A kind of feeding method of Bacillus licheniformis producing β-mannanase

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 producing β-mannanase by Bacillus licheniformis HDGLJT-01. Background technique [0002] The production of β-mannanase is inseparable from carbon and nitrogen sources. Carbon sources are indispensable as a source of carbon structure for bacterial proliferation and metabolism. At the same time, nitrogen sources, as a necessary raw material for proteases, are also very important. important. Konjac flour has a unique advantage as a cheap and efficient carbon source material to induce β-mannanase production. However, in the laboratory preliminary test, it was found that the enzyme activity of adding 6% (w / v) high-concentration konjac flour to the 5L fermenter at one time did not reach the expected high-concentration and high-enzyme activity effect. 1% (w / v) konjac flour was initially added to the 5L fermenter, and then 2% (w / v) konjac flour was added ...

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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

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