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Material supplementing method for bacillus licheniformis production of beta-mannase

A technology of Bacillus licheniformis and mannanase, which is applied in the field of enzyme fermentation feed, which can solve the problems of unfavorable accumulation of β-mannanase, reduction of dissolved oxygen in the medium, and variable viscosity.

Inactive Publication Date: 2016-09-14
HEILONGJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This is due to the special nature of konjac flour, which is easily viscous when exposed to water. If a large amount of konjac flour is added at one time, the dissolved oxygen in the medium will be reduced, and the proliferation of Bacillus licheniformis will be inhibited, which is not conducive to β-mannose Accumulation of glycanase, and it is easy to cause damage to machinery; however, only a small amount of konjac flour is added at one time, and it cannot provide sufficient carbon source substances for bacterial proliferation and metabolism. Therefore, the research on fed-batch fermentation strategy has become the current laboratory research The key link in the production of β-mannanase by Bacillus licheniformis HDGLJT-01

Method used

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  • Material supplementing method for bacillus licheniformis production of beta-mannase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Effect of carbon source on Bacillus licheniformis (B. licheniformis) fermentation to produce β-mannanase and optimization of additional fermentation strategy

[0042] 1.1 Primary screening of carbon source types

[0043] Based on the fermented enzyme production medium, change the type of carbon source, add 1% (w / v) carbon source glucose, konjac flour and flour, respectively, insert 6.7% (v / v) seed culture solution, 37 ° C, speed 300r / min, 100mL / 250mL shake flasks were cultured for 48 hours, and the enzyme activity of β-mannanase was used as the detection index to determine the optimal carbon source for the enzyme production of the strain HDGLJT-01.

[0044] Depend on figure 1 It can be seen that when the initial carbon source medium is konjac flour, glucose, and flour, the highest values ​​of β-mannanase enzyme activity are 696.35±23.47U / mL, 288.13±21.59U / mL, and 65.21±5.78U / mL, respectively. Since both konjac flour and glucose can promote the fermentation ...

Embodiment 2

[0075] Example 2 Effect of Nitrogen Source on Bacillus licheniformis Fermentation to Produce β-Mannanase and Determination of Supplementary Fermentation Strategy

[0076] 2.1 Preliminary screening of nitrogen source types

[0077] Based on the fermented enzyme production medium, change the type of nitrogen source, add 1% (w / v) (NH 4 ) 2 SO 4 , ammonia, NH 4 Cl, NH 4 NO 3 , yeast powder, beef extract, peptone, bran, urea, soybean meal, inserted into 6.7% (v / v) seed culture solution, 37 ° C, 300r / min, 100mL / 250mL liquid volume shake flask culture for 48h, with β- The enzyme activity of mannanase was used as the detection index to determine the optimal nitrogen source for strain HDGLJT-01 to produce enzyme.

[0078] In this experiment, considering the effect of inducing β-mannanase and economic factors, NH 4 NO 3 , ammonia, NH 4 Cl, (NH 4 ) 2 SO 4 , yeast powder, beef extract, peptone, bran, urea, and soybean meal were used as nitrogen source materials for HDGLJT-01...

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Abstract

A material supplementing method for bacillus licheniformis production of beta-mannase comprises the following steps of carrying out shake-flask culture at the temperature of 37 DEG C with a fermentation enzyme production culture medium as a base. The fermentation enzyme production culture medium contains a nitrogen source and a carbon source, the nitrogen source comprises NH4NO3, ammonia water, NH4Cl, (NH4)2SO4, a yeast powder, a beef extract, peptone, bran, urea, soybean meal or a combination thereof, and preferably, the nitrogen source is a combination of two or more components; the carbon source is a konjac powder, glucose, flour or a combination thereof, and preferably, the carbon source is a combination of two or more components; the initial adding amount of the nitrogen source is 0.1-10% (w / v); the initial adding amount of the carbon source is 0.1-10% (w / v); supplementing is performed in 1-40 hours after the reaction is started or when the enzyme activity is reduced or the number of bacteria is reduced or reducing sugar is reduced, and preferably, during the supplementing, the carbon source and the nitrogen source are added simultaneously, or the carbon source and the nitrogen source are added at different time; the supplementing amount of the nitrogen source is 0.1-10% (w / v); and the supplementing amount of the carbon source is 0.1-10% (w / v).

Description

technical field [0001] The invention relates to a feeding method for enzyme fermentation, in particular to a feeding method for beta-mannanase produced by bacillus licheniformis HDGLJT-01. Background technique [0002] The production of β-mannanase is inseparable from carbon source and nitrogen source. Carbon source is indispensable as the source of carbon frame composition necessary for cell proliferation and metabolism. At the same time, nitrogen source is a necessary raw material for proteases. important. Konjac flour has a unique advantage as a cheap and efficient carbon source material for inducing β-mannanase production. But find in laboratory preliminary test: in 5L fermentor tank, one-time adds the high-concentration konjac powder of 6% (w / v), its enzyme activity has not reached the effect of expected high-concentration high enzyme activity. Adding 1% (w / v) konjac flour initially in a 5L fermenter, and then adding 2% (w / v) konjac flour in the late logarithmic growt...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/24C12R1/10
CPCC12N9/2494C12Y302/01078
Inventor 葛菁萍平文祥赵丹杜仁鹏金曼
Owner HEILONGJIANG UNIV