Culture medium combination for promoting natural growth and transformation of bacillus subtilis and transformation method

A Bacillus subtilis, natural growth technology, applied in the field of microorganisms, can solve the problems of low transformation efficiency, complicated operation, difficulty in preparing protoplasts by Bacillus subtilis, etc., and achieves high transformation efficiency, simple and easy method, and the effect of improving complex methods.

Active Publication Date: 2020-02-04
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented innovation allows bacteria called Bacillusthuringulossis (BSC) or other microorganisms to grow naturally without being affected by external factors like light or temperature conditions during their culture process. These cultures are then transformed into useful products such as enzymes from these organism's own genome. By doing this, they have potential applications in various industries including food production, medicine development, biotechnology researchers, etc., which could benefit many people worldwide.

Problems solved by technology

The technical problem addressed in this patented text relates to finding ways to enhance the growth rates or transformation efficiencies of microorganisms such as Bacilli while simplifying their preparation process.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1 A kind of culture medium combination of natural growth transformation of Bacillus subtilis

[0038] Medium SP I: (NH 4 ) SO 4 0.1g; K 2 HPO 4 0.2g; KH 2 PO 4 0.4g; Sodium citrate dihydrate (Na-Citrate-2H 2 O) 0.2g; MgSO 4 ·7H 2 O 0.01g; 20% (mass fraction)*glucose (Glucose) 2.4mL; 1% (mass fraction)*yeast extract (Yeast extract) 9mL; 10mg / mL*L-leucine (L-Leucine) 450μL; 10mg / mL*L-Methionine (L-Methionine) 450μL; 0.2% (mass fraction)*casein hydrolyzate (Casamino acid) 9mL (the above with * should be sterilized separately), with dH 2 O was dissolved and adjusted to 100mL for sterilization.

[0039] Medium SP II: (NH 4 ) SO 4 0.1g; K 2 HPO 4 1.3g; KH 2 PO 4 0.5g; Sodium citrate dihydrate (Na-Citrate-2H 2 O) 0.06g; MgSO 4 ·7H 2 O 0.01g; 20% (mass fraction)*glucose (Glucose) 2.4mL; 1% (mass fraction)*yeast extract (Yeast extract) 1.8mL; 15mg / mL*L-leucine (L-Leucine) 48μL ; 10mg / mL*L-Methionine (L-Methionine) 48μL; 500mmol / L*MgCl

Embodiment 2

[0040] Embodiment 2 A kind of culture medium combination of natural growth transformation of Bacillus subtilis

[0041] Medium SP I: (NH 4 ) SO 4 0.3g; K 2 HPO 4 0.3g; KH 2 PO 4 0.45g; Sodium citrate dihydrate (Na-Citrate-2H 2 O) 0.25g; MgSO 4 ·7H 2 O 0.014g; 20% (mass fraction)*glucose (Glucose) 2.45mL; 1% (mass fraction)*yeast extract (Yeast extract) 10.5mL; 10mg / mL*L-leucine (L-Leucine) 470μL ; 10mg / mL* L-methionine (L-Methionine) 470μL; 0.2% (mass fraction)* casein hydrolyzate (Casaminoacid) 9.6mL (the above with * should be sterilized separately), use dH 2 O was dissolved and adjusted to 100mL for sterilization.

[0042] Medium SP II: (NH 4 ) SO 4 0.21g; K 2 HPO 4 1.36g; KH 2 PO 4 0.55g; Sodium citrate dihydrate (Na-Citrate-2H 2 O) 0.07g; MgSO 4 ·7H 2 O 0.016g; 20% (mass fraction) * glucose (Glucose) 2.35mL; 1% (mass fraction) * yeast extract (Yeast extract) 1.9mL; 15mg / mL * L-leucine (L-Leucine) 49.2 μL; 10mg / mL*L-Methionine (L-Methionine) 4

Embodiment 3

[0043] Embodiment 3 A kind of culture medium combination of natural growth transformation of Bacillus subtilis

[0044] Medium SP I: (NH 4 ) SO 4 0.3g; K 2 HPO 4 0.5g; KH 2 PO 4 0.5g; Sodium citrate dihydrate (Na-Citrate-2H 2 O) 0.26g; MgSO 4 ·7H 2 O 0.021g; 20% (mass fraction)*glucose (Glucose) 2.56mL; 1% (mass fraction)*yeast extract (Yeast extract) 10.2mL; 10mg / mL*L-leucine (L-Leucine) 510μL ; 10mg / mL* L-methionine (L-Methionine) 510μL; 0.2% (mass fraction)* casein hydrolyzate (Casaminoacid) 10mL (the above with * should be sterilized separately), with dH 2 O was dissolved and adjusted to 100mL for sterilization.

[0045] Medium SP II: (NH 4 ) SO 4 0.26g; K 2 HPO 4 1.42g; KH 2 PO 4 0.62g; Sodium citrate dihydrate (Na-Citrate-2H2O) 0.11g; MgSO 4 ·7H 2 O 0.02g; 20% (mass fraction) * Glucose (Glucose) 2.53mL; 1% (mass fraction) * Yeast extract (Yeast extract) 2.2mL; 15mg / mL * L-leucine (L-Leucine) 50μL ; 10mg / mL*L-Methionine (L-Methionine) 50μL; 50

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Abstract

The present invention discloses a culture medium combination for promoting natural growth and transformation of bacillus subtilis and a transformation method. The culture medium combination comprisesa culture medium 1 and a culture medium 2. The culture medium containing a special nutrient formula is utilized to efficiently obtain bacillus subtilis competent cells, the natural transformation method is cooperated and beneficial recons are more efficiently and quickly obtained, thus a complex method that can only rely on electrotransformation of the bacillus subtilis is improved, transformationcost is greatly reduced, and foundation for directed evolution and metabolic engineering of proteins of the bacillus subtilis is laid. The technology overcomes shortcomings of traditional transformation technology, and the method is simple and easy to implement, high in conversion efficiency and easy to popularize.

Description

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Claims

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Application Information

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Owner SOUTH CHINA AGRI UNIV
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