CN109490274B - Experimental device for researching unidirectional mass transfer of enzyme in leather and application method - Google Patents
Experimental device for researching unidirectional mass transfer of enzyme in leather and application methodDownload PDFInfo
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Abstract
Translated fromChinese
Description
Translated fromChinese技术领域technical field
本发明属于制革酶制剂的应用研究领域，具体涉及一种用于研究酶在皮革中单向传质的实验装置及使用方法。The invention belongs to the applied research field of tanning enzyme preparations, and in particular relates to an experimental device and a use method for studying one-way mass transfer of enzymes in leather.
背景技术Background technique
制革行业作为畜牧业的延伸，是循环经济的重要环节，在我国工业经济中占有重要位置。随着国家对环境保护越来越重视，以生物酶为基础的清洁化制革技术得到迅速的发展和广泛的应用。与常见的酶促反应不同，制革生产中酶的催化底物——动物皮是一种由胶原纤维构成的具有三维立体结构的结缔组织。酶与动物皮作用的本质是酶分子进入皮纤维间隙并对动物皮不同组分进行水解，其可以分为传质和反应两个过程。研究表明酶的传质占整个反应时间的70%以上，因此传质是酶对动物皮有效作用的先决条件和限速步骤。研究酶在皮革中传质过程，实现对皮革不同深度酶浓度的准确定量，是酶制剂在制革工业推广和应用的理论基础。As an extension of animal husbandry, the tanning industry is an important part of the circular economy and occupies an important position in my country's industrial economy. As the country pays more and more attention to environmental protection, the clean tanning technology based on biological enzymes has been rapidly developed and widely used. Different from common enzymatic reactions, the catalytic substrate of enzymes in tanning production—animal skin is a connective tissue with a three-dimensional structure composed of collagen fibers. The essence of the interaction between enzymes and animal skins is that enzyme molecules enter the gap between skin fibers and hydrolyze different components of animal skins, which can be divided into two processes: mass transfer and reaction. Studies have shown that the mass transfer of enzymes accounts for more than 70% of the entire reaction time, so mass transfer is the prerequisite and rate-limiting step for the effective action of enzymes on animal skins. Studying the mass transfer process of enzymes in leather and realizing accurate quantification of enzyme concentrations at different depths of leather is the theoretical basis for the promotion and application of enzyme preparations in the leather industry.
目前，最常见的研究方法是利用荧光标记技术将酶或者蛋白标记后，再通过荧光显微镜或者激光扫描共聚焦荧光显微镜观察和记录皮革纵切片的荧光强度，获得酶在皮革中分布数据。但由于荧光标记酶或者荧光标记蛋白制备非常困难，而常规实验方法荧光标记酶或者荧光标记蛋白的用量大，因此难以在目前实验室常见的制革实验设备上进行传质研究。At present, the most common research method is to use fluorescent labeling technology to label enzymes or proteins, and then observe and record the fluorescence intensity of leather longitudinal sections through a fluorescence microscope or laser scanning confocal fluorescence microscope to obtain enzyme distribution data in leather. However, because the preparation of fluorescently labeled enzymes or fluorescently labeled proteins is very difficult, and conventional experimental methods use a large amount of fluorescently labeled enzymes or fluorescently labeled proteins, it is difficult to conduct mass transfer research on the common tanning experimental equipment in the laboratory.
酶在皮革生产的应用，以传质方式不同可分为单向传质方式（如采用涂脱毛糊法脱毛）和湍流传质方式(如浸水过程、软化过程、转鼓中的酶脱毛过程等)。本专利所实现的实验装置可以模拟酶在皮革中的单向传质过程。The application of enzymes in leather production can be divided into unidirectional mass transfer methods (such as depilation by applying hair removal paste) and turbulent mass transfer methods (such as immersion process, softening process, enzyme depilation process in drums, etc.) according to different mass transfer methods. ). The experimental device realized in this patent can simulate the one-way mass transfer process of enzyme in leather.
发明内容Contents of the invention
针对现有酶在皮革中传质研究技术和实验设备对样品需求量大、实验过程不易控制等问题，本发明提供了一种用于研究酶在皮革中单向传质的实验装置，实现在实验室条件下，利用极少量实验材料即可模拟酶在皮革中的单向传质过程。Aiming at the problems of the existing enzyme mass transfer research technology in leather and experimental equipment that require a large amount of samples and the experimental process is difficult to control, the present invention provides an experimental device for studying the one-way mass transfer of enzyme in leather, which can be realized in Under laboratory conditions, the one-way mass transfer process of enzymes in leather can be simulated with a very small amount of experimental materials.
本发明所要解决的技术问题通过以下的技术方案实现：一种用于研究酶在皮革中单向传质的实验装置，由上储液池、下储液池、底座、水平泡、可调节F夹、底座调节螺栓和下夹调节螺丝构成；The technical problem to be solved by the present invention is achieved through the following technical solutions: an experimental device for studying one-way mass transfer of enzymes in leather, which consists of an upper liquid storage tank, a lower liquid storage tank, a base, a horizontal bubble, an adjustable F Clamp, base adjustment bolt and lower clamp adjustment screw;
所述上储液池和下储液池均为一端开口的桶状结构，上储液池和下储液池口对口放置，皮革试样放置在在上、下储液池之间；The upper liquid storage tank and the lower liquid storage tank are barrel-shaped structures with one end open, the upper liquid storage tank and the lower liquid storage tank are placed mouth to mouth, and the leather sample is placed between the upper and lower liquid storage tanks;
所述底座底面四角安装有底座调节螺栓，上面安装有水平泡，通过调节水平泡的位置来确保储液池和皮革试样的水平；Base adjustment bolts are installed at the four corners of the bottom surface of the base, and a leveling bubble is installed on the top, and the level of the liquid storage tank and the leather sample is ensured by adjusting the position of the leveling bubble;
所述可调节F夹垂直的固定在底座上，可调节F夹下夹安装有下夹调节螺丝，可调节F夹下夹上下滑动用来粗调上下夹之间的距离，便于不同规格储液池的更换和实验过程中储液池的安装和拆卸；旋转下夹调节螺丝将皮革试样固定在上储液池和下储液之间。The adjustable F clamp is vertically fixed on the base, and the lower clamp of the adjustable F clamp is equipped with a lower clamp adjustment screw, which can be adjusted to slide up and down to roughly adjust the distance between the upper and lower clamps, which is convenient for different specifications of liquid storage Replacement of the pool and installation and removal of the liquid storage tank during the experiment; rotate the lower clip adjustment screw to fix the leather sample between the upper liquid storage tank and the lower storage liquid.
上储液池和下储液池材质、内径、外径及高度一致，可根据样品量的大小和实验的需求选择不同规格的储液池，实验过程中口对口将皮革试样固定在在上、下储液池之间，先将下储液池用医用注射器注满实验缓冲液，再将上储液池通过医用注射器注入一定量含有经过荧光标记的酶的溶液。The material, inner diameter, outer diameter and height of the upper liquid storage tank and the lower liquid storage tank are consistent. Different specifications of liquid storage tanks can be selected according to the size of the sample and the needs of the experiment. During the experiment, the leather sample is fixed on the upper surface Between the lower liquid storage tank and the lower liquid storage tank, first fill the lower liquid storage tank with the experimental buffer solution with a medical syringe, and then inject a certain amount of solution containing fluorescently labeled enzymes into the upper liquid storage tank through a medical syringe.
上储液池和下储液池的制作材料为具有弹性的硅胶，硅胶厚度为3~8mm。储液池的制作材料为具有弹性的硅胶，以便于通过注射器向储液池注射溶液或者从储液池抽取溶液样品，且在实验过程中可以将皮革试样牢固的固定在两储液池中间，而不产生漏液现象。储液池壁厚不低于3 mm以利于上下储液池与皮革样品的固定和医用注射器针眼的快速愈合；储液池壁厚不大于8 mm以利于医用注射器的刺入。The upper liquid storage tank and the lower liquid storage tank are made of elastic silica gel, and the thickness of the silica gel is 3-8mm. The reservoir is made of elastic silica gel, so that the solution can be injected into the reservoir or the solution sample can be drawn from the reservoir through a syringe, and the leather sample can be firmly fixed between the two reservoirs during the experiment , without leakage. The wall thickness of the reservoir is not less than 3 mm to facilitate the fixation of the upper and lower reservoirs and the leather sample and the rapid healing of the needle hole of the medical syringe; the wall thickness of the reservoir is not greater than 8 mm to facilitate the penetration of the medical syringe.
本发明中所述的一种模拟酶在皮革中单向传质的实验装置，其使用方法为：调节底座调节螺栓，使气泡位于水平泡的中心，取上储液池和下储液池各一个，下储液池装入80%容量的实验缓冲液，粒面向上将皮革试样遮盖于下储液池上，取上储液池口对口放在皮革试样上，保持与下储液池同心，调节可调节F夹下夹的位置，逆时针旋转下夹调节螺丝将上储液池、皮革试样及下储液池固定；先将下储液池通过医用注射器注满实验缓冲液，再将上储液池通过医用注射器注满含有荧光标记的酶溶液，传质结束后，取下皮革试样，冷冻切片后，观察和记录皮革切片荧光强度的分布情况。An experimental device for simulating one-way mass transfer of enzymes in leather described in the present invention is used as follows: adjust the base adjusting bolt so that the air bubbles are located in the center of the horizontal bubbles, and take each of the upper liquid storage tank and the lower liquid storage tank. One, fill the lower storage tank with 80% of the experimental buffer solution, cover the leather sample on the lower storage tank with the grain side up, take the upper storage tank and place it on the leather sample mouth to mouth, and keep it concentric with the lower storage tank , adjust the position of the lower clamp of the adjustable F clamp, and rotate the lower clamp adjustment screw counterclockwise to fix the upper reservoir, the leather sample and the lower reservoir; first fill the lower reservoir with the experimental buffer through a medical syringe, and then Fill the upper reservoir with the enzyme solution containing fluorescent markers through a medical syringe. After the mass transfer, remove the leather sample, freeze and section it, and observe and record the distribution of the fluorescence intensity of the leather section.
进一步地，所述的皮革试样为动物皮的任意部位。Further, the leather sample is any part of animal skin.
有益效果Beneficial effect
（1）在实验室条件下，利用极少量实验材料即可模拟酶在皮革中的单向传质过程，且可根据样品量的大小和实验的需求选择不同规格的储液池。(1) Under laboratory conditions, the one-way mass transfer process of enzymes in leather can be simulated with a very small amount of experimental materials, and different specifications of liquid storage tanks can be selected according to the size of the sample and the needs of the experiment.
（2）储液池的制作材料为具有弹性的硅胶，以便于通过注射器向储液池注射溶液或者从储液池抽取溶液样品，且在实验过程中可以将皮革试样牢固的固定在两储液池中间，而不产生漏液现象。(2) The material of the reservoir is made of elastic silica gel, so that the solution can be injected into the reservoir or the solution sample can be drawn from the reservoir through a syringe, and the leather sample can be firmly fixed in the two reservoirs during the experiment. In the middle of the liquid pool, without leakage.
（3）储液池壁厚不低于3 mm以利于上下储液池与皮革样品的固定和医用注射器针眼的快速愈合；储液池壁厚不大于8 mm以利于医用注射器的刺入。(3) The wall thickness of the reservoir is not less than 3 mm to facilitate the fixation of the upper and lower reservoirs and leather samples and the rapid healing of the needle hole of the medical syringe; the wall thickness of the reservoir is not greater than 8 mm to facilitate the penetration of the medical syringe.
（4）采用可调节F夹实现皮革试样的固定，F夹下夹在滑轨上的滑动可以粗调上下夹之间的距离，便于不同规格储液池的更换和实验过程中储液池的拆卸；通过旋紧下夹调节螺丝可以将皮革试样牢固的固定在两储液池之间。(4) The adjustable F clip is used to fix the leather sample. The sliding of the F clip on the slide rail can roughly adjust the distance between the upper and lower clips, which is convenient for the replacement of different specifications of the storage tank and the storage tank during the experiment. The disassembly of the leather sample can be firmly fixed between the two reservoirs by tightening the lower clamp adjustment screw.
附图说明Description of drawings
图1 研究酶在皮革中的单向传质的实验装置结构示意图；Figure 1 Schematic diagram of the experimental device for studying the one-way mass transfer of enzymes in leather;
其中，1. 上储液池；2.皮革试样；3.下储液池；4.底座；5.水平泡；6.可调节F夹；7.底座调节螺栓； 8.下夹调节螺丝；Among them, 1. Upper liquid storage tank; 2. Leather sample; 3. Lower liquid storage tank; 4. Base; 5. Level bubble; 6. Adjustable F clip; 7. Base adjustment bolt; ;
图2 FITC-Trypsin传质5min后皮革试样纵切片的荧光显微照片；Fig. 2 Fluorescence micrographs of longitudinal sections of leather samples after FITC-Trypsin mass transfer for 5 min;
图3 FITC-Trypsin传质30min后皮革试样纵切片的荧光显微照片；Figure 3 Fluorescence micrographs of longitudinal sections of leather samples after FITC-Trypsin mass transfer for 30 min;
图4 FITC-Trypsin传质30min后皮革试样不同深度横切片的荧光强度分布图；Figure 4. Fluorescence intensity distribution diagrams of cross-sections of leather samples at different depths after FITC-Trypsin mass transfer for 30 min;
图5 FITC-Dispase传质30min后皮革试样纵切片的荧光显微照片；Figure 5 Fluorescence micrographs of longitudinal sections of leather samples after FITC-Dispase mass transfer for 30 min;
图6 FITC-Dispase传质30min后皮革试样不同深度横切片的荧光强度分布图；Figure 6. Fluorescence intensity distribution diagrams of cross-sections of leather samples at different depths after FITC-Dispase mass transfer for 30 min;
图7 RBITC-Trypsin传质30min后皮革试样不同深度横切片的荧光强度分布图。Fig. 7 The fluorescence intensity distribution diagram of cross-sections of leather samples at different depths after RBITC-Trypsin mass transfer for 30 min.
具体实施方式Detailed ways
以下结合附图和具体实施例对本发明作进一步的详细描述，但本发明的保护范围并不仅限于下述实施例：Below in conjunction with accompanying drawing and specific embodiment the present invention is described in further detail, but protection scope of the present invention is not limited to following embodiment:
实施例1~6中传质实验使用的实验装置如图1所示，其中，1. 上储液池 2.皮革试样 3.下储液池 4.底座 5.水平泡 6.可调节F夹7.底座调节螺栓 8. 下夹调节螺丝；The experimental device used in the mass transfer experiment in Examples 1 to 6 is as shown in Figure 1, wherein, 1. upper
上储液池1和下储液池3均为一端开口的桶状结构，上储液池1和下储液池3口对口放置，皮革试样2放置在上储液池1和下储液池3之间；The upper
底座4的底面四角安装有底座调节螺栓7，上面安装有水平泡5；The four corners of the bottom surface of the
可调节F夹6垂直的固定在底座4上，可调节F夹6下夹上下滑动粗调上下夹之间的距离，可调节F夹6下夹安装有下夹调节螺丝8，旋转下夹调节螺丝8将皮革试样2牢牢固定在上储液池1和下储液池3之间。The
上储液池1和下储液池3的材质、内径、外径及高度完全一致，The material, inner diameter, outer diameter and height of the
上储液池1和下储液池3的制作材料为具有弹性的硅胶，硅胶厚度为3~8mm。The upper
下储液池3用于盛装实验缓冲液，上储液池1用于盛装含有经过荧光标记的酶的溶液。The
实施例1：Example 1:
（1）荧光标记酶的制备：取胰蛋白酶（Trypsin）、硫氰酸荧光素（Fluoresceinisothiocyanate，FITC）按照100: 1的质量比溶解于碳酸缓冲液中（0.05mol/L pH9.0），4℃下避光搅拌反应12~14h，反应完成后，冷冻干燥，将冻干粉重新溶解，利用凝胶层析（Sephadex G25填料）去除未反应的FITC，分离完后，测量FITC标记的胰蛋白酶溶液的荧光强度和浓度的关系。(1) Preparation of fluorescently labeled enzymes: Dissolve trypsin and fluorescein thiocyanate (FITC) in carbonic acid buffer (0.05mol/L pH9.0) at a mass ratio of 100:1, 4 Stir the reaction at ℃ for 12~14 hours in the dark. After the reaction is completed, lyophilize and redissolve the lyophilized powder. Use gel chromatography (Sephadex G25 filler) to remove unreacted FITC. After separation, measure FITC-labeled trypsin The relationship between the fluorescence intensity and the concentration of the solution.
（2）皮革试样裁取：山羊皮取脊背部，常规法脱毛，用圆形裁刀裁成直径50mm的皮革试样。(2) Cutting of leather samples: the back of the goatskin is taken, depilated by conventional methods, and cut into leather samples with a diameter of 50 mm with a circular cutter.
（3）传质实验：实验装置如图1所示，调节底座调节螺栓，使气泡位于水平泡的中心，取外径40mm的上储液池和下储液池各一个，下储液池装入80%容量的实验缓冲液（0.05mol/L pH7.4的磷酸缓冲液），粒面向上将皮革试样遮盖于下储液池上，将上储液池口对口放在皮革试样上，保持与下储液池同心，调节可调节F夹的下夹位置固定上、下储液池和皮革试样，逆时针旋转下夹调节螺丝将上、下储液池及皮革试样夹紧。先将下储液池通过医用注射器注满实验缓冲液，再将上储液池通过医用注射器注满含有FITC标记胰蛋白酶的溶液，传质5 min后，顺时针旋转下夹调节螺丝，取下皮革试样。(3) Mass transfer experiment: The experimental device is shown in Figure 1. Adjust the adjusting bolts of the base so that the air bubble is located in the center of the horizontal bubble. Take an upper liquid storage tank and a lower liquid storage tank with an outer diameter of 40mm, and install the lower liquid storage tank. Fill in 80% of the experimental buffer (0.05mol/L pH7.4 phosphate buffer), cover the leather sample on the lower reservoir with the grain side up, and place the upper reservoir mouth-to-mouth on the leather sample, keeping Concentric with the lower liquid storage tank, adjust the position of the lower clamp of the adjustable F clamp to fix the upper and lower liquid storage tanks and the leather sample, and turn the lower clamp adjustment screw counterclockwise to clamp the upper and lower liquid storage tanks and the leather sample. First fill the lower reservoir with the experimental buffer through a medical syringe, and then fill the upper reservoir with a solution containing FITC-labeled trypsin through a medical syringe. After mass transfer for 5 minutes, turn the lower clip adjustment screw clockwise to remove Leather samples.
（4）数据采集在皮革试样的中心取样，荧光显微镜观察皮革试样纵切片荧光强度分布情况。图2为FITC标记胰蛋白酶（FITC-Trypsin）传质5 min后皮革试样纵切片的荧光显微照片。(4) Data collection Sampling was done at the center of the leather sample, and the fluorescence intensity distribution of the longitudinal section of the leather sample was observed with a fluorescence microscope. Figure 2 is a fluorescence micrograph of a longitudinal section of a leather sample after mass transfer with FITC-Trypsin for 5 min.
实施例2：Example 2:
（1）荧光标记酶的制备：取胰蛋白酶（Trypsin）、硫氰酸荧光素（Fluoresceinisothiocyanate，FITC）按照100: 1的质量比溶解于碳酸缓冲液中（0.05mol/L pH9.0），4℃下避光搅拌反应12~14h，反应完成后，冷冻干燥，将冻干粉重新溶解，利用凝胶层析（Sephadex G25填料）去除未反应的FITC，分离完后，测量FITC标记的胰蛋白酶溶液的荧光强度和胰蛋白酶浓度的关系。(1) Preparation of fluorescently labeled enzymes: Dissolve trypsin and fluorescein thiocyanate (FITC) in carbonic acid buffer (0.05mol/L pH9.0) at a mass ratio of 100:1, 4 Stir the reaction at ℃ for 12~14 hours in the dark. After the reaction is completed, lyophilize and redissolve the lyophilized powder. Use gel chromatography (Sephadex G25 filler) to remove unreacted FITC. After separation, measure FITC-labeled trypsin The relationship between the fluorescence intensity of the solution and the concentration of trypsin.
（2）皮革试样裁取：取山羊皮脊背部，常规法脱毛，用圆形裁刀裁成直径50mm的皮革试样。(2) Cutting of leather samples: Take the back of the goatskin, depilate it by conventional methods, and cut it into a leather sample with a diameter of 50mm with a round knife.
（3）传质实验：调节底座调节螺栓，使气泡位于水平泡的中心，取外径40mm的上储液池和下储液池各一个，下储液池装入80%容量的实验缓冲液（0.05 mol/L pH7.4的磷酸缓冲液），粒面向上将皮革试样遮盖于下储液池上，将上储液池口对口放在皮革试样上，保持与下储液池同心，调节可调节F夹的下夹位置固定上、下储液池和皮革试样，逆时针旋转下夹调节螺丝将上下储液池及皮革试样夹紧。先将下储液池通过医用注射器注满实验缓冲液，再将上储液池1通过医用注射器注满含有FITC标记胰蛋白酶的溶液，传质30 min后，顺时针旋转下夹调节螺丝，取下皮革试样。(3) Mass transfer experiment: adjust the base adjusting bolt so that the air bubble is located in the center of the horizontal bubble, take an upper liquid storage tank and a lower liquid storage tank with an outer diameter of 40mm, and fill the lower liquid storage tank with 80% of the experimental buffer (0.05 mol/L pH7.4 phosphate buffer solution), with the grain side up, cover the leather sample on the lower reservoir, place the upper reservoir mouth-to-mouth on the leather sample, keep it concentric with the lower reservoir, and adjust Adjust the position of the lower clamp of the F clamp to fix the upper and lower reservoirs and the leather sample, and rotate the lower clamp adjustment screw counterclockwise to clamp the upper and lower reservoirs and the leather sample. First fill the lower reservoir with the experimental buffer through a medical syringe, then fill the
（4）数据采集：在皮革试样的中心取样，荧光显微镜观察皮革试样纵切片荧光强度分布情况。图3为FITC标记胰蛋白酶（FITC-Trypsin）传质30 min后皮革试样纵切片的荧光显微照片。(4) Data collection: samples were taken from the center of the leather sample, and the fluorescence intensity distribution of the longitudinal section of the leather sample was observed with a fluorescence microscope. Fig. 3 is a fluorescence micrograph of a longitudinal section of a leather sample after mass transfer with FITC-Trypsin for 30 min.
实施例3：Example 3:
（1）荧光标记酶的制备：取胰蛋白酶（Trypsin）、硫氰酸荧光素（Fluoresceinisothiocyanate，FITC）按照100: 1的质量比溶解于碳酸缓冲液中（0.05mol/L pH9.0），4℃下避光搅拌反应12-14h，反应完成后，冷冻干燥，将冻干粉重新溶解后，利用凝胶层析（Sephadex G25填料）去除未反应的FITC，分离完后，测量FITC标记的胰蛋白酶的溶液的荧光强度和浓度的关系。(1) Preparation of fluorescently labeled enzymes: Dissolve trypsin and fluorescein thiocyanate (FITC) in carbonic acid buffer (0.05mol/L pH9.0) at a mass ratio of 100:1, 4 Stir and react in the dark at ℃ for 12-14 hours. After the reaction is completed, lyophilize and redissolve the lyophilized powder, and use gel chromatography (Sephadex G25 filler) to remove unreacted FITC. After separation, measure the FITC-labeled pancreatic The relationship between fluorescence intensity and concentration of protease solution.
（2）皮革试样裁取：取山羊皮脊背部，常规法脱毛，用圆形裁刀裁成直径50mm的皮革试样。(2) Cutting of leather samples: Take the back of the goatskin, depilate it by conventional methods, and cut it into a leather sample with a diameter of 50mm with a round knife.
（3）传质实验：实验装置如图1所示，调节底座调节螺栓，使气泡位于水平泡的中心，取外径40mm的上储液池和下储液池各一个，下储液池装入80%容量的实验缓冲液（0.05mol/L pH7.4的磷酸缓冲液），粒面向上将皮革试样遮盖于下储液池上，将上储液池口对口放在皮革试样上，保持与下储液池同心，调节可调节F夹的下夹位置固定上、下储液池和皮革试样，逆时针旋转下夹调节螺丝将上下储液池及皮革试样夹紧，先将下储液池通过医用注射器注满实验缓冲液，再将上储液池1通过医用注射器注满含有FITC标记胰蛋白酶的溶液，传质30 min后，顺时针旋转下夹调节螺丝，取下皮革试样。(3) Mass transfer experiment: The experimental device is shown in Figure 1. Adjust the adjusting bolts of the base so that the air bubble is located in the center of the horizontal bubble. Take an upper liquid storage tank and a lower liquid storage tank with an outer diameter of 40mm, and install the lower liquid storage tank. Fill in 80% of the experimental buffer (0.05mol/L pH7.4 phosphate buffer), cover the leather sample on the lower reservoir with the grain side up, and place the upper reservoir mouth-to-mouth on the leather sample, keeping Concentric with the lower liquid storage tank, adjust the position of the lower clamp of the adjustable F clamp to fix the upper and lower liquid storage tanks and the leather sample, and turn the lower clamp adjustment screw counterclockwise to clamp the upper and lower liquid storage tanks and the leather sample. Fill the reservoir with the experimental buffer through a medical syringe, and then fill the
（4）数据采集：在皮革试样的中心取样，用冷冻切片机从皮革试样的粒面至肉面顺序切片，切片厚度25μm，荧光分光光度计检测（激发长光波495，发射波长525）切片荧光强度。图4为FITC标记胰蛋白酶（FITC-Trypsin）传质30 min后皮革试样不同深度横切片的荧光强度分布图。(4) Data collection: Sampling at the center of the leather sample, using a cryostat to slice sequentially from the grain surface to the meat surface of the leather sample, with a slice thickness of 25 μm, and detecting with a fluorescence spectrophotometer (excitation long light wavelength 495, emission wavelength 525) Section fluorescence intensity. Figure 4 is the fluorescence intensity distribution diagram of cross-sections of leather samples at different depths after mass transfer of FITC-Trypsin for 30 min.
实施例4：Example 4:
（1）荧光标记酶的制备：取中性蛋白酶（Dispase）、硫氰酸荧光素（Fluoresceinisothiocyanate，FITC）按照100: 1的质量比溶解于磷酸缓冲液中（0.05mol/L pH7.4），4℃下避光搅拌反应12-14h，反应完成后，冷冻干燥，将冻干粉重新溶解后，利用凝胶层析（Sephadex G25填料）去除未反应的FITC，分离完后，测量FITC标记的中性蛋白酶的溶液的荧光强度和浓度的关系。(1) Preparation of fluorescent labeling enzyme: Dissolve neutral protease (Dispase) and fluorescein thiocyanate (Fluoresceinisothiocyanate, FITC) in phosphate buffer (0.05mol/L pH7.4) according to the mass ratio of 100:1, Stir and react in the dark at 4°C for 12-14 hours. After the reaction is complete, lyophilize and redissolve the lyophilized powder. Use gel chromatography (Sephadex G25 filler) to remove unreacted FITC. After separation, measure the FITC-labeled The relationship between fluorescence intensity and concentration of dispase solution.
（2）皮革试样裁取：取山羊皮脊背部，常规法脱毛，用圆形裁刀裁成直径35mm的皮革试样。(2) Cutting of leather samples: take the back of the goatskin, depilate it by conventional methods, and cut it into a leather sample with a diameter of 35mm with a round knife.
（3）传质实验：实验装置如图1所示，调节底座调节螺栓，使气泡位于水平泡5的中心。取外径25mm的上储液池和下储液池各一个，下储液池装入80%容量的实验缓冲液（0.05mol/L pH7.4的磷酸缓冲液），粒面向上将皮革试样遮盖于下储液池上，将上储液池口对口放在皮革试样上，保持与下储液池同心，调节可调节F夹的下夹位置固定上、下储液池和皮革试样，逆时针旋转下夹调节螺丝将上下储液池及皮革试样夹紧，先将下储液池通过医用注射器注满实验缓冲液，再将上储液池通过医用注射器注满含有FITC标记中性蛋白酶的溶液，传质30min后，顺时针旋转下夹调节螺丝，取下皮革试样。(3) Mass transfer experiment: The experimental device is shown in Figure 1. Adjust the base adjusting bolt so that the air bubble is located in the center of the
（4）数据采集：在皮革试样的中心取样，荧光显微镜观察皮革试样纵切片荧光强度分布情况。图5为FITC标记中性蛋白酶（FITC-Dispase）传质30 min后皮革试样纵切片的荧光显微照片。(4) Data collection: samples were taken from the center of the leather sample, and the fluorescence intensity distribution of the longitudinal section of the leather sample was observed with a fluorescence microscope. Figure 5 is a fluorescence micrograph of a longitudinal section of a leather sample after 30 min of mass transfer with FITC-dispase.
实施例5：Example 5:
（1）荧光标记酶的制备：取中性蛋白酶（Dispase）、硫氰酸荧光素（Fluoresceinisothiocyanate，FITC）按照100: 1的质量比溶解于磷酸缓冲液中（0.05mol/L pH7.4），4℃下避光搅拌反应12-14h，反应完成后，冷冻干燥，将冻干粉重新溶解后，利用凝胶层析（Sephadex G25填料）去除未反应的FITC。分离完后，测量FITC标记的中性蛋白酶的溶液的荧光强度和浓度的关系。(1) Preparation of fluorescent labeling enzyme: Dissolve neutral protease (Dispase) and fluorescein thiocyanate (Fluoresceinisothiocyanate, FITC) in phosphate buffer (0.05mol/L pH7.4) according to the mass ratio of 100:1, Stir and react at 4°C in the dark for 12-14 hours. After the reaction is complete, freeze-dry. After re-dissolving the freeze-dried powder, use gel chromatography (Sephadex G25 filler) to remove unreacted FITC. After the separation, the relationship between the fluorescence intensity and the concentration of the FITC-labeled dispase solution was measured.
（2）皮革试样裁取：取山羊皮脊背部，常规法脱毛，用圆形裁刀裁成直径50mm的皮革试样。(2) Cutting of leather samples: Take the back of the goatskin, depilate it by conventional methods, and cut it into a leather sample with a diameter of 50mm with a round knife.
（3）传质实验：实验装置如图1所示，调节底座调节螺栓，使气泡位于水平泡的中心，取外径40mm的上储液池和下储液池各一个，下储液池装入80%容量的实验缓冲液（0.05mol/L pH7.4的磷酸缓冲液），粒面向上将皮革试样遮盖于下储液池上，将上储液池口对口放在皮革试样上，保持与下储液池同心，调节可调节F夹的下夹位置固定上、下储液池和皮革试样，逆时针旋转下夹调节螺丝将上下储液池及皮革试样夹紧，先将下储液池通过医用注射器注满实验缓冲液，再将上储液池通过医用注射器注满含有FITC标记中性蛋白酶的溶液，传质30min后，顺时针旋转下夹调节螺丝，取下皮革试样。(3) Mass transfer experiment: The experimental device is shown in Figure 1. Adjust the adjusting bolts of the base so that the air bubble is located in the center of the horizontal bubble. Take an upper liquid storage tank and a lower liquid storage tank with an outer diameter of 40mm, and install the lower liquid storage tank. Fill in 80% of the experimental buffer (0.05mol/L pH7.4 phosphate buffer), cover the leather sample on the lower reservoir with the grain side up, and place the upper reservoir mouth-to-mouth on the leather sample, keeping Concentric with the lower liquid storage tank, adjust the position of the lower clamp of the adjustable F clamp to fix the upper and lower liquid storage tanks and the leather sample, and turn the lower clamp adjustment screw counterclockwise to clamp the upper and lower liquid storage tanks and the leather sample. The liquid storage tank is filled with the experimental buffer solution through a medical syringe, and then the upper liquid storage tank is filled with a solution containing FITC-labeled neutral protease through a medical syringe. After mass transfer for 30 minutes, turn the lower clamp adjustment screw clockwise to remove the leather sample. .
（4）数据采集：在皮革试样的中心取样，用冷冻切片机从皮革试样粒面至肉面顺序切片，切片厚度25μm，荧光分光光度计检测（激发长光波495，发射波长525）切片荧光强度。图6为FITC标记中性蛋白酶（FITC-Dispase）传质30 min后皮革试样不同深度横切片的荧光强度分布图。(4) Data collection: Sampling at the center of the leather sample, using a cryostat to slice sequentially from the grain surface to the meat surface of the leather sample, with a thickness of 25 μm, and detecting the slices with a fluorescence spectrophotometer (excitation long light wavelength 495, emission wavelength 525) The fluorescence intensity. Fig. 6 is the fluorescence intensity distribution diagram of cross-sections of leather samples at different depths after mass transfer of FITC-Dispase for 30 min.
实施例6：Embodiment 6:
（1）荧光标记酶的制备：取胰蛋白酶（Trypsin）、异硫氰酸罗丹明B（Rhodamine BisothioCyanate, RBITC）按照100: 1的质量比溶解于碳酸缓冲液中（0.05mol/L pH9.0），4℃下避光搅拌反应12-14h。反应完成后，冷冻干燥。将冻干粉重新溶解后，利用凝胶层析（Sephadex G25填料）去除未反应的RBITC。分离完后，测量RBITC标记的胰蛋白酶的溶液的荧光强度和浓度的关系。(1) Preparation of fluorescently labeled enzymes: Dissolve trypsin and Rhodamine BisothioCyanate (RBITC) in carbonic acid buffer (0.05mol/L pH9.0 at a mass ratio of 100:1) ), stirred at 4°C for 12-14 hours in the dark. After the reaction was complete, it was freeze-dried. After the lyophilized powder was redissolved, unreacted RBITC was removed by gel chromatography (Sephadex G25 filler). After the separation, the relationship between the fluorescence intensity and the concentration of the RBITC-labeled trypsin solution was measured.
（2）皮革试样裁取：取脊背部山羊皮，常规脱毛，用圆形裁刀裁成直径35mm的皮革试样。(2) Cutting of leather samples: Take the back goatskin, depilate it conventionally, and cut it into a leather sample with a diameter of 35 mm with a circular cutter.
（3）传质实验：实验装置如图1所示，调节底座调节螺栓，使气泡位于水平泡的中心，取外径25mm的上储液池和下储液池各一个，下储液池装入80%容量的实验缓冲液（0.05mol/L pH7.4的磷酸缓冲液），粒面向上将皮革试样遮盖于下储液池上，将上储液池口对口放在皮革试样上，保持与下储液池同心，调节可调节F夹的下夹位置固定上、下储液池和皮革试样，逆时针旋转下夹调节螺丝将上下储液池及皮革试样夹紧；先将下储液池通过医用注射器注满实验缓冲液，再将上储液池通过医用注射器注满含有RBITC标记胰蛋白酶的溶液，传质30 min后，顺时针旋转下夹调节螺丝，取下皮革试样。(3) Mass transfer experiment: The experimental device is shown in Figure 1. Adjust the adjusting bolts of the base so that the air bubble is located in the center of the horizontal bubble. Take an upper liquid storage tank and a lower liquid storage tank with an outer diameter of 25mm, and install the lower liquid storage tank. Fill in 80% of the experimental buffer (0.05mol/L pH7.4 phosphate buffer), cover the leather sample on the lower reservoir with the grain side up, and place the upper reservoir mouth-to-mouth on the leather sample, keeping Concentric with the lower liquid storage tank, adjust the position of the lower clamp of the adjustable F clamp to fix the upper and lower liquid storage tanks and the leather sample, and turn the lower clamp adjustment screw counterclockwise to clamp the upper and lower liquid storage tanks and the leather sample; Fill the storage tank with the experimental buffer through a medical syringe, and then fill the upper storage tank with a solution containing RBITC-labeled trypsin through a medical syringe. After mass transfer for 30 minutes, turn the lower clamp adjustment screw clockwise to remove the leather sample .
（4）荧光标记酶的制备：在皮革试样的中心取样，用冷冻切片机从皮革试样的粒面至肉面顺序切片，切片厚度25μm，荧光分光光度计检测（激发长光波495，发射波长525）切片荧光强度。图7为RBITC标记胰蛋白酶（RBITC-Trypsin）传质30 min后皮革试样不同深度横切片的荧光强度分布图。(4) Preparation of fluorescently labeled enzyme: take a sample from the center of the leather sample, slice it sequentially from the grain surface to the flesh surface of the leather sample with a cryostat, and slice the thickness of 25 μm, and detect it with a fluorescence spectrophotometer (excitation long light wave 495, emission wavelength 525) slice fluorescence intensity. Fig. 7 is the fluorescence intensity distribution diagram of cross-sections of leather samples at different depths after mass transfer of RBITC-Trypsin for 30 min.
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| CN201910007178.1AActiveCN109490274B (en) | 2019-01-04 | 2019-01-04 | Experimental device for researching unidirectional mass transfer of enzyme in leather and application method |
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Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH05223806A (en)* | 1992-02-13 | 1993-09-03 | Hitachi Ltd | Glycoprotein analyzer |
| JPH0961355A (en)* | 1995-08-30 | 1997-03-07 | Mitsubishi Heavy Ind Ltd | Optical axis moving type fluorometer and measuring method |
| US5804395A (en)* | 1995-12-01 | 1998-09-08 | The United States Of America As Represented By The Secretary Of The Navy | Fluorescence polarization assays of enzymes and substrates therefore |
| CN101011237A (en)* | 2007-02-01 | 2007-08-08 | 中国药科大学 | Pharmaceutical in vivo dynamics characteristic-nondestructive in situ monitoring system and monitoring method |
| CN101074952A (en)* | 2007-06-22 | 2007-11-21 | 中国科学院植物研究所 | Immune fluorescent marking method for naked plant pollen tube microtubule skeleton and its use |
| CN101271070A (en)* | 2008-05-09 | 2008-09-24 | 东北大学 | Microfluidic capillary electrophoresis liquid core waveguide fluorescence detection device |
| CN102495037A (en)* | 2011-12-07 | 2012-06-13 | 南京农业大学 | Fluorescent tracing method of assimilation substance unloading path of phloem in pear fruit |
| CN103243149A (en)* | 2013-05-20 | 2013-08-14 | 吉林大学 | Method for rapidly determining trypsin and aprotinin combination mechanism |
Family Cites Families (29)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0697221B2 (en)* | 1987-05-18 | 1994-11-30 | 麒麟麦酒株式会社 | Method for measuring organic concentration and apparatus used for the method |
| WO1994000602A1 (en)* | 1992-06-29 | 1994-01-06 | Sensor Technologies, Inc. | Method and device for detecting and quantifying substances in body fluids |
| US7364565B2 (en)* | 2001-07-27 | 2008-04-29 | Ramot At Tel Aviv University Ltd. | Controlled enzymatic removal and retrieval of cells |
| JP3896272B2 (en)* | 2001-10-23 | 2007-03-22 | 財団法人日本紡績検査協会 | Judgment method of natural leather products |
| AU2003225165A1 (en)* | 2002-04-26 | 2003-11-10 | Board Of Trustees Of The Leland Stanford Junior University | System and method of measuring molecular interactions |
| JP2005080535A (en)* | 2003-09-05 | 2005-03-31 | Olympus Corp | Method for detecting reaction between nucleic acid base and helicase |
| WO2006114130A1 (en)* | 2005-04-26 | 2006-11-02 | Agilent Technologies, Inc. | Enzymes with modified amino acids |
| US20070190593A1 (en)* | 2005-09-20 | 2007-08-16 | Arizona Board Of Regents, Acting For And On Behalf Of Arizona State University | Methods for detecting and characterizing microorganisms |
| JP4763485B2 (en)* | 2006-03-15 | 2011-08-31 | 株式会社日立ハイテクノロジーズ | Fluorescence detection device |
| US7498551B2 (en)* | 2006-10-06 | 2009-03-03 | Los Alamos National Security, Llc | Apparatus and method for tracking a molecule or particle in three dimensions |
| CN100552434C (en)* | 2007-01-09 | 2009-10-21 | 浙江大学 | Fluorescence Spectroscopic Analysis of Immobilized Enzyme Conformation |
| US20100323945A1 (en)* | 2007-01-11 | 2010-12-23 | Novozymes A/S | Particles Comprising Active Compounds |
| CN101730746A (en)* | 2007-07-06 | 2010-06-09 | 帕普斯特许可有限两合公司 | Determination of the activity of proteases |
| CN201186115Y (en)* | 2008-04-11 | 2009-01-28 | 江苏省华建建设股份有限公司 | Drop-prevention wall-clamping device for aloft work |
| JP2011047802A (en)* | 2009-08-27 | 2011-03-10 | Olympus Corp | Concentration measuring method of target material |
| JP2011169834A (en)* | 2010-02-19 | 2011-09-01 | Kansai Bunri Sogo Gakuen | Method and system for detecting lipid vesicle derived from pathogenic gram-negative bacterium |
| DE102010030069A1 (en)* | 2010-06-15 | 2011-12-15 | Agilent Technologies Inc. | Liquid chromatography fluorescence detector for detecting component of fluid sample in e.g. life science apparatus, for separating component of fluid sample, has detection channel detecting fluorescence radiation |
| CN102323308B (en)* | 2011-06-14 | 2013-09-11 | 北京科技大学 | Device and method for researching hydrogen permeation behavior of metal subjected to dead-load pulling stress in gaseous medium |
| US9879305B2 (en)* | 2012-05-02 | 2018-01-30 | Alberto Morales Villagran | Device for online measurement of neurotransmitters using enzymatic reactors |
| JPWO2014034275A1 (en)* | 2012-08-30 | 2016-08-08 | 株式会社日立ハイテクノロジーズ | Nucleic acid analyzer |
| CN103439500B (en)* | 2013-08-27 | 2015-01-28 | 陕西科技大学 | Visible tracking and detecting method for protease in leather treatment process |
| CN103525954B (en)* | 2013-10-28 | 2015-07-22 | 四川大学 | Immobilized composite unhairing enzyme with leather-making secondary waste as carrier and preparation method thereof |
| EP3169795B1 (en)* | 2014-07-17 | 2019-11-20 | Sano LLC | Preparation of stabilizer-free gold sols and their configuration into a gelatin-coated protease-indicating homogeneous (one step) assay |
| GB201418007D0 (en)* | 2014-10-10 | 2014-11-26 | Xeros Ltd | Animal skin substrate Treatment apparatus and method |
| RU2567510C1 (en)* | 2014-11-17 | 2015-11-10 | Федеральное государственное бюджетное образовательное учреждение высшего профессионального образования "Восточно-Сибирский государственный университет технологий и управления" | Method for measuring mass transfer rate in capillary-porous bodies |
| EP3073246B1 (en)* | 2015-03-27 | 2019-08-14 | National Institute of Biology | Method and system for simultaneous detection of micro-particle concentration in suspension and their morphological and physiological traits |
| CN207326783U (en)* | 2017-07-10 | 2018-05-08 | 上海新月工具有限公司 | A kind of multi-functional F folders |
| CN207415218U (en)* | 2017-10-20 | 2018-05-29 | 浙江光辉工具有限公司 | It is a kind of can realizing self disassembling F type fixtures |
| CN209460142U (en)* | 2019-01-04 | 2019-10-01 | 齐鲁工业大学 | An experimental device for studying one-way mass transfer of enzymes in leather |
- 2019
- 2019-01-04CNCN201910007178.1Apatent/CN109490274B/enactiveActive
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH05223806A (en)* | 1992-02-13 | 1993-09-03 | Hitachi Ltd | Glycoprotein analyzer |
| JPH0961355A (en)* | 1995-08-30 | 1997-03-07 | Mitsubishi Heavy Ind Ltd | Optical axis moving type fluorometer and measuring method |
| US5804395A (en)* | 1995-12-01 | 1998-09-08 | The United States Of America As Represented By The Secretary Of The Navy | Fluorescence polarization assays of enzymes and substrates therefore |
| CN101011237A (en)* | 2007-02-01 | 2007-08-08 | 中国药科大学 | Pharmaceutical in vivo dynamics characteristic-nondestructive in situ monitoring system and monitoring method |
| CN101074952A (en)* | 2007-06-22 | 2007-11-21 | 中国科学院植物研究所 | Immune fluorescent marking method for naked plant pollen tube microtubule skeleton and its use |
| CN101271070A (en)* | 2008-05-09 | 2008-09-24 | 东北大学 | Microfluidic capillary electrophoresis liquid core waveguide fluorescence detection device |
| CN102495037A (en)* | 2011-12-07 | 2012-06-13 | 南京农业大学 | Fluorescent tracing method of assimilation substance unloading path of phloem in pear fruit |
| CN103243149A (en)* | 2013-05-20 | 2013-08-14 | 吉林大学 | Method for rapidly determining trypsin and aprotinin combination mechanism |
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|---|---|
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