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Method for extracting bromelain ananase through chitosan carboxylation derivative

A technology of bromelain and chitosan, applied in the direction of hydrolytic enzymes, etc., can solve the problems of less labor, poor reusability, and easy contamination of ultrafiltration membranes

Inactive Publication Date: 2012-07-18
AGRI PRODS PROCESSING RES INST CHINESE ACAD OF TROPICAL AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

It includes the following steps: raw material-cleaning-coarse crushing-fine crushing-pretreatment-juicing-centrifugation-freezing-filtration-ultrafiltration-drying-finished product. This process relies on advanced separation and extraction methods, which requires less labor and less enzyme activity. The disadvantage is that the pineapple juice is directly ultra-filtered without sugar and miscellaneous cells, and the ultra-filtration membrane is easily polluted. It must be cleaned before and after each use. Frequent replacement of membrane blocks, high production costs

Method used

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  • Method for extracting bromelain ananase through chitosan carboxylation derivative

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1 (ultrafiltration collects bromelain)

[0039] (1) Chitosan swelling that quality is 100g is in isobutanol, and the suspension that makes mass concentration is 0.8%, adds the saturated NaOH solution that contains 65g solute wherein to it while stirring, after adding, continue at room temperature Stir for 30 minutes to obtain a chitosan raw material solution; place the solution in a 65°C water bath, add 110 g of solid chloroacetic acid to it while slowly stirring, and react after 2.5 hours, filter, collect the precipitate, and wash repeatedly with ethanol. Dry at 60°C to obtain chitosan carboxylated derivatives.

[0040] (2) Weigh 20kg of pineapple peel, wash it, squeeze it, and filter to remove large particles of impurities to obtain 11L of pineapple juice, which is placed in a refrigerated tank at 8°C, and powdered NaHSO is added thereto 3 0.0033g, cysteine ​​0.0006g, EDTA (ethylenediaminetetraacetic acid) 0.0049g, stir slowly and evenly.

[0041] (3) get...

Embodiment 2

[0044] Embodiment 2 (ultrafiltration collects bromelain)

[0045](1) Chitosan swelling that quality is 100g is in isobutanol, and the suspension that makes mass concentration is 1.1%, adds the saturated NaOH solution that contains 80g solute wherein to it while stirring, after adding, continue at room temperature Stir for 30 minutes to obtain chitosan raw material solution. Place the solution in a water bath at 75°C, add 130g of solid chloroacetic acid to it while stirring slowly, react for 3.0 hours, filter, collect the precipitate, wash repeatedly with ethanol, and dry at 70°C to obtain chitosan carboxylated derivatives.

[0046] (2) Weigh 30kg of pineapple peel, wash it, squeeze it, and filter to remove large particles of impurities to obtain 16L of pineapple juice, which is placed in a refrigerated tank at 14°C, and powdered NaHSO is added thereto 3 0.0064g, cysteine ​​0.0010g, EDTA 0.0088g, stir slowly evenly.

[0047] (3) get the chitosan carboxylated derivative 56g o...

Embodiment 3

[0050] Embodiment 3 (ultrafiltration collects bromelain)

[0051] (1) the chitosan swelling that quality is 100g is in isobutanol, and the suspension that makes mass concentration is 0.9%, adds the saturated NaOH solution that contains 70g solute wherein to it while stirring, after adding, continue at room temperature Stir for 30 minutes to obtain chitosan raw material solution. Place the solution in a water bath at 70°C, add 120 g of solid chloroacetic acid into it with slow stirring, react for 2.8 hours, filter, collect the precipitate, wash repeatedly with ethanol, and dry at 65°C to obtain carboxylated chitosan derivative.

[0052] (2) Weigh 50kg of pineapple peel, wash it, squeeze it, and filter to remove large particles of impurities to obtain 28L of pineapple juice, which is placed in a refrigerated tank at 10°C, and powdered NaHSO is added thereto 3 0.0098g, cysteine ​​0.0017g, EDTA 0.014g, stir slowly evenly.

[0053] (3) Get 76g of chitosan carboxylated derivative...

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Abstract

The invention provides a method for extracting bromelain ananase through a chitosan carboxylation derivative, which comprises the following steps of: (1) preparing the chitosan carboxylation derivative; (2) carrying out preprocessing on pineapple juice; (3) adsorbing the bromelain ananase; (4) eluting the bromelain ananase; and (5) refining and drying the bromelain ananase. The method for extracting bromelain ananase through the chitosan carboxylation derivative has the advantages that: (1) the obtained enzyme has high activity, and is safe and reliable; (2) the process has less pollution to an ultrafiltration membrane, and the production efficiency is high; (3) the production of the bromelain ananase is not limited by pineapple harvest time; (4) two products of the bromelain ananase and the pineapple juice can be obtained simultaneously; and (5) the process is flexible, the bromelain ananase can be collected through ultrafiltration or salting out, and when the salting out method is used, the volume of the bromelain ananase solution is only 1 / 20 of the original pineapple juice, so that the used salt is greatly reduced.

Description

【Technical field】 [0001] The invention relates to a method for extracting bromelain, in particular to a method for extracting bromelain with chitosan carboxylated derivatives. 【Background technique】 [0002] Bromelain (bromelain for short) is a general term for a class of polypeptide hydrolase extracted from pineapple plants, mainly present in pineapple stems and fruits. According to different extraction parts, it is divided into stem bromelain (EC 3.4.22.32) and fruit bromelain (EC 3.4.22.33). Fruit bromelain is extracted from pineapple peel and fruit, and stem bromelain is extracted from pineapple stems. The present invention is aimed at the separation and extraction of fruit bromelain. [0003] Bromelain is a sulfhydryl protease with strong hydrolytic activity and is widely used in food and biochemical fields. At the same time, because it can strengthen the hydrolysis of fibrin in the body, dissolve the fibrin and blood clots blocked in the tissue, promote the penetrat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/50
Inventor 韩志萍黄茂芳李普旺
Owner AGRI PRODS PROCESSING RES INST CHINESE ACAD OF TROPICAL AGRI SCI
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