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Tyrosinase inhibitor and preparation method thereof

A technology of tyrosinase and inhibitor, which is applied in the field of inhibitor and its preparation, can solve the problems of corrosion, and achieve the effects of less time consuming, high purification multiple, high enzyme activity, and simple operation

Inactive Publication Date: 2012-02-01
DONGHUA UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

These compounds can quickly disintegrate melanin tissue, but because of their corrosiveness, cytotoxicity and allergies, cytotoxicity and allergies, etc., they are extremely dangerous and have been banned in many countries' health regulations

Method used

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  • Tyrosinase inhibitor and preparation method thereof
  • Tyrosinase inhibitor and preparation method thereof
  • Tyrosinase inhibitor and preparation method thereof

Examples

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Embodiment 1

[0027] Using tyrosinase molecules as targeting molecules, a large number of phages with different affinities were screened, and the specific steps were as follows:

[0028] (1) Place 100 μg / ml tyrosinase solution in a microwell plate and incubate overnight at 4°C, pour off the coating solution, and block with blocking solution, then wash the plate with TBS buffer for 6 times, then add Shake 10 μl phage at room temperature for 40 minutes, wash the plate 6 times with TBS buffer, add 100 μl elution buffer for competitive elution for 35 minutes;

[0029] (2) Transfer the eluted phage to a centrifuge tube, store at 4°C, and take 1 μl for titer test. The remainder was used for phage amplification.

[0030] Amplify the screened affinity phage, the specific steps are as follows:

[0031] (1) Add the eluate from the previous round to 50ml of ER2738 culture, shake slowly at 70rpm at 37°C for 1h; (2) Take the bacterial solution, centrifuge at 5000rpm for 10min, and resuspend the pellet...

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Abstract

The invention relates to a tyrosinase inhibitor and a preparation method thereof. The sequence of the tyrosinase inhibitor is Ile-Gln-Ser-Pro-His-Phe-Phe. The preparation method comprises the following steps: (1) with tyrosinase molecules as targeting molecules, screening considerable bacteriophages having different affinity for the tyrosinase molecules; (2) amplifying screened bacteriophages with affinity for subsequent biospanning; (3) determining affinity of a plurality of screened monoclonal bacteriophages and selecting monoclonal bacteriophages with strongest bonding force for DNA sequencing and synthesis of specific ligands. The tyrosinase inhibitor provided in the invention has a good inhibitory effect; the tyrosinase inhibitor is an endogenous substance of human bodies and has good biocompatibility and low cytotoxicity, and therefore, the tyrosinase inhibitor is more suitable for being applied in fields of whitening cosmetics, preservation of fruit and vegetable, biopesticides, etc. The preparation method has the advantages of simple operation and little time consumption; the prepared tyrosinase inhibitor has a high purification fold and high enzyme activity.

Description

technical field [0001] The invention belongs to the field of inhibitors and preparation thereof, in particular to a tyrosinase inhibitor and a preparation method thereof. Background technique [0002] Tyrosinase (EC1.14.18.1) is a class of copper-containing metalloenzymes, which widely exist in bacteria, fungi, gymnosperms, angiosperms, mammals and other organisms, and exist in various levels of phylogenetic stages in the biological kingdom. In the human body, it is a key enzyme involved in the synthesis of melanin. Inhibiting its activity can inhibit the production of melanin. Therefore, it has become a research and development trend in the pharmaceutical and cosmetic industries to find effective polypeptide tyrosinase inhibitors that have no adverse reactions to the human body. The browning of fruits and vegetables during harvesting, storage and processing is mainly the result of the action of tyrosinase. It is a desirable method to develop fruit and vegetable preservati...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06C07K1/14C12Q1/70C12Q1/30C12Q1/26
Inventor 朱利民汪雯聂华丽陈天翔
Owner DONGHUA UNIV
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