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Hansenula polymorpha for preparing high-lysine single-cell protein through methyl alcohol and application of Hansenula polymorpha

A technology of Hansenula polymorpha and single-cell protein, which is applied in the field of bioengineering to achieve great social significance and economic value

Active Publication Date: 2016-08-17
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it has not been reported how to use yeast to efficiently utilize methanol to produce single-cell proteins rich in certain amino acids.

Method used

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  • Hansenula polymorpha for preparing high-lysine single-cell protein through methyl alcohol and application of Hansenula polymorpha
  • Hansenula polymorpha for preparing high-lysine single-cell protein through methyl alcohol and application of Hansenula polymorpha

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] This example illustrates the method of performing the first step of plasma mutagenesis to the original strain of Hansenula polymorpha.

[0026] The method for the first step plasma mutagenesis of the original strain of Hansenula polymorpha is as follows:

[0027] The original strain of Hansenula polymorpha was activated and cultured, and the liquid volume in a 100 ml Erlenmeyer flask was 20 ml, shaken at 200rpm in a shaker at 37°C for 24 hours, and a vigorously growing and robust bacterial liquid was obtained; freshly cultured cells were diluted to cell concentration OD 600 =3-4, drop on the sterilized and cooled slide, blow dry with sterile air; use helium as the discharge gas, 100W as the radio frequency power, 10SLM as the gas flow rate, and 10-1800s as the irradiation time Plasma mutagenesis was performed on the bacterial strain, and after the mutagenesis, the bacterial film on the carrier was eluted, and the survival rate was calculated. The experimental results ...

Embodiment 2

[0029] This example illustrates a method for screening Hansenula polymorpha for the efficient use of methanol to produce lysine-rich single-cell proteins.

[0030] Wherein, the culture medium formula used:

[0031] (1) Seed medium: yeast powder 10 g / L, peptone 20 g / L, glucose 20 g / L, and the rest is water.

[0032] (2) Selection medium: methanol 500 mM, ammonium sulfate 1 g / L, potassium dihydrogen phosphate 1 g / L, magnesium sulfate 0.7 g / L, calcium chloride 0.4 g / L, yeast extract 1 g / L, agar The powder is 1.5 g / L, the rest is water, and the pH is natural.

[0033](3) Fermentation medium: 85% phosphoric acid 26.7 ml / L, calcium sulfate dihydrate 0.93 g / L, potassium sulfate 18.2 g / L, magnesium sulfate dihydrate 14.9 g / L, potassium hydroxide 4.13 g / L, glycerin 30 g / L, trace element solution 4 ml / L, water 1 L;

[0034] (4) Fermentation medium 2: 85% phosphoric acid 26.7 ml / L, calcium sulfate dihydrate 0.93 g / L, potassium sulfate 18.2 g / L, magnesium sulfate dihydrate 14.9 g / L, po...

Embodiment 3

[0046] This example illustrates the genetic stability test of Hansenula polymorpha AP23 in Example 2, and the strain subculture and fermentation experiments are as follows.

[0047] The passage stability of the mutant strain AP23 was detected by shake flask fermentation. The results of the subculture fermentation test of bacterial strain AP23 are shown in Table 2:

[0048] Table 2 strain AP23 passage fermentation test result

[0049]

[0050] It can be seen from the table that after 7 passages, the protein and lysine contents of the strain AP23 are stable, and the passaging stability is good.

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Abstract

The invention relates to Hansenula polymorpha for preparing high-lysine single-cell protein through methyl alcohol and application of the Hansenula polymorpha. The classification name of the Hansenula polymorpha is Hansenula polymorpha AP23, the Hansenula polymorpha is preserved in the China Center for Type Culture Collection (CCTCC) on April 5th, 2016, and the preservation number is CCTCC No: M 2016173. By means of the plasma induced mutation technology, the strain capable of producing high-lysine single-cell protein is screened out with methyl alcohol as the only carbon source, and on this basis, the Hansenula polymorpha for efficiently utilizing methyl alcohol is screened out in a gradient mode through the further combination of methyl alcohol. By means of the strain, high-density fermentation can be achieved, methyl alcohol protein production is conducted through a 5 L fermentation tank, the final bacterium wet weight reaches 350-400 g / L, the methyl alcohol protein dry weight reaches 115-130 g / L, the protein content reaches 68%, and the lysine content reaches up to 60 mg / g. The Hansenula polymorpha has great social significance and economic value in development of animal feed protein additives.

Description

technical field [0001] The invention relates to the field of bioengineering, in particular to a Hansenula polymorpha strain using methanol to produce single-cell protein rich in lysine and its application. Background technique [0002] The shortage of protein resources is a major issue facing the world, and all countries are paying attention to strengthening research on the development of new protein resources. Among them, the fastest industrialization is single-cell protein production, which is considered to be one of the most effective ways to solve the shortage of human protein resources. [0003] Single cell protein (single cell protein, SCP) refers to the bacterial protein produced by yeast, mold, fungus, non-pathogenic bacteria and other single-celled microorganisms, and its protein content generally accounts for 40% of the dry matter of the bacteria~ 60%. Methanol protein produced from industrial methanol is called the second-generation single-cell protein. Compared...

Claims

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Application Information

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IPC IPC(8): C12N1/16C12N13/00C12N1/36C12R1/78
CPCC12N1/16C12N1/36C12N13/00C12R2001/78C12N1/165
Inventor 姜岷章文明马江锋董维亮戴仲雪吴若凡
Owner NANJING UNIV OF TECH
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