阅读说明：本技术 一种高盐发酵红曲黄色素制备方法 (Preparation method of high-salt fermented monascus yellow pigment ) 是由 陈功 赵露 赵喜红 胡婷 于 2019-10-16 设计创作，主要内容包括：本发明公开了一种高盐发酵红曲黄色素制备方法,包括如下步骤：步骤1：制备含高盐的基本培养基,且基本培养基中的盐含量不低于20-50g/L；步骤2：取红曲菌种子液接种于步骤1所制备基本培养基中进行好氧发酵4-7d；步骤3：从步骤2所得的发酵液进行固液分离,分别从清液中得到胞外红曲黄色素,从固态物中通过乙醇提取分离法获得胞内红曲黄色素,其中,胞外红曲黄色素与胞内红曲黄色素总和即为红曲黄色素产品。本发明通过采用高盐的极端环境中进行发酵,其能促进红曲菌代谢生成红曲黄色素,如此可显著的提高红曲黄色素的产量,如此可考虑直接采用海水或浓缩的海水来配制培养基进行发酵生产,使得整个生产方法生产成本低,且环境友好,同时产量高。(The invention discloses a preparation method of high-salt fermented monascus yellow pigment, which comprises the following steps of 1, preparing a basic culture medium containing high salt, wherein the salt content in the basic culture medium is not lower than 20-50g/L, 2, inoculating monascus seed liquid into the basic culture medium prepared in the step 1 for aerobic fermentation for 4-7d, and 3, performing solid-liquid separation on fermentation liquid obtained in the step 2, respectively obtaining extracellular monascus yellow pigment from clear liquid, and obtaining intracellular monascus yellow pigment from solid substances through an ethanol extraction separation method, wherein the sum of the extracellular monascus yellow pigment and the intracellular monascus yellow pigment is a monascus yellow pigment product.)

1, A preparation method of high-salt fermented monascus yellow pigment, which is characterized by comprising the following steps:

step 1: preparing a basic culture medium containing high salt, wherein the salt content in the basic culture medium is not less than 20-50 g/L;

step 2: inoculating monascus seed liquid into the basic culture medium prepared in the step 1 for aerobic fermentation for 4-7 days;

and step 3: and (3) performing solid-liquid separation on the fermentation liquor obtained in the step (2), respectively obtaining extracellular monascus yellow pigment from clear liquid, and obtaining intracellular monascus yellow pigment from the solid through an ethanol extraction separation method, wherein the sum of the extracellular monascus yellow pigment and the intracellular monascus yellow pigment is a monascus yellow pigment product.

2. The method for preparing monascus yellow pigment through high-salt fermentation according to claim 1, wherein the salt in the step 1 is NaCl.

3. The method for preparing monascus yellow pigment through high-salt fermentation according to claim 2, wherein the components left in the minimal medium in the step 1 are respectively as follows: glucose, (NH)₄)₂SO₄、KH₂PO₄、KCl、MgSO₄·7H₂O、FeSO₄·7H₂O、ZnSO₄·7H₂O、MnSO₄·H₂O and water, and the concentration of glucose in the minimal medium is 50g/L, (NH)₄)₂SO₄The concentration of (A) is 5g/L, KH₂PO₄Has a concentration of 5g/L, KCl concentration of 0.5g/L, MgSO₄·7H₂The concentration of O is 0.5g/L, FeSO₄·7H₂O concentration of 0.01g/L, ZnSO₄·7H₂O concentration of 0.01g/L, MnSO₄·H₂The concentration of O was 0.03 g/L.

4. The method for preparing Monascus yellow pigment according to any one of claims 1-3 and , wherein the Monascus is Monascus ruber deposited in the china committee for culture collection management of microorganisms with the collection number of CGMCC 10910.

5. The method for preparing monascus yellow pigment through high-salt fermentation according to claim 4, wherein the inoculation amount of the monascus seed liquid in the step 2 is 5-10% of the volume of the basic culture medium.

6. The method for preparing monascus yellow pigment through high-salt fermentation according to claim 5, wherein the monascus seed solution is prepared by the following steps: taking the strain to coat the strain in a plate culture medium for culturing for 4-7d, taking bacterial colonies on the plate after culturing, adding the bacterial colonies into a seed culture medium for culturing for 24-36h, and the adding amount of the bacterial colonies in the seed culture medium is 5-10 per 50 mL.

7. The method of claim 6, wherein the culture medium comprises glucose, potato, agar and water, wherein the glucose concentration is 20g/L, the potato concentration is 200g/L, and the agar concentration is 20 g/L.

8. The method for preparing monascus yellow pigment through high-salt fermentation according to claim 6, wherein the components in the seed culture medium are glucose, yeast extract, peptone, KCl and FeSO₄·7H₂O、KH₂PO₄And water, wherein the concentration of glucose is 20g/L, the concentration of yeast extract is 3g/L, the concentration of peptone is 10g/L, the concentration of KCl is 0.5g/L, FeSO₄·7H₂The concentration of O is 0.01g/L, KH₂PO₄The concentration of (2) is 4 g/L.

9. The method for preparing monascus yellow pigment through high-salt fermentation according to claim 4, wherein the aerobic fermentation in the step 2 is performed in a constant-temperature shaking table, the fermentation temperature is 25-35 ℃, and the shaking frequency is 150-250 rpm.

10. The method for preparing monascus yellow pigment through high-salt fermentation according to claim 9, wherein the fermentation temperature in the aerobic fermentation in the step 2 is 30 ℃, and the oscillation frequency is 180 rpm.

Technical Field

The invention belongs to the field of fermentation, and particularly relates to a preparation method of high-salt fermented monascus yellow pigment.

Background

Monascus is filamentous fungi, which has been studied for thousands of years in Asian countries, monascus pigment is a -class natural mixed pigment with a polyketone structure and mainly comprises three major classes of red, orange and yellow, and recent research reports indicate that monascus pigment, especially monascus yellow pigment, has antibacterial, anticancer, antioxidant, lipid-lowering and other functional activities, and can be widely used in the fields of food and medicine .

The monascus pigment is obtained by monascus liquid fermentation, which is the key point of research, and most reports focus on optimizing the composition of a culture medium and fermentation conditions, so as to improve the content of the monascus pigment, but the improvement amount is limited.

Disclosure of Invention

In order to solve the technical problems, the invention aims to provide preparation methods with high yield and simple separation for high-salt fermented monascus yellow pigment.

In order to realize the purpose, the technical scheme of the invention is that the preparation method of high-salt fermentation monascus yellow pigment comprises the following steps:

step 1: preparing a basic culture medium containing high salt, wherein the salt content in the basic culture medium is not less than 20-50 g/L;

step 2: inoculating monascus seed liquid into the basic culture medium prepared in the step 1 for aerobic fermentation for 4-7 days;

and step 3: and (3) performing solid-liquid separation on the fermentation liquor obtained in the step (2), respectively obtaining extracellular monascus yellow pigment from clear liquid, and obtaining intracellular monascus yellow pigment from the solid through an ethanol extraction separation method, wherein the sum of the extracellular monascus yellow pigment and the intracellular monascus yellow pigment is a monascus yellow pigment product.

Wherein, the salt in the step 1 is NaCl.

Wherein, the components remained in the minimal medium in the step 1Respectively, the following steps: glucose, (NH)₄)₂SO₄、KH₂PO₄、KCl、MgSO₄·7H₂O、FeSO₄·7H₂O、ZnSO₄·7H₂O、MnSO₄·H₂O and water, and the concentration of glucose in the minimal medium is 50g/L, (NH)₄)₂SO₄The concentration of (A) is 5g/L, KH₂PO₄Has a concentration of 5g/L, KCl concentration of 0.5g/L, MgSO₄·7H₂The concentration of O is 0.5g/L, FeSO₄·7H₂O concentration of 0.01g/L, ZnSO₄·7H₂O concentration of 0.01g/L, MnSO₄·H₂The concentration of O was 0.03 g/L.

Wherein the Monascus ruber is preserved in the China general microbiological culture Collection center of China Committee for culture Collection of microorganisms with the preservation number of CGMCC 10910.

Wherein, the inoculation amount of the monascus seed liquid in the step 2 is 5-10% of the volume of the basic culture medium.

The preparation method of the monascus seed liquid comprises the following steps: taking the strain to coat the strain in a plate culture medium for culturing for 4-7d, taking bacterial colonies on the plate after culturing, adding the bacterial colonies into a seed culture medium for culturing for 24-36h, and the adding amount of the bacterial colonies in the seed culture medium is 5-10 per 50 mL.

Wherein, the components in the plate culture medium are respectively glucose, potato, agar and water, and the concentration of the glucose is 20g/L, the concentration of the potato is 200g/L and the concentration of the agar is 20 g/L.

Wherein, each component in the seed culture medium is glucose, yeast extract, peptone, KCl and FeSO respectively₄·7H₂O、KH₂PO₄And water, wherein the concentration of glucose is 20g/L, the concentration of yeast extract is 3g/L, the concentration of peptone is 10g/L, the concentration of KCl is 0.5g/L, FeSO₄·7H₂The concentration of O is 0.01g/L, KH₂PO₄The concentration of (2) is 4 g/L.

Wherein, the aerobic fermentation in the step 2 is carried out in a constant temperature oscillating table, the fermentation temperature is 25-35 ℃, and the oscillation frequency is 150-250 rpm.

Wherein the fermentation temperature in the aerobic fermentation in the step 2 is 30 ℃, and the oscillation frequency is 180 rpm.

Compared with the prior art, the invention has the beneficial effects that: according to the invention, the fermentation is carried out in a high-salt extreme environment, the monascus metabolism can be promoted to generate the monascus yellow pigment, so that the yield of the monascus yellow pigment can be obviously improved, and the direct preparation of the culture medium by using seawater or concentrated seawater for fermentation production can be considered, so that the whole production method is low in production cost, environment-friendly and high in yield.

Drawings

FIG. 1 is a graph comparing the yields of extracellular monascus yellow pigment in examples 2 to 4 of the present invention and a control example;

FIG. 2 is a graph comparing the yields of intracellular monascus yellow pigment in examples 2 to 4 of the present invention and in the comparative example;

FIG. 3 is a graph comparing the total yield of monascus yellow pigment in examples 2 to 4 of the present invention and the comparative example.

Detailed Description

The principles and features of this invention are described below in conjunction with the following drawings, which are set forth by way of illustration only and are not intended to limit the scope of the invention.