方晨璐, 黄峻榕, 任瑞珍, 杨麒, 蒲华寅, 刘树兴. 酶解薯类淀粉适用于电镜观察其颗粒表面及内部结构[J]. 农业工程学报, 2018, 34(22): 306-312. DOI: 10.11975/j.issn.1002-6819.2018.22.038
    引用本文: 方晨璐, 黄峻榕, 任瑞珍, 杨麒, 蒲华寅, 刘树兴. 酶解薯类淀粉适用于电镜观察其颗粒表面及内部结构[J]. 农业工程学报, 2018, 34(22): 306-312. DOI: 10.11975/j.issn.1002-6819.2018.22.038
    Fang Chenlu, Huang Junrong, Ren Ruizhen, Yang Qi, Pu Huayin, Liu Shuxing. Amylases enzymolysis of tuber starch granules for surface and internal structure observation under scanning electron microscopy[J]. Transactions of the Chinese Society of Agricultural Engineering (Transactions of the CSAE), 2018, 34(22): 306-312. DOI: 10.11975/j.issn.1002-6819.2018.22.038
    Citation: Fang Chenlu, Huang Junrong, Ren Ruizhen, Yang Qi, Pu Huayin, Liu Shuxing. Amylases enzymolysis of tuber starch granules for surface and internal structure observation under scanning electron microscopy[J]. Transactions of the Chinese Society of Agricultural Engineering (Transactions of the CSAE), 2018, 34(22): 306-312. DOI: 10.11975/j.issn.1002-6819.2018.22.038

    酶解薯类淀粉适用于电镜观察其颗粒表面及内部结构

    Amylases enzymolysis of tuber starch granules for surface and internal structure observation under scanning electron microscopy

    • 摘要: 为了同步研究淀粉颗粒表面小体和壳层结构,用α-淀粉酶、β-淀粉酶和普鲁兰酶,在室温下单一或复合酶解马铃薯、红薯、木薯淀粉颗粒,用扫描电子显微镜观察酶解颗粒并进行性质测试。研究结果表明,单一酶作用时,只有α-淀粉酶可使3种薯类淀粉显露颗粒表面小体(直径29~73 nm)和壳层结构(厚度150~400 nm);马铃薯淀粉的酶解率(1.1%)远低于其他2种淀粉的(14.1%、16.3%)。马铃薯淀粉表面小体的排列较紧密、壳层结构较致密,决定了其具有较强的抗酶解性和较大的峰值黏度(即膨胀能力)。复合酶作用时,α-淀粉酶复合与其单一作用的效果类似。因此单一α-淀粉酶有限酶解法可以作为淀粉颗粒表面小体和壳层结构的研究方法,酶解条件为:酶浓度80 U/mL,室温下酶解12 h。该研究结果为淀粉类产品在实际加工过程中的品质控制提供了理论基础。

       

      Abstract: Research on starch structure can provide a theoretical basis for the modification reaction and application of starch. The surface blocklets and shell structure of starch granules determine their enzymolysis and swelling characteristics. At room temperature, three kinds of amylase (α-amylase, β-amylase and pullulanase) were used alone or in compound for three tuber starch (potato, sweet potato and cassava starch) hydrolysis respectively. The enzymolysis rate of starch granules was calculated, gelatinization enthalpy and relative crystallinity of three tuber starch granules before and after limited enzymolysis were calculated by differential scanning calorimetry and X-rays diffraction, respectively. The surface blocklets and shell structure of the enzymolyzed starch granules was observed by scanning electron microscopy. The results showed that the enzymolysis rate of α-amylase was higher than β-amylase and pullulanase. The non-reducing end and α-1,6 glucoside bond located mainly inside, while α-1,4 glucosidic bond located mainly in surface of starch granules. The gelatinization enthalpy and relative crystallinity of the enzymolyzed starch granules increased slightly, these physicochemical properties were close to those of the native starches. The results indicated that the structure of starch granules after enzymolysis was similar to that of native starch granules. Potato starch showed much lower enzymolysis rate (1.1%) than the other two starches (14.1%, 16.3%), and it had the strongest resistance to enzymolysis.In the reaction of single amylase, α-amylase could make the surface of three tuber starch granules appear scratches and cracks, and expose surface blocklets (diameter 29-73 nm) and inner shell structure (thickness 150-400 nm). The surface blocklets of potato starch granules were tightly packed; while those of sweet potato starch granules were loosely arranged with holes, and those of cassava starch granules showed collapses and cracks. The enzymolysis rate of potato starch was much lower than that of the other two starches. Potato starch had strong resistance to enzymes. The results indicated that structure of surface blocklets determined the enzymolysis characteristics of starch granules. Compared with sweet potato and cassava starches, the shell structure of potato starch was dense and orderly. This was consistent with the results of peak viscosity measurement. The pasting properties of three tuber starches (5%, w/w) were measured by using rapid viscosity analyzer (RVA), and the peak viscosity of potato starch (2216 mPa·s) was significantly higher than those of sweet potato and cassava starches (593 and 740 mPa·s). Potato starch granules swelled to a larger degree, indicating that the shell structure determines the expansion characteristics of starch.After β-amylase hydrolysis, only surface bolcklets of granules was observed. After pullulanase treatment, there was no obvious change for granules and no surface blocklets or shell were observed. When hydrolyzed with mixed amylase, the composite reaction of α-amylase was similar to that of single reaction. For example, after hydrolysis with combination of α-amylase and β-amylase, surface blocklets with sizes of 36-65, 32-61 and 32-52 nm appeared on the surface of potato, sweet potato and cassava starches, respectively. Potato and sweet potato starch showed shell structure (thickness 300-400 nm and 100-150 nm, respectively). Sweet potato and cassava starch appeared hollow shell structure (thickness 100-150 nm and 100-200 nm, respectively). Treatment with β-amylase or pullulanase, alone or mixed, could not show shell or outer shell structure for three tuber starch granules. Therefore, the single amylase hydrolysis of α-amylase (80 U/mL, room temperature, 12 h) could be used as a method to study surface blocklets and shell structure of starch granules simultaneously.

       

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