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Method for producing umami peptide by staging fed-batch fermentation

A technology of fed-feed fermentation and umami-flavored peptides is applied in the field of segmented feed-feed fermentation to produce umami-flavored peptides, which can solve problems such as difficulties in industrial production of umami-flavored peptides, and achieve the effects of avoiding substrate inhibition and increasing yield.

Active Publication Date: 2019-05-03
HENGYANG NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Aiming at the above-mentioned prior art, the present invention provides a method for producing umami peptides by staged fed-batch fermentation to solve the difficult problem of industrial production of umami peptides

Method used

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  • Method for producing umami peptide by staging fed-batch fermentation
  • Method for producing umami peptide by staging fed-batch fermentation
  • Method for producing umami peptide by staging fed-batch fermentation

Examples

Experimental program
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Effect test

Embodiment 1

[0020] Example 1: Construction of highly efficient self-induced production strains of umami peptides

[0021] Using the umami peptide recombinant plasmid vector constructed in the patent CN108148116A as a PCR template, the homology arm design of the self-inducible promoter srfA was carried out by using the whole plasmid PCR, so as to construct a recombinant plasmid for highly self-inducible expression of umami peptide. The gene sequence of the self-inducible promoter srfA is shown in SEQ ID NO.1, and the design results of the homology arm sequence of the whole plasmid PCR are shown in Table 1. The umami peptide self-inducible expression vector was obtained by using the whole plasmid PCR amplification technology to obtain Vector plasmid pMA09srfA-10SSRDEQSR with an autoinducible expression promoter.

[0022] Table 1 Homology arm sequence of whole plasmid PCR

[0023]

[0024] Prepare B. subtilis WB800 competent cells by starvation chemical method, add 10 μl of freshly const...

Embodiment 2

[0026] Example 2: Production of umami peptides by multi-stage μ value regulation index fed-batch fermentation

[0027] 1. Culture medium used for production

[0028] LB solid medium (g / L): tryptone 10, yeast extract 5, sodium chloride 10, agar powder 15.

[0029] LB liquid medium (g / L): tryptone 10, yeast extract 5, sodium chloride 10.

[0030] Basic fermentation medium (g / L): diammonium hydrogen citrate 7.5, sodium sulfate 2.0, ammonium sulfate 20.0, ammonium chloride 3.5, dipotassium hydrogen phosphate 14.6, sodium dihydrogen phosphate monohydrate 4.0, magnesium sulfate heptahydrate 1.0, Trace metal ion solution 3.0mL, glucose 20.0 (sterilized separately).

[0031] Trace metal ion solution (g / L): calcium chloride 0.5, zinc sulfate heptahydrate 0.18, manganese sulfate monohydrate 0.1, disodium edetate 10.05, ferric chloride 8.35, copper sulfate pentahydrate 0.16, chlorine hexahydrate Cobalt 0.18.

[0032] Feed medium (g / L): glucose 800.

[0033] 2. Expansion of seed cult...

Embodiment 2

[0040] Table 2 Example 2 multi-stage μ value control strategy

[0041]

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Abstract

The invention discloses a method for producin an umami peptide by staging fed-batch fermentation. The method comprises the following steps: activating a umami peptide-producing strain; inoculating theactivated umami peptide-producing strain into an LB liquid culture medium, and performing enlarge cultivation to obtain a seed culture solution; and inoculating the seed culture solution into a basicfermentation culture medium, performing culturing, starting exponential fed-batch control when the carbon source concentration in the basic culture medium is 5g / L or less, performing exponential fed-batch fermentation at a speed in a formula as shown in the specification, and performing autoinduction to produce the umami peptide. By adopting the method, the requirement of a strain for the carbonsource can be sufficiently met, substrate inhibition caused by carbon source accumulation can be effectively avoided, the yield of a target product can be remarkably increased, and theoretical foundation is laid for large-scale production of the umami peptide and protein.

Description

technical field [0001] The invention belongs to the technical field of biological fermentation, and in particular relates to a method for producing umami peptides by subsection fed-batch fermentation. Background technique [0002] Umami is one of the five basic tastes, which can make people have a pleasant taste experience. Umami peptide is a new type of peptide umami agent after sodium glutamate, guanylic acid and inosinic acid. It not only has a delicious taste, but also provides amino acid nutrients. Among a large number of umami taste peptides, the umami taste peptide with the amino acid sequence Ser-Ser-Arg-Asp-Glu-Gln-Ser-Arg (SSRDEQSR) has significant umami characteristics, which can not only be used alone for food seasoning, but also As a precursor, it participates in the Maillard reaction with sugars to produce a mellow umami taste and a special aroma, which is used to increase the freshness and aroma of food. Therefore, the umami peptide has a huge market potentia...

Claims

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Application Information

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IPC IPC(8): C12P21/02C12R1/125
Inventor 孙伟峰伍圆明王莉伍伦杰陈晓华刘义车振明
Owner HENGYANG NORMAL UNIV
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