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Monascus strain and application thereof in preparing functional monascus

A technology of Monascus pilosa and Monascus, which is applied in the field of Monascus pilosa, can solve the problems of unstable passage, heavy workload, and poor purpose, and achieve the effect of broad application prospects

Active Publication Date: 2015-06-17
陈福生
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the large workload of traditional screening methods and the lack of purpose, it is difficult to obtain an ideal high-yield MK and no citrinin-producing Monascus strain; Controversial issues such as unstable passage and potential safety hazards

Method used

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  • Monascus strain and application thereof in preparing functional monascus
  • Monascus strain and application thereof in preparing functional monascus
  • Monascus strain and application thereof in preparing functional monascus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] The identification of embodiment 1 Monascus MS-1

[0025] A Monascus strain isolated from Monascus that can produce MK but not citrinin was identified by morphology and ITS sequence analysis.

[0026] 1. Morphological identification of Monascus MS-1

[0027] 1. Medium

[0028] (1) Malt juice agar medium (MA): 1000mL of 10°Bx wort juice, 15g of agar.

[0029] (2) Saccharomyces tsarzia extract agar medium (CYA): NaNO 3 3.0g, K 2 HPO 4 1.0g, KCl0.5g, MgSO 4 ·7H 2 O0.5g, FeSO 4 ·7H 2 O0.01g, yeast extract 5.0g, glucose 20g, agar 15g, add distilled water to 1000mL.

[0030] (3) Glycerol nitrate agar medium (G25N): Add 25% glycerol (w / w) to CYA medium.

[0031] (4) Potato Dextrose Agar (PDA) Medium: Peel 200g of potatoes, cut into small pieces, add 1000mL of distilled water and boil for 20min, after filtering, add 20g of glucose, 15g of agar powder, and dilute to 1000mL with distilled water.

[0032] All the above four media were sterilized by moist heat at 121°C f...

Embodiment 2

[0050] Example 2 Preparation of solid-state fermentation product of Monascus pilosa strain MS-1 and detection of MK and citrinin

[0051] 1. Culture medium

[0052] 1. PDB medium: PDA medium without adding agar powder is PDB medium.

[0053] 2. Yeast extract sucrose (YES) medium: Yeast extract 40g / L, sucrose 160g / L, dilute to 1000mL with distilled water.

[0054] Two, the preparation of Monascus pilosa strain MS-1 seed solution

[0055] Same as the content of the invention.

[0056] 3. Preparation of solid-state fermentation products of Monascus pilosa strain MS-1

[0057] Crush the rice (20 mesh), adjust the water content to 20%, sterilize at 121°C for 20 minutes and break up while hot, inoculate the activated liquid seed solution with 10% (v / w) inoculum and use sterile water Adjust the water content and culture at 28°C for 14 days. The fermented red yeast rice flour was dried at 55°C for 12 hours, and the amount of MK and citrinin was extracted and analyzed.

[0058] 4...

Embodiment 3

[0066] Example 3 Analysis of Synthetic Related Genes of Monascus Placiferus Strain MS-1 Citrinin

[0067] Using molecular biology methods, the citrinin high-yielding strain Monascus ruber (M. ruber) M-7 (CCAM070120) was used as a positive control (Li L., Shao Y.C., Li Q. et al.. Identification of Mga1, a G-protein α-subunit gene involved in regulating citrinin and pigment production in Monascus ruber M-7.FEMS Microbiology Letters.2010,308:108-114.), its citrinin synthesis-related genes (ctnA, ctnE, ctnR, pks CT) analysis, Monascus genome extraction method is the same as ITS sequence analysis, the reaction system is 25 μL, including Monascus genome DNA 1 μL, 10 μmol / L forward and reverse primers 1 μL, dNTP 1 μL, 10×buffer 2.5 μL, 2.5mmol / LTaq plus DNA Polymerase0.2μL, ddH 2 O18.3 μL. The PCR reaction program was pre-denaturation at 95°C for 5min, denaturation at 95°C for 30s, annealing at 55°C for 30s, extension at 72°C for 1min, a total of 30 cycles, and a final extension at...

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Abstract

The invention discloses a monascus strain and application of the monascus strain in preparing functional monascus. The Monascus sp. strain MS-1 provided by the invention is conserved in China Center for Type Culture Collection on June 26, 2013 with the conservation number of CCTCC M2013295. The strain is identified as M.pilosus through morphology and an ITS sequence analysis method. The Monascus sp. strain MS-1 does not contain ctnA, ctnE, ctnR or pksCT genes related to the synthesis of citrinin in the genome and does not have the citrinin synthesizing capability. The invention further provides a method for producing the functional monascus rich in monacolin K but containing no citrinin. The method comprises the following steps: (1) bringing the Monascus sp. strain MS-1 into a seed medium for enrichment culture to obtain a seed solution; (2) taking the seed solution into a fermentation medium for solid fermentation, wherein a temperature change culture method is adopted, the seed solution is cultured for three days at 30 DEG C and then cultured for 11 days at 25 DEG C; and (3) drying the fermentation product at 55 DEG C to obtain the functional monascus. The content of the monacolin K in the functional monascus is high, and the detection shows no citrinin exists.

Description

technical field [0001] The invention belongs to the fields of fermentation engineering and biotechnology, and in particular relates to a monascus pilosus strain with high yield of monacolin K but not citrinin and its application. Background technique [0002] Monascus spp., also known as Monascus spp., is an important edible microbial resource in my country. Its solid-state fermentation product, Monascus spp., which uses rice as a culture substrate, is used as a food colorant and preservative in my country and Southeast Asia. , Fermentation agent and traditional Chinese medicine compatibility, etc. have been used for thousands of years. Studies have shown that Monascus can produce abundant secondary metabolites, such as monacolins, Monascus pigments, γ-aminobutyric acid, dimerumic acid, etc. However, some strains of Monascus may also produce a mycotoxin, citrinin, which may contaminate Monascus products and pose a safety hazard. For this reason, the screening of Monascus st...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C12P1/02C12R1/645
Inventor 陈福生冯艳丽
Owner 陈福生
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