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Immobilization method for glossy ganoderma cell

A technology for immobilizing cells and cells, which is applied in the direction of immobilization on/in organic carriers, can solve the problems of low product content and yield, low cell density, etc., and achieve easy processing, high immobilization efficiency, and immobilization process simple effect

Active Publication Date: 2011-06-08
宋秋兰
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, liquid submerged fermentation uses free cells to produce ganoderma sugar and ganoderma acid. The cell density is low, and the product content and yield are low. This is the bottleneck of the ganoderma polysaccharide and ganoderma acid production industry.

Method used

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  • Immobilization method for glossy ganoderma cell
  • Immobilization method for glossy ganoderma cell
  • Immobilization method for glossy ganoderma cell

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] (1) Preparation of artificial sponge immobilized cell carrier: take an artificial sponge with a pore size of 0.75mm, wash it, and cut it into 0.3cm 3 , sterilized at 121°C for 50 minutes, and stored for later use.

[0024] (2) Slant culture of Ganoderma lucidum cells: Under sterile conditions, inoculate Ganoderma Sinense (Ganoderma Sinense) YTZ9k1 CGMCCNo.0742 cells on the slant medium, culture at 25-28°C for 8 days, and form seed bacteria for later use. Among them, the slant medium contains potato 200g / L, agar 20g / L, peptone 0.2g / L, glucose 20g / L, potassium dihydrogen phosphate 0.3g / L, magnesium sulfate 0.15g / L, vitamin B 1 0.08g / L.

[0025] (3) Expanded culture of seed bacteria: under sterile conditions, collect seed bacteria, inoculate into a 500ml Erlenmeyer flask filled with 300ml of sterile culture solution, and cultivate at 100-130r / min shake flask for 8-10 days at 25-28°C. form bacteria. Among them, the culture solution of seed bacteria contains soybean 45g / L...

Embodiment 2

[0033] (1) Preparation of corn cob immobilized cell carrier: take mature corn cobs, remove corn kernels, cut off about 0.5-1 cm from both ends of corn cobs, wash, sterilize at 121°C for 50 minutes, and store for later use.

[0034] (2) Slant culture of Ganoderma lucidum cells: Under sterile conditions, inoculate Ganoderma Sinense (Ganoderma Sinense) YTZ9k1 CGMCCNo.0742 cells on the slant medium, culture at 25-28°C for 8 days, and form seed bacteria for later use. Among them, the slant medium contains potato 200g / L, agar 20g / L, peptone 0.2g / L, glucose 20g / L, potassium dihydrogen phosphate 0.3g / L, magnesium sulfate 0.15g / L, vitamin B 1 0.08g / L.

[0035](3) Expanded cultivation of seed bacteria: under aseptic conditions, collect seed bacteria, inoculate into a 2000ml Erlenmeyer flask containing 1200ml of sterile culture solution at 25-28°C, and shake the flask at 100-130r / min for 8-10 days to form bacteria liquid. Among them, the culture solution of seed bacteria contains soybe...

Embodiment 3

[0043] (1) Preparation of immobilized cell carrier of loofah sponge body: dry mature loofah gourd and remove the skin part to obtain loofah sponge body, cut into 0.3cm 3 , washed, sterilized at 121°C for 50 minutes, and stored for later use.

[0044] (2) Slant culture of Ganoderma lucidum cells: Under sterile conditions, inoculate Ganoderma Sinense (Ganoderma Sinense) YTZ9k1 CGMCCNo.0742 cells on the slant medium, culture at 25-28°C for 8 days, and form seed bacteria for later use. Among them, the slant medium contains potato 200g / L, agar 20g / L, peptone 0.2g / L, glucose 20g / L, potassium dihydrogen phosphate 0.3g / L, magnesium sulfate 0.15g / L, vitamin B 1 0.08g / L.

[0045] (3) Expanded cultivation of seed bacteria: under sterile conditions, collect seed bacteria, inoculate into a 2000ml triangular flask (expanded cultivation) filled with 1200ml of sterile culture solution, and culture at 100~130r / min for 8~ After 10 days, a bacterial liquid was formed. Among them, the culture ...

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Abstract

The invention discloses a method for immobilizing glossy ganoderma cells, comprising the following steps: (1) the preparation of carriers of immobilizing cells: artificial sponges, corncobs or vegetable sponges are cleaned, sterilized for 45 to 60 min under the temperature of 121 DEG C, and preserved in sterile condition; (the following steps are carried out in sterile condition) (2) the cultivation of glossy ganoderma cells on an inclined medium: the glossy ganoderma cells are inoculated on an inclined medium under the temperature of 25 to 28 DEG C, and after 7 to 9 days, seed strains are formed; (3) further cultivation of the seed strains: the seed strains are collected and then inoculated into a 500ml-volume flask with 250-350ml culture medium or a 2,000ml-volume flask with 1100-1400mlculture medium under the temperature of 25 to 28 DEG C, and after 8 to 10 days of cultivation by shaking the flask at a speed of 100-130r / min the strain medium is made; (4) the dry weight of the cells measured by weighing method is 0.7 to 1.0mg / ml; (5) standby carriers are added according to the ratio of the cultivation medium volume to the dry weight of the carrier of 8-10ml to 1-1.5mg; and (6) the cells are immobilized after 8 to 10 hours of cultivation by shaking the flask at a speed of 100-130r / min.

Description

technical field [0001] The invention relates to a microbial cell immobilization method, in particular to a ganoderma cell immobilization method. Background technique [0002] Ganoderma lucidum (Leyss ex Fr.) Krast.) belongs to Basidiomycetes, Polyporaceae, Ganoderma higher fungi. Traditional Chinese medicine believes that Ganoderma lucidum is sweet in taste, warm in nature, non-toxic, and can replenish the five swollen qi of the heart, liver, lung, spleen, and kidney. Modern medical research also shows that the pharmacological components of Ganoderma lucidum are very rich, and the active ingredients include polysaccharides (namely: Ganoderma lucidum polysaccharides), triterpenoids (mainly Ganoderma acids), Ganoderma lucidum polypeptides, 16 kinds of amino acids (including seven kinds of human essential amino acids), proteins, steroids, mannitol, coumarin, alkaloids, organic acids (mainly containing fumaric acid), and trace elements Ge, P, Fe, Ca, Mn, Zn, etc. Ganoderma luc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N11/02
Inventor 宋秋兰
Owner 宋秋兰
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