蜡质玉米、稻米抗性淀粉的酶法制备及性质研究
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摘要
通过普鲁兰酶对蜡质玉米和蜡质稻米淀粉脱支后凝沉得到系列抗性淀粉样品。采用淀粉-碘复合物吸光度值法、扫描电镜(SEM)、X-射线衍射、差示量热扫描仪(DSC)、凝胶渗透色谱(GPC)和In-Vitro体外消化模型对抗性淀粉的直链淀粉含量、颗粒形貌、结晶性质、热性质、分子量分布和体外消化性能等进行了深入研究。
(1)通过对蜡质玉米和蜡质稻米淀粉糊化,在普鲁兰酶作用下对淀粉糊进行脱支,然后在低温环境下凝沉,来提高淀粉中的抗性淀粉含量。对蜡质玉米淀粉,在普鲁兰酶浓度20ASPU/g(淀粉干基重),脱支24h,25℃条件下放置24h后,RS含量达到最大值27.69%。对蜡质稻米淀粉,在普鲁兰酶浓度55ASPU/g(淀粉干基重),脱支12h,25℃条件下放置24h后,RS含量达到最大值28.61%。与原淀粉(蜡质玉米淀粉和蜡质稻米淀粉的RS含量分别为1.30%和0.33%)相比,淀粉通过糊化、酶脱支、凝沉处理后,RS含量增加。
(2)研究了蜡质玉米和蜡质稻米抗性淀粉的各种性质。采用淀粉-碘复合物吸光度值法测定直链淀粉含量,发现经过普鲁兰酶处理,样品的直链淀粉含量都高于原淀粉的直链淀粉含量。一般来说,随着直链淀粉含量的增大,相应的抗性淀粉含量也大。扫描电镜照片显示当淀粉糊经过脱支、凝沉、干燥后,形成了大小不等,颗粒不规则的碎片。X-射线衍射表明所有的样品显示相似的强衍射峰,在经过脱支、凝沉处理之后显示B和V型混合结晶结构,抗性淀粉样品的结晶度有了较大的提高。一般结晶度高的样品,其抗性淀粉含量也高。与原淀粉相比,淀粉经过处理后内部的结构变得紧密坚固,而且还形成了一定数量的新晶体。抗性淀粉的热力学性质参数To、Tp、Tc、Tc- To以及焓值△H都有不同程度的提高。从分子量分布图谱可以看出,抗性淀粉的分子量大大降低。
(3)采用In-Vitro消化模型,分别在不同消化时间内对经过普鲁兰酶脱支的蜡质玉米和蜡质稻米抗性淀粉样品的消化产物和平均消化速度进行测定,研究抗性淀粉样品在经酶解脱支、凝沉处理之后消化性能的变化规律。由于猪胰α-淀粉酶的作用方式、A型和B型结晶结构和双螺旋结构的紧密程度等因素的不同导致了原淀粉和抗性淀粉在消化性能方面的不同。在前3个小时中,因为在反应前期α-淀粉酶的浓度大,效率高,所有样品的消化产物的量升高,消化速率都急速下降。在反应3小时后,淀粉分子被水解成过短的分子链,淀粉分子和的结合位点减少,消化产物的量增加不明显,消化速率缓慢降低。
Waxy corn starch and waxy rice starch were used as material to prepare a series of resistant starch samples by pullulanase in this dissertation. The amylose content, particle morphology, crystalline properties, thermal properties, molecular weight distribution and in vitro digestibility of resistant starch samples were investigated penetratingly by the method of starch - iodine complex absorbance value, scanning electron microscopy, X-ray diffraction, DSC, GPC, and in-vitro digestion model.
Firstly, the waxy corn and waxy rice starch pastes were treated by pullulanase, and then retrograded at low temperature to improve the resistant starch formation. When concentration of pullulanase was 20ASPU/g(dry starch weight), debranched time was 24h and stored 24h at 25℃, the content of waxy maize resistant starch was 27.69%. When concentration of pullulanase was 55ASPU/g(dry starch weight), debranched time was 12h and stored 24h at 25℃, the content of waxy rice resistant starch was 28.61%. Compared with waxy maize and waxy rice native starches (RS content was 1.30% and 0.33%, respectively), RS content of samples increased observably.
Secondly, various properties of waxy corn and waxy rice resistant starches were determined. The amylose content of samples was studied by the method of starch - iodine complex absorbance value. After treatment with pullulanase, the amylose content of samples was higher than that of native starches. In general, with increasing amylose content, resistant starch content increased correspondly. SEM photos showed that starch granule appearance changed to different size particles of irregular fragments and structure disappeared. Compared with native starches, crystal structure of samples changed from A type to a mixture of B and V–type, besides, the crystallinity of resistant starch also improved. The higher crystallinity of starch, the higher RS content. DSC determination showed that To、Tp、Tc and△H all increased, and these change had good relation with RS content. The molecular weight distribution of resistant starches significantly reduced.
Thirdly, the total carbohydrates digestion products and the average rate of digestion of resistant starches were discussed. With increasing incubation time, the total carbohydrates digestion products were increased and the average rates of digestion were decreased. In the first 3 hours, the digestion products of all samples and the average rate of digestion were sharply increasing. This was mainly attributed to the high concentration and efficiency of pancreaticα-amylase solution in the former reaction. After 3h, starch molecules were hydrolyzed into small molecular chain and the binding sites between starch molecules andα-amylase were decreased, so the digestion of resistant starch slowed down. The longer the substrate–enzyme contact time, the more starch was digested within the duration of the experiment.
(1)通过对蜡质玉米和蜡质稻米淀粉糊化,在普鲁兰酶作用下对淀粉糊进行脱支,然后在低温环境下凝沉,来提高淀粉中的抗性淀粉含量。对蜡质玉米淀粉,在普鲁兰酶浓度20ASPU/g(淀粉干基重),脱支24h,25℃条件下放置24h后,RS含量达到最大值27.69%。对蜡质稻米淀粉,在普鲁兰酶浓度55ASPU/g(淀粉干基重),脱支12h,25℃条件下放置24h后,RS含量达到最大值28.61%。与原淀粉(蜡质玉米淀粉和蜡质稻米淀粉的RS含量分别为1.30%和0.33%)相比,淀粉通过糊化、酶脱支、凝沉处理后,RS含量增加。
(2)研究了蜡质玉米和蜡质稻米抗性淀粉的各种性质。采用淀粉-碘复合物吸光度值法测定直链淀粉含量,发现经过普鲁兰酶处理,样品的直链淀粉含量都高于原淀粉的直链淀粉含量。一般来说,随着直链淀粉含量的增大,相应的抗性淀粉含量也大。扫描电镜照片显示当淀粉糊经过脱支、凝沉、干燥后,形成了大小不等,颗粒不规则的碎片。X-射线衍射表明所有的样品显示相似的强衍射峰,在经过脱支、凝沉处理之后显示B和V型混合结晶结构,抗性淀粉样品的结晶度有了较大的提高。一般结晶度高的样品,其抗性淀粉含量也高。与原淀粉相比,淀粉经过处理后内部的结构变得紧密坚固,而且还形成了一定数量的新晶体。抗性淀粉的热力学性质参数To、Tp、Tc、Tc- To以及焓值△H都有不同程度的提高。从分子量分布图谱可以看出,抗性淀粉的分子量大大降低。
(3)采用In-Vitro消化模型,分别在不同消化时间内对经过普鲁兰酶脱支的蜡质玉米和蜡质稻米抗性淀粉样品的消化产物和平均消化速度进行测定,研究抗性淀粉样品在经酶解脱支、凝沉处理之后消化性能的变化规律。由于猪胰α-淀粉酶的作用方式、A型和B型结晶结构和双螺旋结构的紧密程度等因素的不同导致了原淀粉和抗性淀粉在消化性能方面的不同。在前3个小时中,因为在反应前期α-淀粉酶的浓度大,效率高,所有样品的消化产物的量升高,消化速率都急速下降。在反应3小时后,淀粉分子被水解成过短的分子链,淀粉分子和的结合位点减少,消化产物的量增加不明显,消化速率缓慢降低。
Waxy corn starch and waxy rice starch were used as material to prepare a series of resistant starch samples by pullulanase in this dissertation. The amylose content, particle morphology, crystalline properties, thermal properties, molecular weight distribution and in vitro digestibility of resistant starch samples were investigated penetratingly by the method of starch - iodine complex absorbance value, scanning electron microscopy, X-ray diffraction, DSC, GPC, and in-vitro digestion model.
Firstly, the waxy corn and waxy rice starch pastes were treated by pullulanase, and then retrograded at low temperature to improve the resistant starch formation. When concentration of pullulanase was 20ASPU/g(dry starch weight), debranched time was 24h and stored 24h at 25℃, the content of waxy maize resistant starch was 27.69%. When concentration of pullulanase was 55ASPU/g(dry starch weight), debranched time was 12h and stored 24h at 25℃, the content of waxy rice resistant starch was 28.61%. Compared with waxy maize and waxy rice native starches (RS content was 1.30% and 0.33%, respectively), RS content of samples increased observably.
Secondly, various properties of waxy corn and waxy rice resistant starches were determined. The amylose content of samples was studied by the method of starch - iodine complex absorbance value. After treatment with pullulanase, the amylose content of samples was higher than that of native starches. In general, with increasing amylose content, resistant starch content increased correspondly. SEM photos showed that starch granule appearance changed to different size particles of irregular fragments and structure disappeared. Compared with native starches, crystal structure of samples changed from A type to a mixture of B and V–type, besides, the crystallinity of resistant starch also improved. The higher crystallinity of starch, the higher RS content. DSC determination showed that To、Tp、Tc and△H all increased, and these change had good relation with RS content. The molecular weight distribution of resistant starches significantly reduced.
Thirdly, the total carbohydrates digestion products and the average rate of digestion of resistant starches were discussed. With increasing incubation time, the total carbohydrates digestion products were increased and the average rates of digestion were decreased. In the first 3 hours, the digestion products of all samples and the average rate of digestion were sharply increasing. This was mainly attributed to the high concentration and efficiency of pancreaticα-amylase solution in the former reaction. After 3h, starch molecules were hydrolyzed into small molecular chain and the binding sites between starch molecules andα-amylase were decreased, so the digestion of resistant starch slowed down. The longer the substrate–enzyme contact time, the more starch was digested within the duration of the experiment.
引文
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