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A kind of production method of rhamnosidase

A technology of rhamnosidase and production method, which is applied in the field of rhamnosidase production, can solve the problems of high production cost, low yield, low fermentation activity, etc., and achieve the effects of high production efficiency, easy storage and simple process

Active Publication Date: 2019-01-18
宁夏乙征生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Its industrial production is produced by microbial fermentation, but this method has restrictive factors such as low fermentation activity, low yield, and high production cost. Therefore, in order to overcome the above problems, technicians use various genetic methods to improve the production performance of microorganisms. Much work has been done, but little has been done to improve fermentation performance through improvements in culture methods

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] A production method of rhamnosidase, comprising the following steps:

[0038] (1) Take Chaetomium spp. to inoculate in a primary seed medium with an inoculum amount of 5%, cultivate at 38°C for 30 hours, control dissolved oxygen above 60%, and cultivate to obtain a primary seed liquid.

[0039] Among them, the ingredients of the primary seed medium are: hesperidin 60g / L, yeast extract 15g / L, sodium chloride 3g / L, ammonium dihydrogen phosphate 3g / L, magnesium sulfate 1g / L, zinc chloride 0.2g / L, calcium propionate 20g / L.

[0040] (2) Inoculate the primary seed liquid into the secondary seed medium with an inoculation amount of 5, cultivate at 38° C. for 30 h, control the dissolved oxygen above 60%, and cultivate to obtain the secondary seed liquid.

[0041] Among them, the ingredients of the secondary seed medium are: hesperidin 60g / L, yeast extract 15g / L, sodium chloride 3g / L, ammonium dihydrogen phosphate 3g / L, magnesium sulfate 1g / L, zinc chloride 0.2g / L, calcium propi...

Embodiment 2

[0047] A production method of rhamnosidase, comprising the following steps:

[0048] (1) Get the Chaetomium spp. inoculation amount of 20% and inoculate it in the primary seed medium, cultivate it at 25° C. for 10 h, control the dissolved oxygen above 60%, and cultivate to obtain the primary seed liquid.

[0049] Among them, the ingredients of the primary seed medium are: hesperidin 50g / L, yeast extract 10g / L, sodium chloride 3 / L, ammonium dihydrogen phosphate 2g / L, magnesium sulfate 0.5g / L, chloride Zinc 0.2g / L, calcium propionate 20g / L.

[0050] (2) Inoculate the primary seed liquid into the secondary seed medium with an inoculum amount of 5%, cultivate at 25° C. for 30 h, control the dissolved oxygen above 60%, and cultivate to obtain the secondary seed liquid.

[0051] Among them, the ingredients of the secondary seed medium are: hesperidin 50g / L, yeast extract 10g / L, sodium chloride 3g / L, ammonium dihydrogen phosphate 2g / L, magnesium sulfate 0.5g / L, chloride Zinc 0.2g / L...

Embodiment 3

[0057] A production method of rhamnosidase, comprising the following steps:

[0058] (1) Take Chaetomium spp. to inoculate in primary seed culture medium with 10% inoculation amount, cultivate at 30° C. for 20 h, control dissolved oxygen above 60%, and cultivate to obtain primary seed liquid.

[0059] Among them, the ingredients of the primary seed medium are: hesperidin 60g / L, yeast extract 15g / L, sodium chloride 3g / L, ammonium dihydrogen phosphate 2g / L, magnesium sulfate 1g / L, zinc chloride 0.3g / L, calcium propionate 25g / L.

[0060] (2) Inoculate the primary seed liquid in the secondary seed medium with an inoculum amount of 5%, cultivate it at 35° C. for 20 h, control the dissolved oxygen above 60%, and cultivate to obtain the secondary seed liquid.

[0061] Among them, the ingredients of the secondary seed medium are: hesperidin 55g / L, yeast extract 13g / L, sodium chloride 4g / L, ammonium dihydrogen phosphate 2g / L, magnesium sulfate 1g / L, zinc chloride 0.4g / L, calcium prop...

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Abstract

The invention provides a production method of rhamnosidase. The production method comprises steps as follows: (1) a first-level seed culture medium is inoculated with Chaetomium gracile strains, and a first-level seed solution is obtained through culture; (2) a second-level seed culture medium is inoculated with the first-level seed solution, and a second-level seed solution is obtained through culture; (3) a fermentation culture medium is inoculated with the second-level seed solution, a mixed culture medium is obtained, then saccharose is added, rhamnose is added in a fed-batch manner, and fermentation broth with the enzyme activity higher than 500 U / mL is obtained; (4) the fermentation broth is filtered, a filtrate is subjected to ultrafiltration and concentration, and a rhamnosidase solution with the enzyme activity higher than 1,250 U / mL is obtained; (5) the rhamnosidase solution is subjected to spray-drying with starch serving as a supporter, and the solid rhamnosidase with the enzyme activity higher than 3,500 U / mL is obtained. The production method of the rhamnosidase adopts the simple technology and is easy to operate and high in production efficiency, and the produced rhamnosidase has the high activity and the good stability and is easy to store.

Description

technical field [0001] The invention relates to the field of rhamnosidase, in particular to a production method of rhamnosidase. Background technique [0002] Rhamnosidase is a hydrolase. Rhamnosidase has a wide range of sources and is widely distributed in bacteria and fungi in nature. The domestic research on rhamnosidase mainly focuses on the rhamnosidase produced by metabolism. There are relatively many foreign studies, mainly including Aspergillus niger, Aspergillus basilica, Aspergillus aculeatus, Penicillium, Bacteroides and Thermomicrobiale. Rhamnosidase has many potential applications in practical processes, such as industrially used to remove the bitterness of naringin in citrus juice, and to hydrolyze hesperidin into rhamnose and hesperetin glycosides. Among them, hesperetin glycoside is an important precursor for industrial production of sweeteners. In addition, rhamnosidase can act on terpene-based glycosides, and has important applications in improving the a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/26C12R1/645
CPCC12N9/2402C12Y302/0104C12Y302/01043
Inventor 陈龙王双平
Owner 宁夏乙征生物工程有限公司
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