Method for preparing epsi-polylysine and its salt by using Kitasatosporia PL6-3

A technology of Lispora militaris and polylysine, applied in the field of fermentation engineering, can solve the problems of difficult production, low fermentation yield, viscous fermentation liquid, etc., and achieve convenient operation, broad-spectrum bacteriostasis, and extensive culture conditions Effect

Inactive Publication Date: 2005-11-16
NANJING UNIV OF TECH
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  • Abstract
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Problems solved by technology

Since Streptomyces is an actinomycetes, mycelium winding is relatively serious, and the fermentation liquid in the later stage is very viscous, so the

Method used

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  • Method for preparing epsi-polylysine and its salt by using Kitasatosporia PL6-3
  • Method for preparing epsi-polylysine and its salt by using Kitasatosporia PL6-3

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Slant Medium: Glucose 1%, Beef Extract 0.5%, Yeast Extract 0.5%, K 2 HPO 4 0.1%, MgSO 4 .7H 2 O 0.05%, agar 2%, pH7.0

[0069] Shake flask medium: Glucose 5%, (NH 4 ) 2 SO 4 1%, Na 2 HPO 4 .12H 2 O 0.158%, KH 2 PO 4 .7H 2 O 0.136%, MgSO 4 .7H 2 O 0.05%, ZnSO 4 .7H 2 O 0.004%, FeSO 4 .7H 2 O0.003%, yeast extract 0.5%, pH6.8. Fill 50ml of liquid in the Erlenmeyer flask of 250ml capacity, 121 ℃ sterilize 15 minutes.

[0070] Cultivate the purified Lispora militaris PL6-3 (CCTCC No.M205012) on a slant medium at 28-32°C for 72 hours, then put a ring of spores of this bacteria in the shake flask medium, culture at 30°C for 72 hours, and shake the flask at a rotating speed of 72 hours. 200r / min. Finally, the content of ε-polylysine in the fermented liquid was 0.944g / L.

Embodiment 2

[0072] Glucose 150g, yeast extract 15g, (NH 4 ) 2 SO 4 30g, MgSO 4 .7H 2 O 0.75g, FeSO 4 .7H 2 O 0.09g, ZnSO 4 .7H 2 O 0.12g, KH 2 PO 4 .7H 2 O 4.08g, Na 2 HPO 4 .12H 2 O 4.74g, adjust the pH to 6.8 with NaOH, adjust the volume to 3L with water, put it into a 5L glass stirred fermenter, and steam sterilize at 121°C for 15 minutes. K.sp.PL6-3 (CCTCC No.M205012) was cultured at 30° C. for 24 hours with a seed medium, which was the same as the fermentation medium. Insert 300ml of the seed liquid into the fermented liquid after cooling, cultivate at 30°C, (ventilation ratio 1: 2.5vvm, stirring speed is 350r / min), and as the cultivation time prolongs, the ε-polylysine in the culture liquid will continue to increased, and at 72 hours, the fermentation broth contained ε-polylysine 3.0g / L.

Embodiment 3

[0074] Same as in Example 2, the fermentation process is controlled at pH 4.0, glucose and nitrogen sources are added to maintain the glucose concentration at about 10 g / L, and after 168 hours of fermentation, 25 g / L of ε-polylysine is accumulated in the fermentation broth. Centrifuge the fermentation broth to remove bacteria, adsorb on 732 cation exchange resin on the supernatant, wash with water, elute with 1N ammonia water, concentrate the collected liquid to remove ammonia, decolorize with activated carbon, continue to concentrate and then precipitate with ethanol to obtain ε-polylysine hydrochloride Salt 15.25g. The specific rotation of the product α D 25 =+57.1°(c,1H 2 0), the molecular weight measured by SDS-PAGE electrophoresis is 5000Da.

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Abstract

A process for preparing epsilon-polylysine and its salt from kitasatospora sp. PL6-3 (CCTCC No.M205012) includes screening kitasatospora sp.PL6-3, culturing in the culture medium containing carbon and nitrogen sources, fermenting, centrifugal separation or filtering for removing thallus, and separating epsilon-polylysine and its salt by ion exchange resin method.

Description

technical field [0001] The invention belongs to the technical field of fermentation engineering, in particular to a microbial strain Kitasatospora sp. PL6-3 with high yield of ε-polylysine (ε-PL), and its use in ε-polylysine and its salt production. Background technique [0002] ε-poly-L-lysine (ε-poly-L-lysine, ε-PL) is a homopolymer of L-lysine synthesized by microorganisms, which is formed by linking ε-amino and α-carboxyl groups through peptide bonds. ε-polylysine has the activity of resisting Gram-positive bacteria, Gram-negative bacteria, fungi and viruses, and is a biological preservative with excellent performance. In addition, it is also used as cosmetics, gene carriers, drug coatings, electronic materials and environmental protection materials. [0003] In 1977, when Sakai Heiichi and Shimao Shoji screened a large number of D.P. positive substances (alkaloids), they found that there was a large amount of D.P. substances in the fermentation broth of Streptomyces a...

Claims

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Application Information

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IPC IPC(8): C12N1/00
Inventor 徐虹王军姚忠欧阳平凯
Owner NANJING UNIV OF TECH
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