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Accompanying bacterium for increasing fermentation yield of hypocrellin and application thereof

A technology of accompanying bacteria and hypocrellin, which is applied in the direction of bacteria, fermentation, microorganisms, etc., can solve the problems of no large-scale production application, no active ingredients in crude extracts, and indeterminate active ingredients, so as to achieve no pollution to the environment Effect

Active Publication Date: 2017-08-29
SUZHOU UNIV
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  • Abstract
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Problems solved by technology

However, the yield of hypocretin produced by artificial fermentation is relatively low. In order to increase the yield of hypocrellin, common methods include artificial cultivation of fruiting bodies, chemical synthesis, breeding of high-yield strains, optimization of culture conditions, and addition of elicitors, etc.
However, there are few studies on increasing the production of hypocrellin by exogenous induction.
For example, the methanol / chloroform extract of endophytic fungi isolated by Du et al. from bamboo can stimulate and increase the production of hypocrellin, but the active ingredients of the crude extract have not been reported, so the active ingredients that play a stimulating role cannot be determined ( Du W, Liang Z, Zou X, et al. Effects of microbial elicitor on production of hypocrellin by Shiraia bambusicola . Folia microbiologica, 2013, 1-7.)
In addition, the key gene in the synthesis process of hypocretin was transferred in bamboo yellow fungus to increase its content. For example, Deng et al. overexpressed the key enzyme gene multicopper oxidase (MCO) in bamboo yellow fungus , this method has a significant effect on increasing the yield of hypocrellin (Deng H, Gao R, Chen J, et al. An efficient polyethylene glycol-mediatedtransformation system of lentiviral vector in Shiraia bambusicola . ProcessBiochemistry, 2016, 1357-1362.), but because genetic engineering is currently in the experimental stage, there is no application for large-scale production

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  • Accompanying bacterium for increasing fermentation yield of hypocrellin and application thereof

Examples

Experimental program
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Embodiment 1

[0027] In the present embodiment, an associated bacterial strain that improves the fermentation yield of hypocrellin is adopted, and its classification name is: Pseudomonas chrysogenum ( Pseudomonas fulva ), its deposit number is CGMCC No. 13931, and its deposit date is March 27, 2017.

[0028] Pseudomonas chrysogenum ( Pseudomonas fulva ) for the morphological characteristics see the appendix figure 1 shown.

[0029] In this example, LB medium: tryptone 10 g / L, yeast powder 5 g / L, NaCl 10 g / L, pH 7.2-7.4. PDB medium: potato 200 g / L, glucose 20 g / L, yeast extract 5 g / L, KH 2 PO 4 3g / L, MgSO 4 0.75 g / L, thiamine hydrochloride 0.01 g / L.

[0030] Take a single colony of Pseudomonas chrysogenum of the present invention and place it in LB medium, culture it on a shaking table at 30°C for 8 hours, take 1 mL of seed solution and culture it in LB medium under the same conditions for 12 hours, and centrifuge at 4000 rpm 10min, remove the supernatant, add sterile water to ad...

Embodiment 2

[0036] In this example, LB medium: tryptone 10 g / L, yeast powder 5 g / L, NaCl 10 g / L, pH 7.2-7.4. PDB medium: potato 200 g / L, glucose 20 g / L, yeast extract 5 g / L, KH 2 PO 4 3g / L, MgSO 4 0.75 g / L Thiamine hydrochloride 0.01 g / L.

[0037] Take a single colony of Pseudomonas chrysogenum of the present invention and place it in LB medium, culture it on a shaking table at 30°C for 8 hours, take 1 mL of seed solution and culture it in LB medium under the same conditions for 12 hours, and centrifuge at 4000 rpm 10min, remove the supernatant, add sterile water to adjust the concentration of the bacterial solution to 10 4 / mL.

[0038] Take a small piece of bamboo yellow fungus (3 × 3 mm) and place it in PDB medium, culture it on a shaker at 28 °C for 6 days, and take 1 mL of bacterial seed solution (10 4 / mL) was added, and the culture was continued until 8 d to collect the hyphae of bamboo yellow fungus, weigh them, and extract and detect the content of hypocrellin.

[0039] T...

Embodiment 3

[0043] In this example, LB medium: tryptone 10 g / L, yeast powder 5 g / L, NaCl 10 g / L, pH 7.2-7.4. PDB medium: potato 200 g / L, glucose 20 g / L, yeast extract 5 g / L, KH 2 PO 4 3g / L, MgSO 4 0.75 g / L Thiamine hydrochloride 0.01 g / L.

[0044] Take a single colony of Pseudomonas chrysogenum of the present invention and place it in LB medium, culture it on a shaking table at 30°C for 8 hours, take 1 mL of seed solution and culture it in LB medium under the same conditions for 12 hours, and centrifuge at 4000 rpm 10min, remove the supernatant, add sterile water to adjust the concentration of bacteria solution 10 5 / mL.

[0045] Take a small piece of bamboo yellow fungus (3 × 3 mm) and place it in PDB medium, culture it on a shaker at 28 °C for 6 days, and take 1 mL of bacterial seed solution (10 5 / mL) was added, and the culture was continued until 8 d to collect the hyphae of bamboo yellow fungus, weigh them, and extract and detect the content of hypocrellin.

[0046] The same ...

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Abstract

The invention discloses an accompanying bacterium for increasing fermentation yield of hypocrellin and application thereof; the accompanying bacterium has classification name of Pseudomonas fulva, which is collected under CGMCC No. 13931. Viable ones of the accompanying bacteria having different concentrations are added into Shiraia bambusicola fermentation liquid cultured for 5-7 days, and culturing is continued to obtain hypocrellin. The yield of the hypocrellin can be increased effectively, and the accompanying bacterium is nontoxic and harmless to human and livestock and is free of environmental pollution.

Description

technical field [0001] The invention relates to a biological strain, in particular to a natural medicinal fungus sporocarp associated bacterium-Pseudomonas chrysogenum, and also relates to the application of the strain in increasing the production of hypocrellin. Background technique [0002] bamboo yellow fungus ( Shiraia bambusicola P. Heen), also known as Tianzhuhua, Zhusanqi, etc., belongs to the subphylum Ascomycota ( Ascomycotina ), Hypocreataceae ( Hypocreaceae ), Phyllostachys ( Sharaia ). Bamboo yellow fungus mainly parasitizes on the genus Brachyphyllum ( Brachystachyum Keng .) On the delicate branches of bamboo, pink, cracked and tumor-like fruiting bodies are formed, called bamboo yellow, which is mainly distributed in Zhejiang, Yunnan and other places in southern China. As a traditional rare medicinal fungus in my country, it has the ability to relieve cough and relieve Phlegm, expelling wind and dampness, etc., are often used to treat stomach pain, scia...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P15/00C12R1/38
CPCC12P15/00C12N1/205C12R2001/38
Inventor 王剑文马燕军田浩
Owner SUZHOU UNIV
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