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A kind of preparation method of mannanase and its extraction and refining device

A technology of mannanase and bacteria strains, applied in the field of mannanase, can solve the problems of only 80% of the finished product hygiene index pass rate, only 70% of the product yield, and poor on-site environment, so as to expand product sales The effect of increasing the scope, enhancing product value, and increasing the qualified rate

Active Publication Date: 2021-01-22
HEC PHARM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The traditional post-extraction process uses plate and frame filtration to separate bacteria, which requires the addition of a large amount of flocculant, high labor intensity, poor on-site environment, and long filtration time. The qualified rate of final product hygiene indicators is only 80%, and the product yield is only 70%. %

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  • A kind of preparation method of mannanase and its extraction and refining device

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] A method for extracting and refining mannanase, the steps are:

[0033] 1) The fermented liquid is concentrated by circulating filtration with a ceramic membrane, and the temperature of the circulating feed liquid is controlled below 20°C; the pore size of the ceramic membrane is 100nm, the operating pressure difference is 0.40MPa, and the membrane surface flow rate is 3.5m / s. The filtrate was clear and transparent, with a slag content < 0.05%, and a yield of 88.8%.

[0034] 2) After concentrating for 4 hours, the filtrate filtered by the ceramic membrane is circulated and concentrated by ultrafiltration membrane equipment, and the temperature of the circulating feed liquid is controlled below 20°C; the ultrafiltration membrane is an organic membrane with a molecular weight cut-off of 10,000 Daltons. The concentrated solution obtained by ultrafiltration is still clear and transparent, the volume is reduced to 1 / 5 of the filtrate volume, and the yield is 97.5%.

[0035]...

Embodiment 2

[0037] A method for extracting and refining mannanase, the steps are:

[0038] 1) The fermented liquid is concentrated by circulating filtration with a ceramic membrane, and the temperature of the circulating feed liquid is controlled below 20°C; the pore size of the ceramic membrane is 200nm, the operating pressure difference is 0.40MPa, and the membrane surface flow rate is 2.5m / s. The filtrate is clear and transparent, with a slag content < 0.07%, and a yield of 89.3%.

[0039] 2) After concentrating for 8 hours, the filtrate filtrated by the ceramic membrane is circulated and concentrated by ultrafiltration membrane equipment, and the temperature of the circulating feed liquid is controlled below 20°C; the ultrafiltration membrane is an organic membrane with a molecular weight cut-off of 10,000 Daltons. The concentrated solution obtained by ultrafiltration is still clear and transparent, and the volume is reduced to 1 / 7 of the filtrate volume, and the yield is 97.3%.

[0...

Embodiment 3

[0042] A method for extracting and refining mannanase, the steps are:

[0043] 1) The fermented liquid is concentrated by circulating filtration with a ceramic membrane, and the temperature of the circulating feed liquid is controlled below 20°C; the pore size of the ceramic membrane is 500nm, the operating pressure difference is 0.30MPa, and the membrane surface flow rate is 2.5m / s. The filtrate is clear and transparent, with a slag content < 0.08%, and a yield of 89.7%.

[0044]2) After concentrating for 6 hours, the filtrate filtered by the ceramic membrane is circulated and concentrated by ultrafiltration membrane equipment, and the temperature of the circulating feed liquid is controlled below 20°C; the ultrafiltration membrane is an organic membrane with a molecular weight cut-off of 20,000 Daltons. The concentrated solution obtained by ultrafiltration is still clear and transparent, and the volume is reduced to 1 / 8 of the filtrate volume, and the yield is 97.0%.

[004...

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Abstract

The invention discloses a mannase preparation method and an extraction and refining device of mannase. The mannase preparation method comprises the following steps: firstly, performing liquid fermentation to pichia pastoris engineering strain to obtain a fermented liquor, then extracting and refining, the specific extracting and refining steps are as follows: 1) circularly filtering and concentrating the fermented liquor through a ceramic film, wherein the temperature of the cyclic liquor is controlled to be below 20 DEG C; 2) after concentrating for certain time, taking clear filtrate filtered by the ceramic film, and circularly concentrating through an ultrafilter membrane equipment, wherein the temperature of the cyclic liquor is controlled to be below 20 DEG C; and 3) when the volume of concentrated liquor in the step 2) is reduced to 1 / 5-1 / 10 of the clear filtrate, adding starch and evenly mixing, and finally performing spray granulation, to obtain mannase. Adopting the ceramic film for filtering not only can hold up producing strains, but also realize filtration sterilization, and through coupling with the concentration of the ultrafiltration membrane, the technological time can be shortened, the proliferation risk of microorganism can be reduced, the hygienic index pass percent of the obtained finished product is greatly improved to 99%, and the yield of the product is improved to 83% or above.

Description

technical field [0001] The invention belongs to the technical field of mannanase, and in particular relates to a method for preparing mannanase and an extraction and refining device thereof. Background technique [0002] Mannanase (β-1,4-D-Mannanase, EC3.2.1.78) is a hemicellulolytic enzyme that degrades β-1,4-glucosidic bonds in an endo-cutting manner, and the non-reducing end of the degradation product is Mannan oligosaccharides, whose substrates include β-mannan, galactomannan and glucomannan. β-mannanase can also degrade mannan into mannan oligosaccharides. Mannan oligosaccharides can optimize the intestinal micro-ecological flora, enhance the beneficial health function of the host, and can also identify, adhere to and eliminate pathogenic microorganisms, and regulate the body's Immunity function, improve the production performance of livestock and poultry. Mannan oligosaccharides can competitively combine with some pathogenic bacteria in animals to promote the growth ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/24C12N15/81C12M1/12
CPCC12M47/12C12N9/2494C12N15/815C12Y302/01078
Inventor 熊辉刘代云
Owner HEC PHARM
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