WO2023030023A1 - 一种利用酵母菌发酵制备多功能水凝胶的方法 - Google Patents

一种利用酵母菌发酵制备多功能水凝胶的方法 Download PDF

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WO2023030023A1
WO2023030023A1 PCT/CN2022/113331 CN2022113331W WO2023030023A1 WO 2023030023 A1 WO2023030023 A1 WO 2023030023A1 CN 2022113331 W CN2022113331 W CN 2022113331W WO 2023030023 A1 WO2023030023 A1 WO 2023030023A1
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pca
yeast
solution
gel
hydrogel
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汪少芸
程静
游力军
熊蔡华
蔡茜茜
陈选
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福州大学
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    • A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
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    • A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
    • A61L26/0009—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form containing macromolecular materials
    • A61L26/0023—Polysaccharides
    • A—HUMAN NECESSITIES
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  • the invention belongs to the field of preparation of conductive hydrogel, and in particular relates to a method for preparing multifunctional hydrogel by yeast fermentation.
  • Hydrogel is a kind of material with a three-dimensional polymer or supramolecular polymer network structure. It has good flexibility and can be pulled, pressed, bent, etc.
  • conductive hydrogel is a kind of material that can be added by adding conductive polymers.
  • Hydrogels prepared by physical crosslinking Their unique properties (such as flexibility, high water content, biocompatibility, electrical conductivity, etc.) make them widely used in various biomedical fields, including the detection of human physiological signals, regenerative medicine, neural repair, etc.
  • the poor gas permeability, mechanical properties, and water retention of conductive hydrogels greatly limit the application of conductive hydrogels in the field of biomedical materials. Therefore, it is of great significance to improve the air permeability, mechanical properties and water retention of hydrogels.
  • Biosensor is an important device for detecting and tracking human physiological signals
  • hydrogel is a new type of biosensor.
  • ECG ECG
  • EMG EEG
  • nerve signals it has high comfort, shape controllability and sensitivity.
  • Most of the current hydrogels have problems such as airtightness and poor mechanical strength. Therefore, the preparation A porous, breathable and high-strength conductive hydrogel has broad development and application prospects.
  • the preparation of conductive hydrogels currently reported contains bioincompatible synthetic polymers, toxic cross-linking agents, and complicated operating procedures. Therefore, it is important to develop a simple, fast, safe, efficient, and breathable method. significance.
  • the purpose of the present invention is to provide a method for preparing multifunctional hydrogel by fermentation of yeast to address the deficiencies in research in the above fields.
  • the method is simple, fast and efficient, and the obtained hydrogel has good air permeability, water retention, flexibility and conductivity.
  • a method for preparing multifunctional hydraulic gel by yeast fermentation comprising the following steps:
  • PCA Weigh Plate Count Agar
  • step (3) Stir the mixed solution of the yeast liquid in step (1) and the Gel-PCA-glucose in step (2), pour it into the mold, and put it in a 30°C water bath to ferment for 30min, then put it in The Gel-PCA-yeast multifunctional hydrogel was obtained at 4°C for 10 minutes.
  • the yeast liquid concentration described in the above step (1) is 0.2g/mL-0.45g/mL.
  • the concentration of the PCA solution in the above step (2) is 0.0235g/mL; the amount of gelatin added is 5wt%-35wt%; the amount of glucose added is 0.01-0.06g/mL.
  • step (2) add reduced graphene oxide (PrGO) with a concentration of 1-4 mg/mL to the PCA solution; obtain a Gel-PrGO-PCA-glucose mixed solution; Stir with the Gel-PrGO-PCA-glucose mixed solution, pour it into the mold, put it in a water bath at 30°C for 30 minutes, and then place it at 4°C for 10 minutes to obtain Gel-PrGO-PCA-yeast multifunctional water gel.
  • PrGO reduced graphene oxide
  • the preparation method of described reduced graphene oxide (PrGO) comprises:
  • the Gel-PCA-yeast hydrogel obtained in the above step (3) was frozen at -80°C for 20 minutes, cut into 1mm thick slices or cut into arbitrary shapes, and soaked in a mixed solution of saline solution and glycerol for 12 hours .
  • the salt solution is ammonium sulfate or sodium citrate solution; the concentration of the salt solution is 10wt%-30wt%; the volume ratio of the salt solution to glycerin is 2:1, 1:1 or 1:2.
  • the resulting multifunctional hydrogel prepared by any one of the above methods.
  • the above-mentioned multifunctional hydrogels are used as conductive materials in biosensors, or in drug-loaded and antibacterial wound dressings.
  • the present invention has the following advantages:
  • the present invention uses gelatin mixed with reduced graphene oxide, PCA, saccharomyces, and glucose to prepare by fermentation, and then soak it in a salt solution or a mixed solution of salt solution and glycerin.
  • the multifunctional hydrogel prepared by yeast fermentation has multiple functions, in which yeast endows it with porous air permeability, gelatin, PrGO and salt solution endow it with electrical conductivity and mechanical properties.
  • the hydrogel has anti-fatigue performance and super stretchability, and its stretchability can reach 1000%, and it can sensitively detect ECG and EMG signals, and is expected to be applied to biosensors or wearable devices, etc. field.
  • FIG. 1 Hydrogel morphology.
  • a Topography of hydrogel without yeast added under ordinary camera
  • b Topography of hydrogel without yeast added under optical microscope
  • c Topography of hydrogel with yeast added under ordinary camera Figure
  • d Topography of hydrogel with yeast added under optical microscope.
  • Fig. 3 Tensile stress-strain curves of Gel-PCA-yeast-ammonium sulfate hydrogel and Gel-PCA-yeast-sodium citrate hydrogel soaked in different salt solutions.
  • Figure 5 The morphology, electrical conductivity and detection of ECG and EMG signals of Gel-PrGO-PCA-yeast hydrogel.
  • a Morphology of Gel-PrGO-PCA-yeast hydrogel;
  • b conductivity detection;
  • c detection of ECG signal;
  • d detection of EMG signal.
  • Figure 6 Water retention properties of Gel-PCA-yeast and Gel-PCA-yeast-ammonium sulfate-glycerol hydrogels.
  • Figure 7 Rheological properties of Gel-PCA-yeast and Gel-PCA-yeast-ammonium sulfate-glycerol hydrogels.
  • the Gel-PCA-yeast-ammonium sulfate hydrogel and Gel-PCA-yeast-sodium citrate hydrogel obtained in the 20% saline solution soaking group have super tensile properties, wherein Gel -PCA-yeast-sodium citrate hydrogel has a maximum tensile strain of 1000% and a maximum tensile stress of 0.28MPa; Gel-PCA-yeast-ammonium sulfate hydrogel has a maximum tensile strain of 850% and a maximum tensile stress of 0.28MPa The tensile stress is 0.14MPa.
  • the Young's modulus of the ammonium sulfate group is smaller than that of the sodium citrate group ( Figure 4), indicating that it is more flexible or elastic. good.
  • the mechanical properties of the hydrogels soaked in 10% saline solution were poor, and the tensile properties were less than 100%.
  • a method for preparing multifunctional hydrogel by yeast fermentation comprising the following steps:
  • the obtained Gel-PrGO-PCA-yeast hydrogel has good conductivity, its conductivity is 0.015S/m, and it has a good network porous structure, which can detect heart well. electrical and myoelectric signals.
  • Rheological test results show that the storage modulus increases from 100Pa to 1000Pa when the ratio of ammonium sulfate and glycerol is 1:1, which is 2 orders of magnitude higher than that of the unsoaked group, indicating that the hydrogel strength after soaking is higher than that of the unsoaked group.

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Abstract

本发明涉及一种利用酵母菌发酵制备多功能水凝胶的方法。本发明首先利用聚多巴胺还原氧化石墨烯得到还原氧化石墨烯溶液，然后配制一定浓度的明胶PCA葡萄糖混合溶液以及活化的酵母菌溶液；通过一锅反应法将三者混合，搅拌均匀，倒入模具中，在30℃水浴中发酵一定的时间，即得Gel-PrGO-PCA-酵母多功能水凝胶材料。该方法简单方便、快速高效，所得水凝胶具有良好的透气性、超强的机械性能、导电性、生物相容性，并且可通过贴敷于不同的皮肤位置检测心电和肌电。本发明为制备多孔透气导电水凝胶电极提供一种新的思路及方法，有助于导电水凝胶材料的开发利用，以便其应用在生物传感器领域。

Description

一种利用酵母菌发酵制备多功能水凝胶的方法 技术领域
本发明属于导电水凝胶的制备领域，具体涉及一种利用酵母菌发酵制备多功能水凝胶的方法。
背景技术
水凝胶是一类具有三维聚合物或超分子聚合物网络结构的材料，具有很好的柔性，可以进行拉、压、弯曲等，其中导电水凝胶是一种通过添加导电高分子聚合物进行物理交联制备的水凝胶。它们独特的性质(如柔韧性、高含水量、生物相容性、导电性等)促使它们广泛应用于各种生物医学领域，包括人体生理信号的检测、再生医学、神经修复等领域。然而，导电水凝胶的透气性、机械性以及保水性差，大大限制了导电水凝胶在生物医用材料领域的应用。因此提高水凝胶的透气性、机械性能以及保水性具有重要意义。
技术问题
生物传感器是一种重要的检测和追踪人体生理信号的设备，其中水凝胶是一种新型的生物传感器。对于检测和追踪人体的心电、肌电、脑电以及神经信号具有较高的舒适性、形状可控性和灵敏性，目前的水凝胶大多存在不透气、机械强度差等问题，因此制备一种多孔透气高强度的导电水凝胶具有广大的发展和应用前景。并且目前有报道的导电水凝胶的制备含有生物不相容的合成高分子、有毒的交联剂、复杂的操作流程，因此，开发一种简单、快速、安全、高效、透气的方法具有重要意义。
技术解决方案
本发明的目的在于针对上述领域研究的不足，提供一种利用酵母菌发酵制备多功能水凝胶的方法。该方法操作简单、快速、高效，且所得水凝胶具有良好的透气性、保水性、柔韧性和导电性。
为实现上述目的，采用以下技术方案：
一种利用酵母菌发酵制备多功能水凝的方法，包括如下步骤：
(1)将酵母菌在30℃的热水中活化得到酵母菌液；
(2)称取平板计数琼脂(PCA)溶解在100℃去离子水中煮沸20min，冷却至50℃，得PCA溶液；向PCA溶液中加入明胶(Gel)于50℃水浴中搅拌30min，然后加入葡萄糖，继续搅拌溶解得到Gel-PCA-葡萄糖的混合溶液，降温至30℃；
(3)将步骤(1)的酵母菌液和步骤(2)的Gel-PCA-葡萄糖的混合溶液搅拌混匀后倒入模具中，并置于30℃水浴中发酵30min，然后将其放在4℃下10min即得到Gel-PCA-酵母菌多功能水凝胶。
上述步骤(1)中所述酵母菌液浓度为0.2g/mL-0.45g/mL。
上述步骤(2)中所述PCA溶液的浓度为0.0235g/mL；明胶的添加量为5wt％-35wt％；所述的葡萄糖的添加量为0.01-0.06g/mL。
进一步的，上述步骤(2)中向PCA溶液中加入浓度为1-4mg/mL的还原氧化石墨烯(PrGO)；得到Gel-PrGO-PCA-葡萄糖混合溶液；将步骤(1)的酵母菌液和Gel-PrGO-PCA-葡萄糖混合溶液搅拌混匀后倒入模具中，并置于30℃水浴中发酵30min，然后将其放在4℃下10min即得到Gel-PrGO-PCA-酵母多功能水凝胶。
更进一步的，所述还原氧化石墨烯(PrGO)的制备方法包括：
(1)氧化石墨烯(GO)的制备：称取1.2g石墨，加入50mL浓硫酸，搅拌均匀后，放入冰浴中，搅拌状态下加入1.5g硝酸钠，并缓慢加入6g高锰酸钾，于35℃下搅拌过夜，缓慢加入去离子水100mL，控制温度为90℃，反应1h，将30vol％的过氧化氢稀释5倍后缓慢加入上述溶液中，直到不产生气泡，停止添加，继续反应3h，冷却至室温，水洗至中性后超声分散20min，冻干后得到氧化石墨烯(GO)粉末；
(2)称取20mg GO粉末，加入2.5mL去离子水，超声直至完全溶解，得到GO分散液；称取50mg盐酸多巴胺(DA)粉末溶解在2.5mL 10mM Tris-HCl溶液(pH＝8.5)中，得盐酸多巴胺分散液；然后将盐酸多巴胺分散液加入到GO分散液中，冰浴下超声分散2h后，在60℃水浴下搅拌12h，得到还原氧化石墨烯PrGO溶液。
进一步的，上述步骤(3)中所得Gel-PCA-酵母菌水凝胶放在-80℃冷冻20min后切成1mm厚的薄片或切成任意形状，浸泡在盐溶液和甘油的混合溶液中12h。 所述盐溶液为硫酸铵或柠檬酸钠溶液；所述的盐溶液浓度为10wt％-30wt％；所述盐溶液与甘油的体积比为2：1、1：1或1：2。
上述任一项方法制备所得的多功能水凝胶。
上述多功能水凝胶作为导电材料用于生物传感器中，或用于载药、抗菌伤口敷料中。
有益效果
相对于现有技术，本发明具有以下优点：
(1)本发明采用明胶与还原氧化石墨烯、PCA、酵母菌、葡萄糖混合后进行发酵制备，然后将其浸泡在盐溶液或盐溶液与甘油的混合溶液中。
(2)该利用酵母菌发酵制备的多功能水凝胶具有多种功能，其中酵母菌赋予了其多孔透气性，明胶、PrGO和盐溶液赋予了其导电性和机械性能。
(3)该水凝胶具有抗疲劳性能和超强的拉伸性能，其拉伸性能可达到1000％，并且能灵敏的检测心电和肌电信号，有望应用于生物传感器或可穿戴设备等领域。
附图说明
图1添加不同含量的明胶对孔径的影响。
图2水凝胶形貌图。a，普通相机下未添加酵母菌的水凝胶的形貌图；b，光学显微镜下未添加酵母菌的水凝胶形貌图；c，普通相机下添加酵母菌的水凝胶的形貌图；d，光学显微镜下添加酵母菌的水凝胶形貌图。
图3 Gel-PCA-酵母菌-硫酸铵水凝胶和Gel-PCA-酵母菌-柠檬酸钠水凝胶浸泡不同盐溶液后的拉伸应力-应变曲线。
图4杨氏模量图。
图5 Gel-PrGO-PCA-酵母菌水凝胶的形貌、导电性以及对心电和肌电信号的检测图。a，Gel-PrGO-PCA-酵母菌水凝胶的形貌；b，导电性检测；c，心电信号的检测；d，肌电信号的检测。
图6 Gel-PCA-酵母菌和Gel-PCA-酵母菌-硫酸铵-甘油水凝胶的保水性能。
图7 Gel-PCA-酵母菌和Gel-PCA-酵母菌-硫酸铵-甘油水凝胶的流变性能。
本发明的实施方式
以下结合实施实例对本发明做进一步说明，需要指出的是，本实施实例仅用于解释本发明，而非对本发明的限制。
实施例1
称取7份0.235g平板计数琼脂(PCA)分别溶解在9mL 100℃去离子水中煮沸20min，冷却至50℃，分别加入5wt％、10wt％、15wt％、20wt％、25wt％、30wt％和35wt％明胶(Gel)于50℃水浴中搅拌30min，然后加入0.15g葡萄糖，继续搅拌溶解得到Gel-PCA和葡萄糖的混合溶液，降温至30℃；称取0.45g酵母粉溶解于1mL 30℃去离子水中，然后将酵母菌液和上述Gel-PCA-葡萄糖的混合溶液搅拌混匀后倒入模具中，并置于30℃水浴中发酵30min，然后将其放在4℃下10min即得到Gel-PCA-酵母菌水凝胶。最后将其放在-80℃冷冻20min后切成1mm厚的薄片或切成任意形状，浸泡在20wt％的硫酸铵和甘油(体积比1：1)的混合溶液中12h后进行检测。如图1所示，得到的Gel-PCA-酵母菌水凝胶随着明胶含量的增加，水凝胶的孔径先减小后增加。
实施例2
将0.235g PCA溶解在9mL 100℃去离子水中煮沸20min，冷却至50℃，加入20wt％明胶于50℃水浴中搅拌30min，然后加入0.15g葡萄糖，继续搅拌溶解得到Gel-PCA和葡萄糖的混合溶液，降温至30℃；称取0.45g酵母粉溶解于1mL 30℃去离子水中(对照组不添加酵母菌)，然后将酵母菌液和上述Gel-PCA-葡萄糖的混合溶液搅拌混匀后倒入模具中，并置于30℃水浴中发酵30min，然后将其放在4℃下10min即得到Gel-PCA-酵母菌水凝胶。最后将其放在-80℃冷冻20min后切成1mm厚的薄片或切成任意形状进行检测。如图2所示，得到的Gel-PCA-酵母菌水凝胶具有较好的网络多孔结构。
实施例3
将0.235g PCA溶解在9mL 100℃去离子水中煮沸20min，冷却至50℃，加入20wt％明胶于50℃水浴中搅拌30min，然后加入0.15g葡萄糖，继续搅拌溶解得到Gel-PCA和葡萄糖的混合溶液，降温至40℃；称取0.45g酵母粉溶解于1mL 30℃去离子水中；然后将酵母菌液和上述Gel-PCA和葡萄糖的混合溶液搅拌混匀后倒入模具中，并置于30℃水浴中发酵30min，然后将其放在4℃下10min即得到Gel-PCA-酵母菌水凝胶。切成所需要的形状后，将其分别浸泡到10wt％、20wt％和30wt％的硫酸铵或者柠檬酸钠的盐溶液中12h后，进行拉 伸性能检测。如图3所示，20％盐溶液浸泡组得到的Gel-PCA-酵母菌-硫酸铵水凝胶和Gel-PCA-酵母菌-柠檬酸钠水凝胶具有超强的拉伸性能，其中Gel-PCA-酵母菌-柠檬酸钠水凝胶最大拉伸应变达到1000％，最大拉伸应力为0.28MPa；Gel-PCA-酵母菌-硫酸铵水凝胶最大拉伸应变达到850％，最大拉伸应力为0.14MPa，虽然柠檬酸钠组的拉伸性能大于硫酸铵组，但是硫酸铵组的杨氏模量小于柠檬酸钠组的杨氏模量(图4)，表明其柔性或弹性较好。10％盐溶液浸泡组的水凝胶的力学性能较差，拉伸性能小于100％。
实施例4
一种利用酵母菌发酵制备多功能水凝胶的方法，包括以下步骤：
(1)氧化石墨烯(GO)的制备：称取1.2g石墨，加入50mL浓硫酸，搅拌均匀后，放入冰浴中，搅拌状态下加入1.5g硝酸钠，并缓慢加入6g高锰酸钾，于35℃下搅拌过夜，缓慢加入去离子水100mL，控制温度为90℃，反应1h，将30vol％的过氧化氢稀释5倍后缓慢加入上述溶液中，直到不产生气泡，停止添加，继续反应3h，冷却至室温，水洗至中性后超声分散20min，冻干后得到GO粉末。
(2)称取20mg GO粉末，加入2.5mL去离子水，超声直至完全溶解，得到GO分散液；称取50mg盐酸多巴胺(DA)粉末溶解在2.5mL 10mM Tris-HCl溶液(pH＝8.5)中，得盐酸多巴胺分散液；然后将盐酸多巴胺分散液加入到GO分散液中，冰浴下超声分散2h后，在60℃水浴下搅拌12h，得到还原氧化石墨烯PrGO溶液。
(3)将0.235g PCA溶解在4mL 100℃去离子水中，煮沸20min，冷却至50℃，与步骤(2)制备的PrGO溶液混匀后，加入20wt％明胶于50℃水浴中搅拌30min，然后加入0.15g的葡萄糖，继续搅拌溶解得到Gel-PrGO-PCA-葡萄糖的混合溶液，降温至30℃。称取0.45g酵母粉溶解于1mL 30℃去离子水中，得酵母菌液。然后将酵母菌液和上述Gel-PrGO-PCA-葡萄糖的混合溶液搅拌混匀后倒入模具中，并置于30℃水浴中发酵30min，然后将其放在4℃下10min即得到Gel-PrGO-PCA-酵母菌水凝胶。如图5所示，得到的Gel-PrGO-PCA-酵母菌水凝胶具有较好的导电性，其电导率为0.015S/m,且具有较好的网络多孔结构，能很好的检测心电和肌电信号。
实施例5
将0.235g PCA溶解在9mL 100℃去离子水中煮沸20min，冷却至50℃，加入20wt％明胶于50℃水浴中搅拌30min，然后加入0.15g的葡萄糖，继续搅拌溶解得到Gel-PCA和葡萄糖的混合溶液，降温至30℃。称取0.45g酵母粉溶解于1mL 30℃去离子水中。然后将酵母菌液和上述Gel-PCA和葡萄糖的混合溶液搅拌混匀后倒入模具中，并置于30℃水浴中发酵30min，然后将其放在4℃下10min即得到Gel-PCA-酵母菌水凝胶。最后将其浸泡在20wt％硫酸铵和甘油的混合溶液中(1：2，1：1，2：1(v/v))12h后，进行检测。如图6和图7所示，得到的Gel-PCA-酵母菌-硫酸铵-甘油水凝胶具有较好的保水性和流变性能，浸泡硫酸铵和甘油比例为1：1和1：2时，在室温下放置三天后其质量几乎不变，表明其失水较少。流变检测结果显示浸泡硫酸铵和甘油比例为1：1时其储能模量从100Pa增加至1000Pa，高出未浸泡组2个数量级，表明浸泡后的水凝胶强度高于未浸泡组。
以上所述仅为本发明的较佳实施例，凡依本发明申请专利范围所做的均等变化与修饰，皆应属本发明的涵盖范围。

Claims (9)

  1. 一种利用酵母菌发酵制备多功能水凝胶的方法，其特征在于，包括如下步骤：
    (1)将酵母菌在30℃的热水中活化得到酵母菌液；
    (2)称取平板计数琼脂PCA溶解在100℃去离子水中煮沸20min，冷却至50℃，得PCA溶液；向PCA溶液中加入明胶Gel于50℃水浴中搅拌30min，然后加入葡萄糖，继续搅拌溶解得到Gel-PCA-葡萄糖的混合溶液，降温至30℃；
    (3)将步骤(1)的酵母菌液和步骤(2)的Gel-PCA-葡萄糖的混合溶液搅拌混匀后倒入模具中，并置于30℃水浴中发酵30min，然后将其放在4℃下10min即得到Gel-PCA-酵母菌多功能水凝胶。
  2. 根据权利要求1所述的一种利用酵母菌发酵制备多功能水凝的方法，其特征在于：步骤(1)中所述酵母菌液浓度为0.2g/mL-0.45g/mL。
  3. 根据权利要求1所述的一种利用酵母菌发酵制备多功能水凝的方法，其特征在于：步骤(2)中所述PCA溶液的浓度为0.0235g/mL；明胶的添加量为5wt％-35wt％；所述的葡萄糖的添加量为0.01-0.06g/mL。
  4. 根据权利要求1所述的一种利用酵母菌发酵制备多功能水凝的方法，其特征在于：步骤(2)中向PCA溶液中加入浓度为1-4mg/mL的还原氧化石墨烯PrGO；得到Gel-PrGO-PCA-葡萄糖混合溶液；将步骤(1)的酵母菌液和Gel-PrGO-PCA-葡萄糖混合溶液搅拌混匀后倒入模具中，并置于30℃水浴中发酵30min，然后将其放在4℃下10min即得到Gel-PrGO-PCA-酵母菌多功能水凝胶。
  5. 根据权利要求4所述的一种利用酵母菌发酵制备多功能水凝的方法，其特征在于：所述还原氧化石墨烯的制备方法，包括以下步骤：
    (1)氧化石墨烯的制备：称取1.2g石墨，加入50mL浓硫酸，搅拌均匀后，放入冰浴中，搅拌状态下加入1.5g硝酸钠，并缓慢加入6g高锰酸钾，于35℃下搅拌过夜，缓慢加入去离子水100mL，控制温度为90℃，反应1h，将30vol％的过氧化氢稀释5倍后缓慢加入上述溶液中，直到不产生气泡，停止添加，继续反应3h，冷却至室温，水洗至中性后超声分散20min，冻干后得到氧化石墨烯粉末；
    (2)称取20mg GO粉末，加入2.5mL去离子水，超声直至完全溶解，得到GO分散液；称取50mg盐酸多巴胺粉末溶解在2.5mL 10mM pH＝8.5Tris-HCl溶液中，得盐酸多巴胺分散液；然后将盐酸多巴胺分散液加入到GO分 散液中，冰浴下超声分散2h后，在60℃水浴下搅拌12h，得到还原氧化石墨烯PrGO溶液。
  6. 根据权利要求1所述的一种利用酵母菌发酵制备多功能水凝的方法，其特征在于：步骤(3)中所得Gel-PCA-酵母菌水凝胶放在-80℃冷冻20min后切成1mm厚的薄片或切成任意形状，浸泡在盐溶液和甘油的混合溶液中12h。
  7. 根据权利要求6所述的一种利用酵母菌发酵制备多功能水凝的方法，其特征在于：所述盐溶液为硫酸铵或柠檬酸钠溶液；所述的盐溶液浓度为10wt％-30wt％；所述盐溶液与甘油的体积比为2：1、1：1或1：2。
  8. 如权利要求1-7任一项方法制备所得的多功能水凝胶。
  9. 如权利要求8所述多功能水凝胶的应用，其特征在于：多功能水凝胶作为导电材料用于生物传感器中，或用于载药、抗菌伤口敷料中。
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