CN102860538A - Alternate double-frequency countercurrent ultrasonic-assisted simultaneous extraction method for seaweed protein and polysaccharide mixed product - Google Patents
Alternate double-frequency countercurrent ultrasonic-assisted simultaneous extraction method for seaweed protein and polysaccharide mixed product Download PDFInfo
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Abstract
本发明提供紫菜蛋白与多糖混合产品的交替双频逆流超声辅助同时提取方法，涉及海洋产品的开发利用领域。将干紫菜粉碎至40目，向紫菜粉中加10～20倍紫菜粉重量的自来水溶解；调温度为60~100℃，调pH为8.0~10.0；在每升体积超声功率200W的基准条件下，采用交替双频逆流超声辅助水提法进行提取，具体参数为15kHz和20kHz双频交替工作、超声交替工作时间1~5s、提取时间30~80min，料液以逆流循环的方式通过超声探头；提取结束后，离心取上清液，将上清液浓缩、喷雾干燥，得到产品。本方法较常规提取方法提取效率更高；较单频超声辅助提取方法破壁和传质效果更强，能量利用率更高。
The invention provides an alternate dual-frequency countercurrent ultrasonic-assisted simultaneous extraction method for a mixed product of laver protein and polysaccharide, and relates to the field of development and utilization of marine products. Crush dried seaweed to 40 mesh, add 10-20 times the weight of seaweed powder to the seaweed powder to dissolve in tap water; adjust the temperature to 60-100°C, adjust the pH to 8.0-10.0; under the standard condition of ultrasonic power per liter of volume 200W , using alternate dual-frequency countercurrent ultrasonic-assisted water extraction method for extraction, the specific parameters are 15kHz and 20kHz dual-frequency alternate operation, ultrasonic alternate working time 1~5s, extraction time 30~80min, and the feed liquid passes through the ultrasonic probe in the way of countercurrent circulation; After the extraction is completed, the supernatant is obtained by centrifugation, concentrated and spray-dried to obtain the product. Compared with the conventional extraction method, the extraction efficiency of the method is higher; compared with the single-frequency ultrasonic-assisted extraction method, the effect of wall breaking and mass transfer is stronger, and the energy utilization rate is higher.
Description
the
技术领域 technical field
紫菜蛋白与多糖混合产品的交替双频逆流超声辅助同时提取方法，涉及海洋产品的开发利用，属于紫菜精深加工技术领域。 The invention discloses an alternate dual-frequency countercurrent ultrasonic-assisted simultaneous extraction method for a mixed product of laver protein and polysaccharide, relates to the development and utilization of marine products, and belongs to the technical field of deep processing of laver.
技术背景 technical background
条斑紫菜（Porphyra yezoensis）是一种营养价值很高的食用海藻类产品。研究发现，条斑紫菜中蛋白质和总糖含量很高（40.22%和34.07%），是一种优质蛋白质和多糖的生产原料。紫菜蛋白和多糖具有良好的医疗保健功效，如紫菜多糖在降血脂、抗凝血、抗血栓形成、降血糖等方面具有显著的功效；紫菜藻红蛋白是一种重要的生理活性物质。因此，目前人们对紫菜蛋白和多糖作为一种保健食用蛋白和多糖的兴趣日益增加。 Porphyra yezoensis is an edible seaweed product with high nutritional value. The research found that Porphyra zebra has high protein and total sugar content (40.22% and 34.07%), and it is a raw material for the production of high-quality protein and polysaccharides. Laver protein and polysaccharides have good health care effects, such as laver polysaccharides have significant effects in reducing blood fat, anticoagulation, antithrombosis, and hypoglycemia; laver phycoerythrin is an important physiologically active substance. Therefore, there is currently an increasing interest in laver protein and polysaccharides as a health food protein and polysaccharide.
目前大多数紫菜只是简单干燥成型后就直接食用，产品品种单一，而且紫菜产品的价值也没有得到充分地利用，造成紫菜优质蛋白、多糖资源的严重浪费。因此，需要寻找一种紫菜蛋白和多糖的高效提取方法。目前紫菜蛋白和多糖的常规提取方法是碱溶酸沉提取法和水提取法。紫菜蛋白碱溶酸沉提取法会使产品含有大量的无机盐，导致后期脱盐处理成本增加，且提取效率较低。紫菜多糖水提取法的破壁和传质效果较差，能量利用率不高，很难获得较高的提取效率。目前超声辅助提取技术作为替代传统提取技术的应用水平在不断地提升，在紫菜蛋白和多糖提取方面已有应用。超声波可以利用其空化、热和机械效应提高产品的提取效率，但是目前超声辅助提取技术都是采用单一频率的超声模式。对于复杂的紫菜蛋白和多糖结构及存在形式，单一频率的超声很难取得理想的溶解促进效果和传质强化效果，不能很好地达到缩短提取时间和提高提取得率的目的，针对该技术的发展瓶颈，急需寻求一个解决的方法。 At present, most seaweeds are simply dried and shaped to be eaten directly. The product variety is single, and the value of seaweed products has not been fully utilized, resulting in a serious waste of seaweed high-quality protein and polysaccharide resources. Therefore, it is necessary to find a high-efficiency extraction method of laver protein and polysaccharide. At present, the conventional extraction methods of laver protein and polysaccharides are alkali-soluble acid precipitation extraction and water extraction. Alkali-soluble and acid-precipitation extraction of laver protein will cause the product to contain a large amount of inorganic salts, resulting in increased cost of desalination in the later stage and low extraction efficiency. The water extraction method of Porphyra polysaccharides has poor wall breaking and mass transfer effects, and the energy utilization rate is not high, so it is difficult to obtain a high extraction efficiency. At present, the application level of ultrasonic-assisted extraction technology as an alternative to traditional extraction technology is constantly improving, and it has been applied in the extraction of laver protein and polysaccharides. Ultrasound can improve the extraction efficiency of products by using its cavitation, thermal and mechanical effects, but the current ultrasonic-assisted extraction technology uses a single-frequency ultrasonic mode. For the complex structures and forms of laver protein and polysaccharides, it is difficult to achieve the ideal effect of dissolution promotion and mass transfer enhancement by a single frequency of ultrasound, and it cannot well achieve the purpose of shortening the extraction time and improving the extraction yield. The development bottleneck urgently needs to find a solution. the
发明内容 Contents of the invention
本发明为了克服上述现有技术的不足，提供一种紫菜蛋白与多糖混合产品的交替双频逆流超声辅助同时提取方法。 In order to overcome the deficiencies of the above-mentioned prior art, the present invention provides an alternate dual-frequency countercurrent ultrasonic-assisted simultaneous extraction method of a laver protein and polysaccharide mixed product.
本发明所述的紫菜蛋白与多糖混合产品的交替双频逆流超声辅助同时提取方法，按照下述步骤进行： The alternate double-frequency countercurrent ultrasonic-assisted simultaneous extraction method of the laver protein and polysaccharide mixed product according to the present invention is carried out according to the following steps:
（1）将干紫菜片粉碎至40目，向紫菜粉中加10~20倍紫菜粉重量的自来水溶解；调温度为60~100℃，调pH为8.0~10.0； (1) Crush the dried laver flakes to 40 mesh, add 10-20 times the weight of laver powder to the laver powder to dissolve in tap water; adjust the temperature to 60-100°C, and adjust the pH to 8.0-10.0;
（2）在每升体积超声功率200W的基准条件下，采用交替双频逆流超声辅助提取，频率为15 kHz和20 kHz的聚能式超声波探头相互呈一定角度安装，料液自下而上以逆流循环的方式通过超声波辐射区，两支超声换能器交替工作，超声交替工作时间1~5s，提取时间30~80min； (2) Under the standard condition of ultrasonic power per liter of volume 200W, alternate dual-frequency countercurrent ultrasonic extraction is used, and the focused ultrasonic probes with frequencies of 15 kHz and 20 kHz are installed at a certain angle to each other, and the feed liquid flows from bottom to top. The method of countercurrent circulation passes through the ultrasonic radiation area, and the two ultrasonic transducers work alternately. The alternating working time of ultrasonic is 1~5s, and the extraction time is 30~80min;
（3）提取结束后，离心，将上清液浓缩、喷雾干燥，得到紫菜蛋白与多糖混合产品。 (3) After the extraction, centrifuge, concentrate and spray-dry the supernatant to obtain the mixed product of laver protein and polysaccharide.
本发明所述得到的一种紫菜蛋白与多糖混合产品，含有蛋白质15.50%～24.18%，多糖18.21%～24.00%。 The laver protein and polysaccharide mixed product obtained in the present invention contains 15.50%-24.18% of protein and 18.21%-24.00% of polysaccharide.
本发明的优点： Advantages of the present invention:
采用交替双频逆流超声辅助提取技术对条斑紫菜中多糖和蛋白进行同步提取，较常规提取方法提取效率更高；较单频超声辅助提取方法破壁和传质效果更强，能量利用率更高，更节能，更好地达到缩短提取时间和提高提取得率的目的，为高附加值紫菜蛋白与多糖混合产品的工业化生产提供一条可行之路。 Using alternating double-frequency countercurrent ultrasonic-assisted extraction technology to simultaneously extract polysaccharides and proteins from Porphyra zebra, the extraction efficiency is higher than conventional extraction methods; compared with single-frequency ultrasonic-assisted extraction methods, the effect of wall breaking and mass transfer is stronger, and the energy utilization rate is higher. Higher, more energy-saving, better achieve the purpose of shortening the extraction time and increasing the extraction yield, and provide a feasible way for the industrial production of high value-added laver protein and polysaccharide mixed products. the
附图说明 Description of drawings
图1是双频逆流超声辅助提取设备结构图，1为超声控制器，2为超声探头，3为盛液器，4为循环泵。 Figure 1 is a structural diagram of dual-frequency countercurrent ultrasonic-assisted extraction equipment, 1 is an ultrasonic controller, 2 is an ultrasonic probe, 3 is a liquid container, and 4 is a circulation pump.
具体实施方式 Detailed ways
本发明中以蛋白和多糖的总得率来评价产品的提取得率，总得率定义为粗蛋白中纯蛋白质量与原料质量的比值加上粗多糖中纯多糖质量与原料质量的比值。粗蛋白中蛋白含量的测定采用福林酚比色法；粗多糖中多糖含量的测定采用苯酚硫酸比色法。 In the present invention, the total yield of protein and polysaccharide is used to evaluate the extraction yield of the product, and the total yield is defined as the ratio of the pure protein amount in the crude protein to the raw material mass plus the ratio of the pure polysaccharide mass in the crude polysaccharide to the raw material mass. The determination of protein content in crude protein was carried out by Folin phenol colorimetric method; the determination of polysaccharide content in crude polysaccharide was carried out by phenol-sulfuric acid colorimetric method.
图1为本发明的交替双频逆流超声辅助水提法采用的双频逆流超声辅助提取设备，该设备配有超声控制器1，可控制两支超声探头2（f1和f2）分别发出频率为15kHz和20kHz的超声波；其中超声探头2（f1）和超声探头2（f2）相互呈一定角度（30~60度）安装；超声探头2插入盛液器3中进行超声辅助提取，启动循环泵4对提取液进行逆流循环。通过超声控制器1号控制超声探头工作功率、超声交替工作时间和提取总时间。
Figure 1 shows the dual-frequency countercurrent ultrasonic-assisted extraction equipment used in the alternate dual-frequency countercurrent ultrasonic-assisted water extraction method of the present invention. The equipment is equipped with an
the
对照例1：常规提取Comparative Example 1: Conventional Extraction
在交替双频逆流超声辅助提取的最优工艺下，采用不加超声的常规水提法进行紫菜蛋白和多糖的提取试验。将紫菜片粉碎至40目，取60g紫菜粉，按料液比14mg/mL加4.3L自来水溶解，加热至41℃，调节pH至9.0，浸提88min，5000rpm离心10min，取上清液，浓缩，将其喷雾干燥，得到紫菜蛋白与多糖混合产品，测得紫菜蛋白和多糖的总得率为17.95%，其中蛋白和多糖的得率分别为7.19%和10.76%。 Under the optimal process of alternating double-frequency countercurrent ultrasonic-assisted extraction, the extraction experiment of Porphyra protein and polysaccharide was carried out by conventional water extraction method without ultrasonic. Crush laver slices to 40 mesh, take 60g laver powder, add 4.3L tap water to dissolve according to the ratio of material to liquid 14mg/mL, heat to 41°C, adjust the pH to 9.0, extract for 88min, centrifuge at 5000rpm for 10min, take the supernatant, and concentrate , and spray-dried it to obtain a mixed product of laver protein and polysaccharide. The total yield of laver protein and polysaccharide was measured to be 17.95%, among which the yield of protein and polysaccharide was 7.19% and 10.76%, respectively.
对照例2：单频逆流超声辅助提取Control Example 2: Single-frequency Countercurrent Ultrasonic Assisted Extraction
在单频逆流超声辅助提取的最优工艺下，以与交替双频逆流超声辅助提取具有相同单位体积料液超声功率（200W/L）为比较基准，采用单一频率超声进行紫菜蛋白和多糖的提取试验。将紫菜片粉碎至40目，取20g紫菜粉，按料液比19mg/mL加1.1L自来水溶解，加热至40℃，调节pH至9.0，进行紫菜蛋白和多糖的单频逆流超声辅助提取。单频逆流超声辅助提取条件为：超声频率20kHz、超声脉冲工作时间4s、超声脉冲间隔时间2 s、提取时间85min，料液以逆流循环的方式通过超声探头。提取结束后，5000rpm离心10min，取上清液，浓缩，将其喷雾干燥，得到紫菜蛋白与多糖混合产品，测得紫菜蛋白和多糖的总得率为32.14%，其中蛋白和多糖的得率分别为13.42%和18.72%。 Under the optimal process of single-frequency countercurrent ultrasonic-assisted extraction, with the same ultrasonic power per unit volume (200W/L) as compared with alternate dual-frequency countercurrent ultrasonic-assisted extraction, a single-frequency ultrasonic was used for the extraction of laver protein and polysaccharides test. Crush laver flakes to 40 mesh, take 20 g of laver powder, dissolve in 1.1 L of tap water at a material-to-liquid ratio of 19 mg/mL, heat to 40 °C, adjust the pH to 9.0, and conduct single-frequency countercurrent ultrasonic-assisted extraction of laver proteins and polysaccharides. The single-frequency countercurrent ultrasonic-assisted extraction conditions are as follows: ultrasonic frequency 20 kHz, ultrasonic pulse working time 4 s, ultrasonic pulse interval time 2 s, extraction time 85 min, and the feed liquid passes through the ultrasonic probe in a countercurrent circulation manner. After the extraction, centrifuge at 5000rpm for 10min, take the supernatant, concentrate, and spray-dry it to obtain a mixed product of laver protein and polysaccharide. The total yield of laver protein and polysaccharide is 32.14%, wherein the yield of protein and polysaccharide is respectively 13.42% and 18.72%.
实施例1：Example 1:
在每升体积超声功率为200W的基准条件下，采用两个频率超声进行紫菜蛋白和多糖的提取试验。将紫菜片粉碎至40目，取60g紫菜粉，按料液比10mg/mL加6L自来水溶解，加热至80℃，调节pH至9.0，进行紫菜蛋白和多糖的交替双频逆流超声辅助提取。交替双频逆流超声辅助提取条件为：15 kHz和20 kHz双频交替工作、超声交替工作时间3 s、提取时间30 min，料液以逆流循环的方式通过超声探头。提取结束后，5000rpm离心10min，取上清液，浓缩，将其喷雾干燥，得到紫菜蛋白与多糖混合产品，测得紫菜蛋白和多糖的总得率为40.15%，其中蛋白和多糖的得率分别为21.02%和19.13%。 Under the reference condition of ultrasonic power of 200W per liter volume, the extraction experiment of Porphyra protein and polysaccharide was carried out by using two frequency ultrasonic waves. Crush laver flakes to 40 mesh, take 60 g of laver powder, dissolve in 6 L of tap water at a material-to-liquid ratio of 10 mg/mL, heat to 80 °C, adjust the pH to 9.0, and perform alternate dual-frequency countercurrent ultrasonic-assisted extraction of laver protein and polysaccharides. Alternate dual-frequency countercurrent ultrasonic-assisted extraction conditions are: 15 kHz and 20 kHz dual-frequency alternate operation, ultrasonic alternate working time 3 s, extraction time 30 min, and the feed liquid passes through the ultrasonic probe in a countercurrent circulation manner. After the extraction, centrifuge at 5000rpm for 10min, take the supernatant, concentrate, and spray-dry it to obtain the mixed product of laver protein and polysaccharide. The total yield of laver protein and polysaccharide is 40.15%, wherein the yield of protein and polysaccharide is respectively 21.02% and 19.13%.
实施例2：Example 2:
试验处理过程同实施例1，其不同在于其中料液比为15mg/mL，温度为80℃，提取时间为40min，紫菜蛋白和多糖的总得率为48.18%，其中蛋白和多糖的得率分别为24.18%和24.00%。 The test process is the same as in Example 1, the difference is that the solid-liquid ratio is 15mg/mL, the temperature is 80°C, the extraction time is 40min, the total yield of laver protein and polysaccharide is 48.18%, and the yield of protein and polysaccharide is respectively 24.18% and 24.00%.
实施例3：Example 3:
试验处理过程同实施例1，其不同在于其中料液比为20mg/mL，温度为80℃，提取时间为50min，紫菜蛋白和多糖的总得率为38.02%，其中蛋白和多糖的得率分别为15.50%和22.52%。 The test process is the same as in Example 1, the difference is that the solid-liquid ratio is 20mg/mL, the temperature is 80°C, the extraction time is 50min, the total yield of laver protein and polysaccharide is 38.02%, and the yield of protein and polysaccharide is respectively 15.50% and 22.52%.
实施例4：Example 4:
试验处理过程同实施例1，其不同在于其中温度为60℃，提取时间为40min，紫菜蛋白和多糖的总得率为37.56%，其中蛋白和多糖的得率分别为19.35%和18.21%。 The experimental treatment process was the same as in Example 1, except that the temperature was 60°C, the extraction time was 40 minutes, and the total yield of laver protein and polysaccharide was 37.56%, of which the yield of protein and polysaccharide was 19.35% and 18.21%, respectively.
实施例5：Example 5:
试验处理过程同实施例1，其不同在于其中温度为100℃，提取时间为40min，紫菜蛋白和多糖的总得率为45.15%，其中蛋白和多糖的得率分别为22.99%和22.16%。 The experimental treatment process was the same as in Example 1, the difference being that the temperature was 100°C, the extraction time was 40 minutes, the total yield of laver protein and polysaccharide was 45.15%, and the yield of protein and polysaccharide was 22.99% and 22.16%, respectively.
实施例6：Embodiment 6:
试验处理过程同实施例1，其不同在于其中提取时间为50min，紫菜蛋白和多糖的总得率为43.10%，其中蛋白和多糖的得率分别为22.58%和20.52%。 The experimental treatment process was the same as in Example 1, the difference being that the extraction time was 50 minutes, the total yield of laver protein and polysaccharide was 43.10%, and the yield of protein and polysaccharide was 22.58% and 20.52% respectively.
实施例7：Embodiment 7:
试验处理过程同实施例1，其不同在于其中温度为40℃，提取时间为80min，料液比15mg/mL，超声交替工作时间1s，紫菜蛋白和多糖的总得率为46.19%，其中蛋白和多糖的得率分别为23.32%和22.87%。 The experimental treatment process is the same as in Example 1, the difference is that the temperature is 40°C, the extraction time is 80min, the solid-liquid ratio is 15mg/mL, and the ultrasonic alternating working time is 1s. The total yield of laver protein and polysaccharide is 46.19%. The yields were 23.32% and 22.87%, respectively.
实施例8：Embodiment 8:
试验处理过程同实施例1，其不同在于其中温度为40℃，提取时间为80min，料液比15mg/mL，超声交替工作时间5s，紫菜蛋白和多糖的总得率为47.68%，其中蛋白和多糖的得率分别为24.04%和23.64%。 The experimental treatment process is the same as in Example 1, the difference is that the temperature is 40°C, the extraction time is 80min, the solid-liquid ratio is 15mg/mL, and the ultrasonic alternating working time is 5s. The total yield of laver protein and polysaccharide is 47.68%. The yields were 24.04% and 23.64%, respectively.
实施例9：Embodiment 9:
试验处理过程同实施例1，其不同在于其中温度为40℃，提取时间为60min，料液比20mg/mL，提取pH8.0，紫菜蛋白和多糖的总得率为35.59%，其中蛋白和多糖的得率分别为15.58%和20.01%。 The experimental treatment process is the same as in Example 1, the difference is that the temperature is 40°C, the extraction time is 60min, the solid-liquid ratio is 20mg/mL, the extraction pH is 8.0, and the total yield of laver protein and polysaccharide is 35.59%. The yields were 15.58% and 20.01%, respectively.
实施例10：Example 10:
试验处理过程同实施例1，其不同在于其中温度为40℃，提取时间为60min，料液比20mg/mL，提取pH10.0，紫菜蛋白和多糖的总得率为36.25%，其中蛋白和多糖的得率分别为15.68%和20.57%。 The test process is the same as in Example 1, the difference is that the temperature is 40°C, the extraction time is 60min, the solid-liquid ratio is 20mg/mL, the extraction pH is 10.0, and the total yield of laver protein and polysaccharide is 36.25%. The yields were 15.68% and 20.57%, respectively.
表1 不同实施例紫菜蛋白和多糖总得率 Table 1 Total yield of laver protein and polysaccharide in different examples
由表1可以看出交替双频逆流超声辅助提取与常规提取和单频逆流超声辅助提取相比，可以显著提高紫菜蛋白和多糖的提取得率。 It can be seen from Table 1 that, compared with conventional extraction and single-frequency countercurrent ultrasonic-assisted extraction, alternate dual-frequency countercurrent ultrasonic-assisted extraction can significantly improve the extraction yield of Porphyra protein and polysaccharides.
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