Method for immobilizing white rot fungi by using bacteria cellulose film as vector

A technology of bacterial cellulose membrane and white rot fungi, applied in the field of applied microorganisms and cell engineering, can solve the problems of large influence of white rot fungi activity, complicated preparation process, low immobilization efficiency, etc., and achieves low environmental impact and biological stability. Strong and good mass transfer performance

Inactive Publication Date: 2009-03-25
NORTHEAST DIANLI UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are still some problems in the actual application of the above carriers, such as the complicated preparation process, low immobilization efficiency, great influence on the activity of white rot fungi, poor operational stability, and easy to cause secondary pollution to the environment after being discarded.

Method used

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  • Method for immobilizing white rot fungi by using bacteria cellulose film as vector
  • Method for immobilizing white rot fungi by using bacteria cellulose film as vector
  • Method for immobilizing white rot fungi by using bacteria cellulose film as vector

Examples

Experimental program
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Effect test

Embodiment 1

[0041] Isolation and identification of white rot fungi and preparation of spore liquid:

[0042] A. Isolation and identification of white rot fungi: Take 2 g of rotten wood and add it to 100 mL of sterilized liquid medium, place it in a biochemical incubator for constant temperature cultivation at 30 ° C, and observe the growth of bacteria in the medium at any time; the liquid culture The percentages of each component in the base to the mass of the liquid medium are: KH 2 PO 4 0.02%, MgSO 4 ·7H 2 O 0.005%, CaCl 2 0.001%, Glucose 1%, NH 3 Cl 0.02%, H 2 o 2 0.05%, 0.028% sodium tartrate, 0.054% sodium succinate, 0.05% potato extract by volume in the liquid culture medium, sterilized at 121°C for 30 minutes; after 48 hours, take 10 mL of the culture solution for repeated enrichment culture, After 3 times of enrichment culture, the final enrichment solution was taken and cultured on a fixed medium plate; the percentages of each component in the solid medium to the mass of...

Embodiment 2

[0045] Preparation of bacterial cellulose membrane:

[0046] A. Isolation and identification of Acetobacter xylinum (Acetobacter xylinum): Dilute vinegar unstrained spirits 100 times and inoculate in 50mL enrichment medium. %; yeast extract 0.5%; sodium acetate 0.2%; CaCO 3 1%; use acetic acid to adjust the pH to 5.0; after sterilizing at 121°C for 30 minutes, add absolute ethanol with a volume percentage of 2% and nystatin with a mass percentage of 0.5%. Cultivate at ℃ for 5 days, and those with milky white colloidal film on the liquid surface are positive. Peel off the gel film on the surface of the enrichment medium, and stick this film carefully on the first separation medium plate; the percentages of each component in the separation medium mass in the separation medium are: glucose 5% ; Peptone 0.5%; Yeast extract 0.5%; Na 2 HPO 4 12H 2 O 0.2%; KH 2 PO 4 0.1%; MgSO 4 ·7H 2 O0.025%; 0.2% citric acid; 2% agar; pH5.8; After the plate was cultured at 28°C for 48 h...

Embodiment 3

[0049] Modification of bacterial cellulose membrane:

[0050] In a 500mL Erlenmeyer flask with stopper, add 20mL of epichlorohydrin, 5mL of absolute ethanol, 1mL of perchloric acid, 3mL of distilled water, dilute to 100mL with toluene, shake well, add 2g of bacterial cellulose obtained in Example 2 The membrane was heated in a constant temperature water bath at 60°C for 4 hours. After the reaction was complete, the membrane was taken out and washed with acetone to remove residual small molecule organic matter on the membrane, and then dried at 80°C. Put the dried film into another 500mL Erlenmeyer flask, add ethylenediamine 6mL, distilled water 100mL, shake fully, heat in 80°C constant temperature water bath for 4 hours, then take out the film and wash it with distilled water until neutral, then After washing with 1 wt% hydrochloric acid, it was still washed with distilled water until neutral, finally rinsed with acetone, and dried naturally to obtain the modified bacterial ce...

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Abstract

The invention discloses a method for immobilizing white-rot fungi by utilizing a bacterial cellulose membrane as a carrier, which includes the following steps: the separation of the white-rot fungi and the preparation of spore solution of the white-rot fungi; the preparation of the bacterial cellulose membrane; the modification treatment of the bacterial cellulose membrane and the preparation of immobilized cells. After the bacterial cellulose membrane is processed by epoxy chloropropane, absolute ethyl alcohol, perchloric acid and the like, the modified bacterial cellulose membrane obtained is adopted as a carrier and reacts with a 25-percent glutaric dialdehyde solution and mixed with the spore solution of the white-rot fungi after the treatment of repetitive washing and the like, and then the solution is sealed and stays overnight at the temperature of 4 DEG C, and repetitively washed to obtain a bacterial cellulose membrane immobilized white-rot fungi. Three continuous batches of treatment tests for organics and chroma of xylitol production wastewater show that the treatment effects of immobilized white-rot fungi are not found to be obviously reduced and the immobilized white-rot fungi has obvious stability.

Description

technical field [0001] The invention belongs to the technical field of applied microorganisms and cell engineering, and relates to a method for preparing an immobilized carrier and a method for using the carrier to immobilize cells. In particular, it relates to a method for immobilizing white-rot fungi by using a bacterial cellulose film as a carrier. Background technique [0002] White rot fungi (White rot fungi) are a kind of saprophytic filamentous fungi. Due to the unique properties of non-specificity and no need for substrate induction, the extracellular lignin-degrading enzymes secreted by them make it resistant to many structures, high toxicity, high Molecular refractory organics have broad-spectrum degradation ability, and have important application value in biopulping and pulp biobleaching, water pollution control and soil remediation in the paper industry. [0003] Although white rot fungi are widely used to degrade various environmental pollutants, there are stil...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/12C12N1/14C12N3/00C12P19/04C12R1/645C12R1/02
Inventor 于大禹张金榜徐富超关晓辉刘文超崔长龙沈凌宏林娟
Owner NORTHEAST DIANLI UNIVERSITY
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