Earthworm composite multienzyme and preparation method thereof

An earthworm and lipase technology, applied in the fields of biochemical equipment and methods, enzymes, enzymes, etc., can solve problems such as the lack of complex multi-enzymes

Inactive Publication Date: 2009-11-04
珠海博康药业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0002] Existing published patent literature technolog...

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] (1) Wash fresh and live earthworms, homogenize, stir, and centrifuge in a buffer solution at 5-15 degrees Celsius and pH=4-8, and discard the sediment to obtain a crude extract.

[0016] (2) The above-mentioned crude extract is heated to 40±2 degrees, kept warm for 2-4 hours, cooled to 5-15 degrees, and acetone is added to the filtrate after filtration.

[0017] (3) The above-mentioned precipitate was washed and dissolved with buffer solution, and the supernatant was obtained by discarding the precipitate.

[0018] (4) Add mg, Cu, Zn and protective agent to the supernatant under stirring, then centrifuge and discard the precipitate.

[0019] (5) Add ammonium sulfate solution to the supernatant, and centrifuge to collect the precipitate; dissolve the above precipitate in the buffer, and then dialyze until there is no SO 2- 4 .

[0020] (6) The dialysate is collected on a molecular sieve chromatography column to collect the eluate.

[0021] (7) Ultrafiltration and con...

Embodiment 2

[0024] (1) Wash the air-dried earthworms, soak them at 10-15 degrees Celsius for 4-6 hours, homogenize them in a buffer solution with a pH of 4-8, stir them, centrifuge them, and discard the sediment to obtain a crude extract.

[0025] (2) The above-mentioned crude extract is heated to 40±2 degrees, kept warm for 2-4 hours, cooled to 5-15 degrees, and acetone is added to the filtrate after filtration.

[0026] (3) The above-mentioned precipitate was washed and dissolved with buffer solution, and the supernatant was obtained by discarding the precipitate.

[0027] (4) Add mg, Cu, Zn and protective agent to the supernatant under stirring, then centrifuge and discard the precipitate.

[0028] (5) Add ammonium sulfate solution to the supernatant, and centrifuge to collect the precipitate; dissolve the above precipitate in the buffer, and then dialyze until there is no SO 2- 4 .

[0029] (6) The dialysate is collected on a molecular sieve chromatography column to collect the elu...

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PUM

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Abstract

The invention relates to a composite multienzyme prepared from earthworms and a preparation method thereof. The composite multienzyme comprises various proteolytic enzymes, nuclease, lipoidase, muramidase, phytase, amylase, superoxide dismutase, catalase, cellulose, and the like. The preparation method comprises the following steps: using the earthworms as materials; protecting homogenate at low temperature by a buffer solution; centrifugally separating; washing; precipitating and separating by adding ammonium sulphate; dialyzing to remove SO4; carrying out chromatography by a molecular sieve; and ultrafiltering, concentrating and drying in vacuum at low temperature. The composite multienzyme prepared by purification has the characteristics of high efficiency and multifunction in the aspects of biomacromolecule decomposition and substrate utilization. The preparation method has low cost, and the composite multienzyme has high activity. The functions of all enzymes or partial enzymes of the composite multienzyme can be utilized to achieve a corresponding use aim, thus the composite multienzyme has wide application range.

Description

technical field [0001] The invention provides a compound multienzyme prepared from earthworms and a preparation method thereof. The compound multi-enzyme comprises: various proteases, nucleases, lipases, lysozymes, phytases, amylases, superoxide dismutases, catalases, cellulases and the like. Its preparation includes method steps such as earthworm treatment, homogenization, centrifugal separation, extraction, dialysis, molecular sieve chromatography and the like. The earthworm compound multi-enzyme can be used in technical fields such as food, feed, cosmetics, medicine, biological products, biological culture, animal breeding, and biological engineering. Background technique [0002] In the existing disclosed patent literature technology, there is no compound multienzyme prepared from earthworms. Enzyme preparations have a wide range of uses in the biological field, and their sources, compositions, targets, and preparation methods have their own characteristics. Earthworm...

Claims

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Application Information

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IPC IPC(8): C12N9/00
Inventor 孙启达陈东古练权钟世勇房玮琳
Owner 珠海博康药业有限公司
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