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Penicillium camemberti and application thereof

A technology of salmonella cheese and penicillium, applied in the field of medicine, can solve the problems of low conversion rate, achieve low production cost, high concentration of conversion products, and avoid discharge and treatment

Inactive Publication Date: 2013-07-31
SHENYANG PHARMA UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far, most of the reports use cis-acrylphosphate as a substrate for biotransformation (as shown in Table 1), and in the past two years, it has been reported that D-fosfomycin is a substrate for biotransformation research, but the conversion rate Very low (only 5.41%), the concentration of the transformant is 0.27mg / ml【Zhou Lisha; An Haiying; Li Hui; Yang Weichao; Research. Biotechnology Bulletin, 2008 Supplement, 374-377.]

Method used

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  • Penicillium camemberti and application thereof
  • Penicillium camemberti and application thereof
  • Penicillium camemberti and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment one, the preparation of fosfomycin

[0033] 1. Penicillium camemberti was cultured on a PDA agar slant at 28° C. for 24 hours.

[0034] 2. Seed culture medium: glucose 10.0g, distill potato juice to 1L. pH6.8, sterilized at 121°C for 20min. Potato dipping juice: 200.0g of peeled potatoes, add 1L of distilled water and boil for 30 minutes, filter with four layers of gauze to get the dipping juice. Pack into 250ml Erlenmeyer flasks, each 30ml.

[0035] 3. Transformation medium: 3.0 g of dexfosfomycin double salt, dilute the potato juice to 1 L. Natural pH (pH5.7), sterilized at 121°C for 20min. Potato dipping juice: 200.0g of peeled potatoes, add 1L of distilled water and boil for 30 minutes, filter with four layers of gauze to get the dipping juice. Pack into 250ml Erlenmeyer flasks, each 60ml.

[0036] 4. Inoculate the strain in step 1 according to the inoculum size of 1.62×10 8 1 spore / bottle was inoculated into the seed culture medium in step 2, at 2...

Embodiment 2

[0038] Embodiment two, the detection and identification of fosfomycin

[0039] 1. Indicator bacteria culture and suspension preparation

[0040] Indicator bacteria: Micrococcus luteus

[0041] Medium: peptone 10.0g, yeast powder 5.0g, NaCl 10.0g, agar 20.0g, distilled water to 1L. pH7.2~7.4, after sterilizing at 121℃ for 20min, place on an inclined plane for later use.

[0042] Cultivation of indicator bacteria: cultured at 37°C for 24 hours, the bacterial lawn turns yellow.

[0043]Preparation of indicator bacteria suspension: take a slope with good growth condition, add sterile normal saline, gently scrape off the bacterial lawn with a bamboo stick, wait for OD 600 After reaching 0.2-0.3, transfer to a sterile empty bottle and store at 4°C for later use. 2. Preparation of biological assay medium and its identification plate

[0044] Bioassay medium: peptone 10.0g, yeast powder 5.0g, NaCl 10.0g, agar 20.0g, distilled water to 1L. pH 7.2-7.4, sterilized at 121°C for 20 m...

Embodiment 3

[0049] Embodiment three, the factor that influences transformation

[0050] 1. Effect of feeding time on biotransformation

[0051] Feeding time is one of the most important factors affecting biotransformation. Table 2 shows the relationship between feeding time and conversion rate when the substrate concentration is 3.0 mg / mL.

[0052] Table 2 The relationship between different feeding time and conversion

[0053]

[0054] 2. The effect of substrate dosage on biotransformation

[0055] Substrate concentration is also one of the important factors affecting biotransformation. Table 3 shows the relationship between substrate concentration and transformation.

[0056] Table 3 Relationship between different substrate concentrations and conversion

[0057]

[0058] 3. The impact of conversion time on conversion

[0059] It is very important to determine the appropriate conversion end point to improve the production capacity and economic benefit of the product. Table 4 sh...

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Abstract

The invention provides penicillium camemberti (Penicillium camemberti). The strain has been preserved in China General Microbiological Culture Collection Center (CGMCC) on June 2, 2010, and the preservation number is CGMCC No.3904. The strain has the characteristics that: the strain grows well on a PDA inclined plane; the bacterial colony diameter on the PDA is about 28mm; the bacterial colony isgrayish green and has short villiform texture and protruding center; the aerial mycelium is white without wrinkle; the mycelium inside the matrix is faint yellow without spillage; the bacterial colony does not grow at the temperature of between 5 and 37 DEG C; the conidium has smooth peduncle, stalk length of between 100 and 400 mu m, expanded top end, and the diameter of about 2-4 mu m; the sporiparous structure is in multiple whorls, and has ampoule bottle-shaped stalks with the length of 7-10 mu m*2-2.5mu m; and the conidium is catenate, elliptical and smooth and has the diameter of 3.0 to3.5 mu m. The invention also provides a method for preparing medicinal levophosphonomycin by bioconverting dextrophosphonomycin through microbes, which comprises the steps of: culturing the penicillium camemberti, and separating the medicinal levophosphonomycin from the culture supernatant.

Description

Technical field: [0001] The invention belongs to the technical field of medicine, and relates to a Penicillium camemberti and its application, in particular to a Penicillium camemberti (Penicillium camemberti) and its preparation of levofosfomycin by biotransformation of Penicillium method of fosfomycin. Background technique: [0002] Fosfomycin [(-)-(1R,2S)-1,2-epoxypropylphosphonic acid, (-)-(1R,2S)-1,2-epoxypropylphosphonic acid, fosfomycin, phosphonomycin, FOM] is a bacterial An important clinically important broad-spectrum antibiotic for cell wall synthesis, discovered in 1969 by Hendlin et al. from Streptomyces freundii. It covalently binds to pyruvyltransferase, causing irreversible inactivation of the enzyme and inhibiting the synthesis of N-acetylmuramic acid. Fosfomycin has no cross-resistance with other antibiotics, so it can be used clinically for infections caused by drug-resistant Staphylococcus aureus, Escherichia coli, Proteus, Pseudomonas aeruginosa and Sa...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C12P17/02C12R1/80
Inventor 张怡轩张景海吴春福
Owner SHENYANG PHARMA UNIVERSITY