Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Mutant strain of aspergillus sojae with enhanced protease activity and preparation method of natural taste enhanced using the same

A technology of protease activity and Aspergillus sojae, which is applied in the field of Aspergillus sojae mutant strains, can solve the problems of low activity level and so on

Active Publication Date: 2012-07-11
CJ CHEILJEDANG CORP
View PDF7 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, most of them have low activity grades

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Mutant strain of aspergillus sojae with enhanced protease activity and preparation method of natural taste enhanced using the same
  • Mutant strain of aspergillus sojae with enhanced protease activity and preparation method of natural taste enhanced using the same
  • Mutant strain of aspergillus sojae with enhanced protease activity and preparation method of natural taste enhanced using the same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1: Isolation and selection of bacterial strains

[0028] Serially dilute 5 g of each sample, such as fermented soybean pieces, soybean paste (traditional bean paste), soy sauce, etc. collected from all over the country; add 0.1% Tween 80; then apply the diluted sample to On minimal agar plates (Charlotte agar) containing 0.1% Triton X-100. It was cultured at 30° C. for 3 to 4 days, and then a strain having a large clear zone around the colony was isolated.

[0029] The isolated strains were inoculated again in a minimal liquid medium (Chasse Broth; 0.3% NaNO 3 , 0.1%K 2 HPO 4 , 0.05% MgSO 4 .7H 2 O, 0.05% KCl, 0.001% FeSO 4 .7H 2 O, 3% sucrose), and then cultivated at 30°C for 3 days to 4 days. Cells were removed by centrifugation, the supernatant was collected, and then the proteolytic capacity was determined and compared by using casein as a substrate.

[0030] Among the strains, a strain with excellent proteolytic ability was selected, inoculated on...

Embodiment 2

[0032] Embodiment 2: the improvement of aspergillus sojae bacterial strain

[0033] (1) NTG mutagenesis

[0034] The above-mentioned Aspergillus sojae CJCC_011057P (KCCM 11043P) was cultured on a PDA slant medium to obtain a spore suspension. A 3 mg / ml solution of N-methyl-N'-nitroso-N-nitrosoguanidine (NTG) was mixed with an equal amount of the spore suspension, followed by incubation at 30°C for 1 hour.

[0035] The spore suspension was spread on PDA medium to select surviving colonies, and then the selected surviving colonies were cultured on PDA slant medium, and cultured at 30° C. for 4 to 7 days. At least 1,000 mutants were selected, and then cultured at 30°C in a liquid medium (3% defatted soybean, 0.5% KH 2 PO 4 , 1.5% yeast extract, 1.5% glucose, 0.05% magnesium sulfate) for 48 hours to measure protease activity. According to the results of protease activity determination, the strains with high activity were selected, and then the mutations of the strains were con...

Embodiment 3

[0038] Embodiment 3: measure the protease activity of the mutant strain that washes and selects

[0039] Protease activity was determined using casein as a substrate according to the method of Anson-Hagihara (B. Hagihara et al., 1958, J. Biochem. 45 (1958), 185-94).

[0040] Mix 400 μl of 0.75% casein solution and 100 μl of 0.24M Na 2 HPO 4 The solution (pH 7.5) was preincubated at 37°C for 5 minutes, and 100 µl of the culture solution of each selected mutant strain was added; then incubated at 37°C for 10 minutes. 600 μl of a mixture solution mixed with equal amounts of (1) 0.1M TCA (2) 0.22M sodium acetate and (3) 0.33M acetic acid was added to stop the reaction. 200 μl of the supernatant of the reaction solution was added to 500 μl of 0.55 M sodium carbonate solution, and then 100 to 500 μl of a commercially available phenol reagent diluted twice was added, followed by stirring. Thereafter, it was incubated at 30° C. for 30 minutes, and the absorbance was measured at 6...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The present invention relates to an Aspergillus non-inherited genetic variant having enhanced protease activity, and to a production method for a natural flavour enhancer employing the same, and more specifically relates to: an Aspergillus non-inherited genetic variant having enhanced protease activity which is obtained by selecting a strain having a high level of protease activity inducing genetic variation by treating the same with NTG (N-methyl-N'-nitroso-N-nitrosoguanidine) and exposing the same to radiation; and to a production method for a natural flavour enhancer employing a hydrolysed protein obtained by culturing the Aspergillus non-inherited genetic variant, adding the resulting culture product to a protein source and allowing hydrolysis to proceed.

Description

technical field [0001] The invention relates to a mutant strain of Aspergillus sojae with enhanced protease activity and a method for preparing a natural taste enhancer using the mutant strain. Background technique [0002] In the preparation of soy sauce, soybean paste (traditional fermented bean paste) and the like, Aspergillus oryzae and Aspergillus sojae, which are Aspergillus genus, are used to produce enzymes such as protease, amylase, glutaminase and the like. Proteases degrade proteins into peptides and amino acids, and when degrading proteins, glutaminase plays an important role in seasoning soy sauce, soybean paste, etc. by converting free glutamine into tasty glutamic acid. [0003] Therefore, many studies have been conducted to improve the enzymatic activity of protease or glutaminase, which play an important role in seasoning. The method of microbial mutation has following several: by ultraviolet irradiation, mutagen treatment and the radiation (H.Sekine etc., ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/01C12N1/20A23L27/20A23L27/21
CPCC12P21/06C12R1/66C12N1/145C12R2001/66A23L27/20C12N1/20
Inventor 池玄赵成捘
Owner CJ CHEILJEDANG CORP
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com