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Bacillus pumilus mutant and alkaline proteinase produced from the same by fermenting

A technology of bacillus pumilus and protease, applied in the field of bioengineering

Inactive Publication Date: 2008-03-26
BEIJING UNION UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the present invention, the original strain is subjected to ultraviolet mutagenesis to obtain a mutant strain with high production of alkaline protease. At present, there is no report on using this mutant strain to ferment and produce alkaline protease

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Embodiment 1: the influence of different nitrogen sources on producing enzyme

[0014] Add different nitrogen sources to solid medium instead of yeast extract. with 20% Na 2 CO 3 Adjust the pH to 8.5, stir evenly, and sterilize at 115°C for 20 minutes. Inoculate liquid seeds, mix well, and culture statically at 35°C for 48 hours. The results are shown in Table 1. The results showed that the yeast extract was more suitable for the fermentation production of alkaline protease.

[0015] Table 1: Effects of different nitrogen sources on enzyme production

[0016] nitrogen source

Embodiment 2

[0017] Embodiment 2: the impact of different carbon sources added on the production of enzymes

[0018] Various carbon sources were added to the solid medium at 2.5% by weight. with 20% Na 2 CO 3 Adjust the pH to 8.5, stir evenly, and sterilize at 115°C for 20 minutes. Inoculate liquid seeds, mix well, and culture statically at 35°C for 48 hours. The results are shown in Table 1. When D-fructose was added, the enzyme production rate was the highest, reaching 15,300 U / g dry substrate.

[0019] Table 2: Effects of various pectin-containing biomass and medium humidity on enzyme production

[0020] carbon source

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PUM

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Abstract

The present invention provides one kind of Bacillus pumilus mutant with high yield of alkali protease and the process of fermenting with the strain to prepare alkali protease, and belongs to the field of biotechnology. The strain is obtained with Bacillus pumilus strain 1.894 and through ultraviolet mutagenesis and screening. Into wheat bran as main substrate, yeast extract, D-fructose, potassium dihydrogen phosphate, and water are added and Bacillus pumilus strain 2080 is inoculated and cultured statically at air condition and 35 deg.c for 48 hr to reach alkali protease producing peak. Then, the culture is leached with Na2CO3 / NaHCO3 buffering solution and filtered to obtain the filtrate as the alkali protease liquid, and the alkali protease liquid is ultrafiltering concentrated or spray dried to obtain the alkali protease powder. The present invention has alkali protease yield up to 15.3 Kilounit / g dry substrate.

Description

technical field [0001] The invention belongs to the technical field of bioengineering. technical background [0002] Protease is the enzyme with the largest proportion among industrial enzymes, accounting for about 60% of the world's total annual sales. Alkaline protease is widely used in industries such as food, washing and tanning. At present, the main bacterial species producing alkaline protease include bacteria of the genus Bacillus, Streptomyces (such as Streptomyces griseus, Streptomyces fischeri, etc.) and yeast. A Chinese patent (Application No.: 03144206.4) discloses a method for fermenting and producing high-alkaline alkaline protease. The strain used is a strain of basophilic Bacillus subtilis. A Chinese patent (Application No.: 02825897.5) discloses a Bacillus-type alkaline protease from Bacillus species (DSM 14392) and its application in detergents and cleaning agents. A Chinese patent (application number: 00123404.8) discloses a low-temperature alkaline pro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12N9/54C12R1/08
Inventor 李祖明李鸿玉白志辉翟春风朱秋莉
Owner BEIJING UNION UNIVERSITY
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