Serratia marcescens Sm-128 strain with high prodigiosin production and its application

A technology of Serratia marcescens and sm-128 is applied in the fields of biotechnology medicine and life, and can solve the problems of lack of high-yielding prodigiosin strains, imperfect fermentation production process and the like

Inactive Publication Date: 2011-12-14
ZHEJIANG NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Aiming at the increasing demand for prodigiosin in the market, the lack of high-yielding prodigiosin strains and the imperfect fermentation process

Method used

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  • Serratia marcescens Sm-128 strain with high prodigiosin production and its application
  • Serratia marcescens Sm-128 strain with high prodigiosin production and its application
  • Serratia marcescens Sm-128 strain with high prodigiosin production and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 obtained from the isolation and screening of freshwater fish Sm-128 strain

[0028] 1.1 Medium

[0029] PDA medium: potato 200 g / L, glucose 20 g / L, agar 20 g / L, natural pH.

[0030] LB medium: tryptone 10 g / L, yeast extract 5 g / L, NaCl 10 g / L, agar 20 g / L, pH7.0-7.2.

[0031] Isolation, screening and identification of strains

[0032] Take a freshwater crucian carp, soak and wash it with sterile water, and dissect the fish body to obtain surface scales, muscle tissue and viscera. Soak in sterile water at low temperature overnight under airtight conditions, and adopt the concentration gradient dilution method (10 -1 -10 -4 ), spread 200 μL of the diluted solution on a PDA medium plate, and incubate at a constant temperature of 28 °C. A single colony (1 day) with white early colonies and red or purple-red colonies was picked and separated and purified by three consecutive streaks to obtain the Sm-128 strain with a single character. Use LB solid sla...

Embodiment 2

[0037] Embodiment 2 The method that Serratia marcescens Sm-128 strain is used for the synthesis of prodigiosin

[0038] 2.1 Medium

[0039] LB medium: tryptone 10 g / L, yeast extract 5 g / L, NaCl 10 g / L, agar 20 g / L, pH7.0-7.2.

[0040] Fermentation medium: peptone 9 g / L, glycerol 10 ml / L, pH 7.4.

[0041] Biosynthesis of prodigiosin

[0042]Pick about 2 rings of strains from the activated LB medium and insert them into the culture medium, cultivate them at 28 °C and 150 r / min for 12 h, then insert them into the fermentation medium with an inoculum of 3 %, at 28 °C, 150 r / min / min and cultured for 24 h, the Serratia marcescens cell containing prodigiosin can be obtained, and a small amount of the target product is in the fermentation broth.

[0043] Determination of medium and fermentation conditions

[0044] Using LB medium as the starting medium, the medium was optimized, and the carbon source was finally determined to be glycerol, the nitrogen source was peptone, ...

Embodiment 3

[0045] Example 3 Extraction method of prodigiosin produced by Serratia marcescens Sm-128 strain

[0046] After the fermentation broth was centrifuged and washed, the bacterial cells were obtained, and 30% bacterial suspension was configured with absolute ethanol. After crushing by high-pressure homogenization, it was repeatedly extracted with ethyl acetate until the bacteria were colorless. The ethyl acetate phases were combined and concentrated to obtain a crude pigment. The crude product was redissolved with ethanol, and TLC experiment was carried out to obtain the best developer n-hexane:ethyl acetate=1:40. Using this developer as the eluent, perform column chromatography with 200 mesh silica gel to obtain an orange component a1, evaporate a1 to dryness and redissolve it in methanol, and pass through a LH-20 gel column to obtain three components b1, b2, and b3. Among them, the b2 component is the dominant component. After drying, it is analyzed by ultraviolet absorption...

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Abstract

The invention belongs to the field of biotechnological medicine and life and particularly relates to a high-prodigiosin-yield serratia marcescens strain Sm-128 and use thereof. The high-prodigiosin-yield serratia marcescens strain Sm-128 was collected in China Center for Type Culture Collection on November 13, 2010. The collection number of the strain is CCTCC No.M2010347. The serratia marcescens strain Sm-128 can be used for synthesizing prodigiosin. According to ultraviolet absorption spectrum and Fourier H nuclear magnetic resonance (H-NMR) spectrum analysis, the pigment produced by the Sm-128 strain is prodigiosin and the yield of the prodigiosin reaches 3 to 5 g / L. The pigment has high thermostability, oxidant resistance and reducer resistance and low photostability.

Description

technical field [0001] The invention belongs to the fields of biotechnology, medicine and life, and in particular relates to a strain of Serratia marcescens with high prodigiosin production ( Serratia marcescens ) Sm-128 strain and its screening method and application. Background technique [0002] Prodigiosins (PG) is a general term for a class of natural red pigments with a tripyrrole ring, which have antibacterial, antifungal, antimalarial, immunosuppressive and antitumor activities. In recent years, research on prodigiosin has mainly focused on its potential clinical application. Prodigiosin is a promising antitumor drug with an average IC50 (half inhibitory concentration) of 2.1 μmol / L. It has strong resistance to 57 different cancer cells in the human body, and is also resistant to various human cancer cells such as liver cancer metastasis in vitro and human primary cancer cells, and has no obvious toxicity to non-malignant cell lines. [0003] As a secondary metabol...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P17/16C12R1/43
Inventor 蒋冬花韦凤蔡琪敏蓝丽精嵇豪高爱同
Owner ZHEJIANG NORMAL UNIVERSITY
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