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A kind of thermophilic bacillus producing maltotetraose amylase and application thereof

A technology of thermophilic bacillus and maltotetraose, applied in the field of microorganisms, to achieve the effects of wide reaction temperature range, extended shelf life, and improved overall texture parameters

Active Publication Date: 2020-06-09
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, there are few researches on the method of preparing maltotetraose in my country.

Method used

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  • A kind of thermophilic bacillus producing maltotetraose amylase and application thereof
  • A kind of thermophilic bacillus producing maltotetraose amylase and application thereof
  • A kind of thermophilic bacillus producing maltotetraose amylase and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] The screening of embodiment 1 bacterial strain

[0043] The soil will be collected from different places in different regions (garbage dumps, starch factories, vegetable markets, food factories, grain processing factories, hotels, etc.), take 2g of soil samples and mix them with 9mL of sterile water, add glass beads and shake well After standing still for a while, take 5mL of supernatant and add it to LB medium, shake and culture at 25°C-37°C, 200r / min for 2-3d. Take 10mL of the culture solution and add it to another fresh LB medium to continue the shaking culture. After repeating this operation twice, spread it on the LB solid plate and culture it upside down at 25°C-37°C for 2-3d. A single colony can be observed and screened A strain that grows well.

[0044] A single colony was selected and inoculated into a high-throughput shallow-well plate filled with LB seed medium, and cultured overnight at 37°C with shaking at 200r / min. Then, according to the inoculum amount ...

Embodiment 2

[0051] The preparation of embodiment 2 maltotetraose

[0052] With 50mmol / L, pH 6.0, 7.0, 7.5, 8.0, 9.0 Na 2 HPO 4 -The final concentration of citric acid buffer preparation is 15% (W / V) maltodextrin, the addition amount of maltotetraose amylase fermentation broth is 30U / g maltodextrin, and the common maltodextrin of 5U / g is added Lulanase, 45°C constant temperature water bath shaker, 150rpm, reacted for 12h. The enzyme conversion product was detected by HPLC.

[0053] The results showed that the enzyme-catalyzed maltotetraose changed significantly with the increase of pH. When the pH was 7.5, the yield of maltotetraose reached 62.9%. Excessive pH will reduce the yield of maltotetraose (see figure 1 ).

Embodiment 3

[0054] The preparation of embodiment 3 maltotetraose

[0055] Use 50mmol / L, pH 7.5 Na 2 HPO 4 -The final concentration of citric acid buffer preparation is 15% (W / V) maltodextrin, the addition amount of maltotetraose amylase fermentation broth is 30U / g maltodextrin, and the common maltodextrin of 5U / g is added For Lulanase, the temperature was 40, 45, 50, 55, 60°C, constant temperature water bath shaker, 150rpm, and reacted for 12h. The enzyme conversion product was detected by HPLC.

[0056] The results showed that the temperature had a great influence on the yield of maltotetraose. When the temperature was 45℃, the yield of maltotetraose reached 63.2%. Continue to increase temperature after this, the yield of maltotetraose begins to decline (see figure 2 ). When the reaction temperature is about 45°C, the higher temperature can prevent the reaction system containing a large amount of sugar from being affected by bacterial contamination, improve the enzyme conversion ef...

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Abstract

The invention discloses a thermophilic bacillus and an application thereof, belonging to the technical field of fermentation engineering. The invention obtains a strain of thermophilic bacillus expressing maltotetraose amylase, uses maltodextrin as a substrate, and uses strain fermentation liquid as a catalyst to synthesize maltotetraose. In the present invention, the maltodextrin with a final concentration of 20g / 100mL is used as the substrate, the amount of maltotetraose amylase fermentation liquid added is 40U / g maltodextrin, and the enzyme reaction is carried out at pH 7.5 and temperature 45°C for 18 hours. The conversion rate of maltotetraose can reach 65.7%.

Description

technical field [0001] The invention relates to a thermophilic bacillus producing maltotetraose amylase and an application thereof, belonging to the technical field of microorganisms. Background technique [0002] Maltotetraose is a straight-chain maltooligosaccharide composed of four α-D glucose groups connected by α-1,4 glycosidic bonds, and is a functional food. Maltotetraose has low sweetness and high viscosity, 20% of the sweetness of sucrose, and 2-5 times the viscosity of sucrose. It can replace foods with high sweetness and high calories; it has good moisture retention, strong thickening effect, acid resistance, It is heat-resistant and easy to form a glossy film, which can be used in the papermaking field; maltotetraose syrup products are easy to digest and absorb, and have low osmotic pressure; it can promote the body's ability to absorb Ca 2+ Absorption; it can also inhibit the effect of Clostridium aerogenes, which is an intestinal endocrine toxin, which helps t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N9/26C12P19/14C12P19/00A21D8/04A61P1/00C12R1/07
CPCA21D8/042A61P1/00C12N1/20C12N9/2411C12P19/00C12P19/14C12Y302/0106C12N1/205C12R2001/07
Inventor 吴敬宿玲恰杨亚楠
Owner JIANGNAN UNIV
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