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High-yield heat resistant type neutral phytase bacterial strain as well as fermentation culture medium and enzyme production method thereof

A fermentation medium and phytase technology, applied in the field of microorganisms, can solve the problems of no heat resistance, inability to resist enzyme inactivation, increase costs, etc., and achieve the effect of improved ability

Inactive Publication Date: 2012-04-25
ZHEJIANG GONGSHANG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Secondly, the heat resistance of phytase is generally low. Without good heat resistance, it cannot resist the enzyme inactivation caused by high-temperature pelleting of feed, which further increases its cost.

Method used

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  • High-yield heat resistant type neutral phytase bacterial strain as well as fermentation culture medium and enzyme production method thereof
  • High-yield heat resistant type neutral phytase bacterial strain as well as fermentation culture medium and enzyme production method thereof
  • High-yield heat resistant type neutral phytase bacterial strain as well as fermentation culture medium and enzyme production method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1. Screening of high-yielding heat-resistant neutral phytase strains

[0024] Take samples from the rhizosphere soil of plants in Zhejiang and Yunnan, and soil samples from high-temperature zones. After bathing in 80°C water for 10 minutes, common miscellaneous bacteria are killed to obtain the target Bacillus spores, and then diluted with normal saline to 10 -3 、10 -4 、10 -5 , take 1mL bacterial liquid and spread it on the screening plate (2% glucose, 0.2% CaCl 2 , 0.5%NH 4 NO 3 , 0.05% KCl, 0.05% MgSO 4 .7H 2 O, 0.001% FeSO 4 .7H 2 O, 0.001% MnSO 4 .4H 2O, 0.3% calcium phytate, 2% agar, pH 7.0), use its hydrolyzed calcium phytate to produce transparent circles to screen the target strains, pick the strains with larger transparent circles for streak separation, and obtain their single colonies, Complete the first round of screening. The results of plate screening are shown in the figure figure 1 shown.

[0025] Considering that some bacteria can...

Embodiment 2

[0027] Example 2. Strain identification

[0028] The screened strains were identified by the combination of traditional strain identification and molecular biology identification. Referring to "Bergey's Bacteria Identification Manual" (8th edition), carry out Gram staining and spore staining on ZJ0902 (staining results are as follows: Figure 2A , 2B shown), observed its morphological structure and location of spores, combined with some special physiological responses of Bacillus, designed the corresponding culture medium and a series of biochemical reaction tubes. Based on the physiological and biochemical reactions of the bacterium, it was initially positioned as a Brevibacterium in the genus Bacillus. The physiological and biochemical identification results of the strains are shown in the table below:

[0029] Identification project result Identification project result a ＋ Catalase ＋ Whether to produce spores Square columnar, mesogenic, ...

Embodiment 3

[0035] Example 3. Study on Enzyme Production Performance of ZJ0702 Strain and Optimization of Fermentation Conditions

[0036] The fermentation medium of the ZJ0702 strain was further optimized to improve its ability to produce enzymes. The present invention starts from the common Bacillus enzyme-producing culture medium, selects easy-to-obtain and cheap raw materials, gradually replaces the carbon source, nitrogen source and inorganic salt in the culture medium, and then cultivates the temperature, inoculum size, pH and loading Optimize the culture conditions such as liquid volume, and screen out the most suitable medium components and the best culture conditions to increase the production of phytase. Use part of the factor analysis experiment to scientifically analyze the influence of each factor on enzyme production, determine the main influencing factors, and then determine the experimental center point through the steepest climbing experiment, and finally conduct resea...

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Abstract

The invention discloses a high-yield heat resistant type neutral phytase bacterial strain. The classification naming of the bacterial strain is Bacillusnealsonii, the preservation number is China General Microbiological Center Culture Collection Center (CGMCC) No.5396, the preservation date is October 28th, 2011, and the preservation unit is CGMCC. The phytoenzyme produced by the bacterial strain has better high-temperature heat resistance, 41 percent of activity is also reserved after 30min under warm bath at 80 DEG C, under the condition of 90 DEG C, 73 percent of activity is also reserved after 10min, and 21 percent of activity is reserved after 30min. The invention also provides a bacterial strain culture medium and a method for producing heat resistant type neutral phytase by the bacterial strain.

Description

technical field [0001] The invention belongs to the field of microorganisms, and in particular relates to a high-yield heat-resistant neutral phytase strain, a fermentation medium thereof and a method for producing heat-resistant neutral phytase. Background technique [0002] Phytic acid is a kind of inositol hexaphosphate substance, the chemical name is cyclohexanol-1, 2, 3, 4, 5, 6-dihydrogen hexaphosphate, it is a light yellow viscous liquid, widely present in Cereals, legumes and oil crops are the main storage form of phosphorus. Phytic acid has a strong chelating ability, exhibits a strong anti-nutritional effect, and can interact with various metal ions such as Zn 2+ , Ca 2+ 、Cu 2+ , Fe 2+ , Mg 2+ 、Co 2+ Phytic acid can chelate with proteins, amino acids, vitamins, etc. to reduce their solubility and affect animals' digestion and absorption of these nutrients; Acid can also combine with pepsin, amylase, lipase, trypsin, etc. in the digestive tract of animals to ...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N9/16C12R1/07
Inventor 于平
Owner ZHEJIANG GONGSHANG UNIVERSITY
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