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Bacillus circulans and application for same in preparation for ferulic acid decarboxylase

A technology of Bacillus circulans and ferulic acid decarboxylase, which is applied in the field of bioengineering, can solve the problems of low ferulic acid decarboxylase activity and difficult industrial production, etc., and achieves fast growth and reproduction speed, short production cycle and stable performance. Effect

Inactive Publication Date: 2015-05-13
QILU UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

According to our research, most of the ferulic acid decarboxylase-producing bacteria in Daqu are a kind of spore-forming Bacillus, but the activity of ferulic acid decarboxylase produced by these microorganisms is low, so it is difficult to carry out industrial production
[0006] At present, due to the relatively low activity of ferulic acid decarboxylase produced by wild-type microorganisms, ferulic acid decarboxylase genetically engineered bacteria are also in the research stage, and the production of ferulic acid decarboxylase has not yet formed an industrial scale, so it is obtained through mutation breeding technology. Highly active ferulic acid decarboxylase-producing strains and the realization of large-scale production of ferulic acid decarboxylase are very necessary

Method used

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  • Bacillus circulans and application for same in preparation for ferulic acid decarboxylase
  • Bacillus circulans and application for same in preparation for ferulic acid decarboxylase

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Bacillus circulans ( Bacillus circulans ) HG12 strain has the typical characteristics of circular Bacillus: the colony is milky white, wrinkled, dull, and the surface is not smooth; the bacteria are rod-shaped, straight or nearly straight, single or in chains, and the cell size is about (0.6- 1.0) μm×(1.8-3.8) μm, spore-forming, cyst-inflated, spore-elliptic, spore-endotrophic; Gram-positive, facultative anaerobic, organic chemoheterotrophic bacteria, utilizing glucose, wood Sugar, mannitol, and arabinose produce acid, V-P reaction is negative, catalase positive, nitrate reduction positive, can hydrolyze casein, gelatin and starch, do not use citrate and propionate, grow at a culture temperature of 55°C ; Liquid medium and solid medium made from one or more combinations of fermented wheat bran, wheat, barley, sorghum to produce a distinct aroma of sauce.

Embodiment 2

[0037] Bacillus circulans ( Bacillus circulans ) Isolation and screening of HG12 strains, the steps are as follows:

[0038] (1) The bacterial strain involved in the present invention is derived from the high-temperature Daqu used to produce Maotai-flavored liquor "Guizhou Anjiu". Weigh 10 g of uniformly crushed high-temperature Daqu powder in 20 mL of sterile saline, shake at 37 °C and 150 r / min for 24 h to obtain a bacterial suspension; put the bacterial suspension into a water bath at 80 °C and heat for 20 min; then take An appropriate amount of the above-mentioned bacterial suspension was aseptically inserted into the broth medium, and cultured at 37°C and 150r / min for 18 hours to obtain a culture solution; after the above-mentioned culture solution was treated at 80°C for 20 minutes, it was cooled and diluted by 10 times. After appropriate dilution, spread on nutrient agar medium plate, and culture at 37°C for 48h. Colonies with better growth and different traits were ...

Embodiment 3

[0048] Bacillus circulans ( Bacillus circulans ) Stability experiment of ferulic acid decarboxylase produced by strain HG12, the steps are as follows:

[0049] Transfer the bred Bacillus circulans HG12 strains to the slant medium in sequence, subculture 5 times in total, and culture at 37°C for 24 hours to prepare test tube slant surfaces with different generations; Take 1 loop of each strain and inoculate it into the liquid seed medium, and culture it for 16 hours at 37°C and 150 r / min to obtain activated liquid seeds; inoculate the prepared liquid seeds at 5% (v / v) Amounts were inoculated in liquid fermentation medium, and cultured for 48 hours at 40°C and 200r / min. Measure the ferulic acid decarboxylase activity in the fermented liquid of bacterial strain of different algebra (the result is as follows figure 2 shown), with the strain H1 as the control, the bred Bacillus circulans HG12 strain has a relatively stable performance of producing ferulic acid decarboxylase.

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Abstract

The invention belongs to the technical field of bioengineering, and particularly discloses a bacillus circulus and its application in preparing ferulic acid decarboxylase. The Bacillus circulans is characterized in that: the Bacillus circulans (Bacillus circulans) HG12 strain is isolated from the high-temperature Daqu used for brewing Maotai-flavored liquor in a Chinese liquor factory, and is obtained through ultraviolet mutagenesis screening, which has It has fast growth and reproduction speed, can produce ferulic acid decarboxylase under the induction of ferulic acid, and has the characteristics of stable performance and high enzyme activity. The invention can realize the continuous and large-scale production of ferulic acid decarboxylase, facilitate its industrial production, and the enzyme production process is simple, the production period is short, and the cost is low.

Description

(1) Technical field [0001] The invention belongs to the technical field of bioengineering, in particular to bacillus circulans and its application in preparing ferulic acid decarboxylase. (2) Background technology [0002] Ferulic acid decarboxyase (FAD) is an enzyme that catalyzes the decarboxylation of ferulic acid to produce 4-vinylguaiacol. 4-Vinylguaiacol is a volatile phenolic substance that can produce special aroma or clove-like aroma. It can improve the sensory quality of food and has a great effect on the natural aroma of food such as wine, soy sauce, cheese, and beverages. Therefore, the use of ferulic acid decarboxylase in the food industry has a good development prospect. [0003] Studies have shown that ferulic acid decarboxylase exists in many fungi, yeast and some bacteria, such as Brevibacterium ( Bacillus pumilus ), Lactobacillus plantarum ( Lactobacillus plantarum ), Bacillus subtilis ( Bacillus subtilis ),Saccharomyces cerevisiae( Saccharomyces c...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N9/88C12R1/09
Inventor 董永胜
Owner QILU UNIV OF TECH
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