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A method and application of enzymatic hydrolysis for preparing low molecular weight kappa-carrageenan potassium

A low-molecular-weight, potassium-carrageenan technology, used in pharmaceutical formulations, organic active ingredients, medical preparations containing active ingredients, etc. Potassium blood pressure research and other issues, to achieve the effect of high purity and high potassium ion content

Active Publication Date: 2021-06-11
BRILLIANCE BIO TECH CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, there is almost no research on low molecular weight κ-carrageenan potassium in reducing blood pressure, and there is no research on the preparation process of κ-carrageenan potassium in any molecular weight range

Method used

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  • A method and application of enzymatic hydrolysis for preparing low molecular weight kappa-carrageenan potassium
  • A method and application of enzymatic hydrolysis for preparing low molecular weight kappa-carrageenan potassium
  • A method and application of enzymatic hydrolysis for preparing low molecular weight kappa-carrageenan potassium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] 1. Preparation of κ-carrageenase

[0056] According to the complete gene sequence of κ-carrageenase from Zobelliasp.ZM-2, design upstream primer F1: 5'-CGGGGTACCATGACAAAACTAAAGTTTAACGGC-3', SEQ ID NO: 2; and downstream primer R1: 5'-ATAAGAATGCGGCCGCTTAAGCCGAAGTTCCGGGCG-3' , SEQ ID NO: 3;

[0057] The amplification process uses genomic DNA as a template and uses high-fidelity pfu polymerase (Takara) to amplify the target gene. The amplification reaction system (50 μL) is shown in Table 1.

[0058] Table 1 Amplification reaction system

[0059]

[0060] The PCR amplification program was as follows: pre-denaturation at 98°C, 2min; denaturation at 94°C, 30s; annealing at 56°C, 30s; extension at 72°C, 1min; final extension at 72°C, 10min; the number of cycles was 30 times.

[0061] After PCR amplification, the PCR product purification kit was used to purify, and after electrophoresis imaging, the target band and the empty vector pPIC9K were subjected to AvrII and NotI ...

Embodiment 2

[0072] 1. Preparation of κ-carrageenase

[0073] In the same way, the recombinant strain was inoculated into YPD medium, cultured at 30°C for 24h; inoculated with 1% inoculum in BMGY fermentation broth culture medium for 40h, added 1.2% methanol by volume, cultivated at 22°C for 24h, added 1.2% methanol , cultivated at 22°C for 24h, added 1.2% methanol, cultivated at 22°C for 24h, added 1.2% methanol, cultivated at 22°C for 24h, centrifuged to obtain the supernatant which was the crude enzyme solution of κ-carrageenase.

[0074] 2. Enzyme hydrolysis

[0075] Prepare 1% κ-carrageenan aqueous solution (same as in Example 1), stir and dissolve in a water bath at 40°C; centrifuge the crude enzyme solution at 10,000 r / min for 10 minutes to get the supernatant, add κ-carrageenan in a volume ratio of 2% In the solution, continue to maintain the enzymatic hydrolysis at 40°C in a water bath for 7 hours.

[0076] 3. Alcohol precipitation

[0077] Centrifuge the enzymatic solution, ta...

Embodiment 3

[0092] Example 3 Study on the Antihypertensive Effect of Low Molecular Weight κ-Carrageenan Potassium on Spontaneously Hypertensive Rats

[0093] SPF grade male spontaneously hypertensive rats (SHR) at the age of 9 weeks were selected, randomly divided into groups according to blood pressure and body weight, 10 in the blank group, orally administered distilled water; 6 in the experimental group, orally administered with the low molecular weight κ-carrageenan of Example 1 Potassium, gavage dose is 600mg / kgbw / day; captopril group 6, captopril gavage dose is 10mg / kgbw / day; measure systolic blood pressure (SBP) and heart rate once a week, weigh 2 times (for calculating the gavage volume).

[0094]The effect of low molecular weight κ-carrageenan potassium on the SBP of SHR is shown in Table 2. It can be seen that after the third week of gavage, the systolic blood pressure of the experimental group has been significantly reduced. There is a significant difference (P=0.034), indicat...

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Abstract

The present invention disclosed a method of enzyme disintegration to prepare low molecular weight κ‑ Kara gum, including the following steps: (1) In a water bath pot at 40‑50 ° C, the mix of κ‑ 本 gum powder is matched with a mass concentration of 0.5‑1.5 %The scallion gel solution, stir well, add κ‑cara gel enzyme, and mix the enzyme disintegration 6‑8h; (2) the enzymatic solution for meter sinking to obtain the low molecular meal scalcal oligosaccharide;制 Kara gum oligosaccharides are prepared into a solution with a mass concentration of 4 to10 %, mixed with a potassium hydraulic solution with a quality concentration of 0.1 % ‑2 %, and washed the alcohol to the mixture to obtain a low -molecular amount of κ‑ κ‑ κ‑ κ‑ potassiumEssenceThe method of the present invention is simple, the conditions are mild, the molecular weight of the low molecular weight of the low molecular measuring caraxia glue is stable in a smaller range, and the combination of potassium ions is high. It can be added as a food additive as a health product.Polycpense is used as a drug to prevent or treat hypertension.

Description

technical field [0001] The invention relates to the technical field of preparation of low-molecular-weight κ-carrageenan potassium, and more specifically relates to a method and application for enzymatically preparing low-molecular-weight κ-carrageenan potassium. Background technique [0002] As the original producer of organic matter in the ocean, marine algae have special carbohydrates and minerals that are indispensable for normal human physiological metabolism. Seaweed polysaccharide refers to various high-molecular carbohydrates in seaweed with high viscosity and coagulation ability. With the vigorous development of marine drugs in recent years, the research on seaweed polysaccharides has been paid more and more attention. κ-carrageenan is an extract of red algae, one of the three major marine algae. With its unique physical and chemical properties and wide range of uses, it has attracted the attention of contemporary biology, biochemistry and medicine, and has become ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P19/14C12P19/04A61K31/737A61P9/12
CPCC12P19/24A61P9/12A61K31/737C12P19/14C12P19/04
Inventor 牟海津郁东兴陈梦郁万帅韩振莲
Owner BRILLIANCE BIO TECH CO LTD
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