Culture medium and fermentation method for producing alginate lyase by fermentation of microvesicle bacterial genus

A technology of alginate lyase and fermentation medium, applied in the field of fermentation, can solve the problems of alginate lyase fermentation technology and fermentation kinetics, lack of brown algae and alginic acid, etc.

Inactive Publication Date: 2015-05-27
JIMEI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] At present, my country's research on alginate lyase is relatively basic, and research at home and abroad is mainly focused on the isolation of strains, the construction of genetically engineered bacteria, the improvement of enzymatic properties, and the structure and performance of enzymatic hydrolysis products. There are few reports on the fermentation process and fermentation kinetics of lyase, and the lack of industrial enzymes for alginate lyase has become a bottleneck for the high-value processing of brown algae and alginic acid.

Method used

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  • Culture medium and fermentation method for producing alginate lyase by fermentation of microvesicle bacterial genus
  • Culture medium and fermentation method for producing alginate lyase by fermentation of microvesicle bacterial genus
  • Culture medium and fermentation method for producing alginate lyase by fermentation of microvesicle bacterial genus

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Experimental program
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Effect test

Embodiment 1

[0029] Example 1: Screening of Microvesicles ALW1

[0030] The present invention first isolates and obtains a strain of Microvesicles ( Microbulbifer sp.ALW1) (CICC deposit number: 23821), and the culture medium of the present invention can effectively increase the unit yield of alginate lyase produced by fermentation of Microvesicles. The kelp samples were added to shake flasks for culture, and the culture medium was gradiently diluted and coated on a plate. A single colony was picked and inserted into the fermentation medium with sodium alginate as the only carbon source. strains such as figure 1 shown. It was identified as the genus Microvesicle and named Microvesicle ALW1.

Embodiment 2

[0031] Example 2: Production of Alginate Lyase by Shake Flask Fermentation of Microvesicles ALW1

[0032] (1) Thaw the glycerol tube strains stored at -20°C and inoculate them in a 250mL shake flask filled with 50mL seed medium, and culture them at 25°C and 180r / min for 48h to obtain shake flask seed liquid. The seed medium is: sodium alginate 5g / L, peptone 5g / L, yeast powder 1g / L, NaCl 30 g / L, MgSO 4 ·7H 2 O 1g / L, K 2 HPO 4 2 g / L, FeSO 4 ·7H 2 O 0.01 g / L, (NH 4 ) 2 SO 4 5g / L, pH 7.5, sterilized at 121°C for 20 minutes;

[0033] (2) Put the cultivated shake flask seed solution into a 250 mL shake flask with 50 mL of fermentation medium at an inoculation amount of 5%, and ferment at 25°C and 180 r / min for 60 h to obtain alginate-containing lyase of fermentation broth. The fermentation medium is: sodium alginate 1g / L, NaCl 60 g / L, K 2 HPO4 3g / L, MgSO 4 ·7H 2 O 0.6g / L, FeSO 4 ·7H 2 O 0.06 g / L, pH 7.5, sterilized at 121°C for 20 min

[0034] (3) Evenly take 1.0 m...

Embodiment 3

[0035] Example 3: Production of alginate lyase by tank fermentation of Microvesicles ALW1

[0036] (1) Thaw the glycerol tube strains stored at -20°C and inoculate them in a 250mL shake flask filled with 50mL seed medium, and culture them at 25°C and 180r / min for 48h to obtain the shake flask seed solution. The seed medium is: sodium alginate 5g / L, peptone 5g / L, yeast powder 1g / L, NaCl 30 g / L, MgSO 4 ·7H 2 O 1g / L, K 2 HPO 4 2 g / L, FeSO 4 ·7H 2 O 0.01 g / L, (NH 4 ) 2 SO 4 5g / L, pH 7.5, sterilized at 121°C for 20 minutes;

[0037] (2) Put the cultured shake flask seed liquid into a 5 L fermenter with 4 L fermentation medium according to the inoculum amount of 5%, and ferment at 25 °C and 100 r / min for 72 h to obtain alginate-containing alginate lyase fermentation broth. The fermentation medium is: sodium alginate 1g / L, NaCl 60 g / L, K 2 HPO 4 3g / L, MgSO 4 ·7H 2 O 0.6g / L, FeSO 4 ·7H 2 O 0.06 g / L, pH 7.5, sterilized at 121°C for 20 minutes, pH 7.5, sterilized at 12...

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Abstract

The invention discloses a culture medium for producing alginate lyase by fermentation of a microvesicle bacterial genus. The culture medium comprises a strain culture medium and a fermentation culture medium. The invention further discloses a fermentation method for producing alginate lyase by the fermentation of microvesicle bacterial genus. According to the fermentation method, on the basis of shake flask fermentation, the screened microvesicle bacterial genus is subjected to fermentation culture with a fermentation tank, a research on fermentation condition optimization is conducted, and a lyase producing rule of the microvesicle bacterial genus in the fermentation tank is researched, so that the yield of alginate lyase is increased effectively; and alginate lyase produced by the fermentation of the microvesicle bacterial genus ALW1 is subjected to pilot scale amplification fermentation, so that important technical parameter guide is provided for future mass production.

Description

technical field [0001] The invention relates to the technical field of fermentation, in particular to a medium for fermenting and producing alginate lyase and a fermentation method thereof. Background technique [0002] Alginate is a high molecular linear polymer composed of α-L-guluronic acid and its C5 epimer β-D-mannuronic acid. The application of alginate is limited due to its large relative molecular weight and low solubility. In recent years, with the deepening of research on oligosaccharides, its high biological activity has gradually attracted attention. The field shows broad application prospects. [0003] Alginate oligosaccharides can be obtained by physical degradation, chemical degradation, and enzymatic degradation. Among them, enzymatic hydrolysis gradually replaces physical and chemical methods due to its mild reaction conditions, strong controllability, and high yield. Alginate lyase can be used as a tool enzyme To prepare biologically active algin oligosac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/88C12R1/01
CPCC12N9/88
Inventor 肖安风蔡慧农倪辉杨帆杜希萍林艳朱艳冰黄高凌杨秋明李利君杨远帆伍菱
Owner JIMEI UNIV
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