒子;b表示淀粉粒子经过生物技术改性,接枝扩链的初期阶段,即糖基转移酶将葡萄糖基以α-1,4糖苷键依次连接到球状淀粉颗粒的非还原性末端。c表示淀粉粒子经过生物技术改性,接枝扩链的后期阶段,即接枝所形成的直链结构在球状淀粉粒子外表面缠绕、交联,部分位置形成双螺旋结构,经堆积聚集,进而形成“内核腔无定形态、外壳层结晶态”壳核结构。d代表着经“藤缠绕”方法得到的载物材料与营养因子复合物,接枝扩链产生的直链单螺旋,因内部疏水、外部亲水,能与营养因子以疏水相互作用包合形成载物材料与营养因子复合物。
图2为原淀粉粒子与具“壳核结构”载物材料的X射线衍射图结果。结果表示从无定型转变为具一定晶体结构。
表1 具壳核结构载物材料性质
实施例2
称取1g水溶性淀粉粒子溶于Tris-HCl缓冲液(50mmol/L,pH7.0)配制成质量浓度为1.0%的均一溶液。继续添加2.5g葡萄糖-1-磷酸盐和60U糖基转移酶,搅拌均匀,在温度40℃,pH7.0条件下恒温反应18h。加热灭酶和离心处理,将所获沉淀物真空干燥,即得到葡糖基壳核结构载体材料。
实施例3
称取1g水溶性淀粉粒子溶于Tris-HCl缓冲液(50mmol/L,pH7.0)配制成质量浓度为1.5%的均一溶液。继续添加5.0g葡萄糖-1-磷酸盐和100U糖基转移酶,搅拌均匀,在温度40℃,pH7.0条件下恒温反应24h。加热灭酶和离心处理,将所获沉淀物真空干燥,即得到葡糖基壳核结构载体材料。
实施例4:葡糖基壳核结构载体材料的应用
将实施例1-3制备得到的葡糖基壳核结构载体材料,用于载运功能性活性物质共轭亚油酸。具体试验方法:
在淀粉粒子接枝扩链的反应进行中加入营养因子,即在将1g淀粉粒子溶于缓冲溶液,相继加入5.0g葡萄糖-1-磷酸盐、100U酶、溶于少量乙醇的10mg营养因子共轭亚油酸使之互混,搅拌均匀,在温度40℃,pH7.0条件下恒温反应24h。完成接枝扩链构成壳核结构,结构因含具疏水作用单螺旋空腔,能包合营养素,进而形成复合物。可以加15%氯化钠溶液加速产物生成,经离心、50%醇洗、干燥处理得到载物复合物。
表2 葡糖基壳核结构载体材料载运共轭亚油酸
生物稳定性 | 生物稳定性提高率 | |
实施例1 | 91.4% | 33.2% |
实施例2 | 94.6% | 36.7% |
实施例3 | 97.3% | 39.1% |
对比例1 | 58.2% | / |
对比例2 | 88.7% | 30.5% |
其中,对比例1是对照组即营养因子——共轭亚油酸。对比例2的实施方法为将直链淀粉在二甲亚砜溶液90℃做溶解处理,冷却至30℃。将其与相同温度含共轭亚油酸的二甲亚砜互混,完成单螺旋-营养因子包合。加20倍体积相同温度的去离子水及15%氯化钠溶液加速产物生成,经离心、50%醇洗、干燥处理得到载物材料-营养因子复合物。
实施例5:葡糖基壳核结构载体材料的应用
将实施例3制备得到的葡糖基壳核结构载体材料,用于载运功能性活性物质辅酶Q10。
具体试验方法:
在淀粉粒子接枝扩链的反应进行中加入营养因子,即在将1g淀粉粒子溶于缓冲溶液,相继加入10.0g葡萄糖-1-磷酸盐、100U酶、溶于少量乙醇的10mg营养因子辅酶Q10使之互混,搅拌均匀,在温度40℃,pH7.0条件下恒温反应24h。完成接枝扩链构成壳核结构,结构因含具疏水作用单螺旋空腔,能包合营养素,进而形成复合物。加15%氯化钠溶液加速产物生成,经离心、50%醇洗、干燥处理得到载物复合物。
表3 肠道细胞实验
本文所描述的具体实施案例仅作为对本发明精神和部分实验做举例说明。本发明所述领域的技术人员可以对所描述的具体实施案例做出各种各样的修改或补充或采用类似的方式替代,但并不会偏离本发明的精神或者超越所附权利要求书所定义的范围。
Claims (6)
1.一种葡聚糖基壳核结构载体材料,其特征在于,所述葡聚糖基壳核结构载体材料是利用糖基转移酶将葡萄糖基以α-1,4糖苷键,在球状超支化的水溶性淀粉粒子的外表面进行接枝扩链得到的;所述葡聚糖基壳核结构为内核腔无定形态、外壳层结晶态的壳核结构;
所述水溶性淀粉粒子分子量107 ~ 108g/mol,α-1,6糖苷键比例为7% ~ 10%,平均粒径30 ~ 100nm;
所述葡聚糖基壳核结构载体材料的制备,具体加工步骤如下:
(1)称取1g水溶性淀粉粒子溶于缓冲液配制成质量浓度为0.5 ~ 3.0%的均一溶液;
(2)按照每1g水溶性淀粉粒子添加1.5 ~ 5g葡萄糖-1-磷酸盐和10 ~ 180U糖基转移酶的比例,添加葡萄糖-1-磷酸盐和糖基转移酶,搅拌均匀,在温度35 ~ 40℃,pH 6.5 ~ 7.5条件下恒温反应3-24 h;
(3)加热灭酶和离心处理,将所获沉淀物真空干燥,即得到葡糖基壳核结构载体材料。
2.根据权利要求1所述的葡聚糖基壳核结构载体材料,其特征在于,所述球状超支化的水溶性淀粉粒子来源于天然植物球状超支化淀粉颗粒、生物技术合成高分子球状多糖。
3.根据权利要求1所述的葡聚糖基壳核结构载体材料,其特征在于,所述球状超支化的水溶性淀粉粒子为来源于甜质型可溶性玉米葡聚糖。
4.一种包埋功能性成分的复合物,其特征在于,所述复合物是在权利要求1~3任一所述的葡聚糖基壳核结构载体材料形成过程中,往反应体系中加入功能性成分制备得到的。
5.一种生物载物材料,含有权利要求1~3任一所述的葡聚糖基壳核结构载体材料。
6.权利要求1~3任一所述的葡聚糖基壳核结构载体材料在载物方面的应用。
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