Naringinase production method

A technology for naringinase and seeds, applied in the field of naringinase production, can solve the problems of low immobilization efficiency, complicated equipment, limited application, etc., and achieve the improvement of aroma components, production of food additives, high enzyme production level, and improved flavor. Effect

Inactive Publication Date: 2015-03-11
BEIJING TECHNOLOGY AND BUSINESS UNIVERSITY
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  • Description
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Problems solved by technology

[0004]Although the research on naringinase is getting more and more in-depth, only a small number of naringinases have been industrialized, which currently limits the large-scale industrialization of naringinase There are two problems in the application: the first major problem is that my country lacks high-yield enzyme strains suitable for industrial production, which greatly limits the application. The difficulty in the cultivation of high-yield naringinase strains is how to produce them in a large number of mutagenized offspring. So far, there is no better way to efficiently select excellent mutant strains. The progress of strain selection is slow, and it is necessary to develop new and efficient screening methods. Therefore, the cultivation of high-yield strains is an urgent task at present.
The second problem is that there are still many defects in the immobilization technology of naringinase in China. The reagents or carriers used in the immobilization have high cost, low immobilization efficiency, poor stability, and equip

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1 Decadence and identification of Symblysis 11568 Screening

[0049] 1. JNU002 (BACILLUS AMYLOLOLOLIQUEFACINES) 11568 screening separation

[0050] Called 2G to collect soil -like or moldy or / and pomelo skin from all over the country. It oscillates 30min in a 100ml taper bottle equipped with 18ml physiological saline with sterilization treatment. It is best to put a little glass bead in the cone bottleEssenceThen dilute the volume concentration with sterile water to 1 × 10, respectively -3 , 1 × 10 -4 The dilution of the two gradients is painted with a sterilized gun head in the pre-prepared and sterilized selective flat medium with a sterilized gun head and cultivated 48-96h at 30 ° C.From a selective tablet medium to separate the single -bacterial colonies, some of the selected single -bacterial mirror examination judge the purity of the strain (microscope observes the same morphological characteristics).If it is not a colonies, it should be repeatedly separated f...

Embodiment 2

[0070] Example 2 The preparation of liquid fermentation liquid containing grapemine containing grapemine, hypoprosis 11568 liquid fermentation liquid

[0071] (1) Landscape cultivation: Decadence of starchybacteria 11568 is inoculated to the slash medium, and it is cultivated for 48 hours at 40 ° C.The oblique medium used (G / L) is: MGSO 4 0.5, kh 2 PO 4 1.5, CACL 2 0.1, (NH 4 ) 2 So 4 1.5, KCL 0.5, KNO3 1.5, yeast cream 2, orange peel powder 15, agar 20, pH 6.0, 1 × 10 5 PA sterilization 20 min.

[0072] (2) Liquid bottle fermentation: The obtained strain was inoculated at a vaccination of 0.2%(V / V) to the liquid bottle medium, the cultivation temperature was 40.9 ° C, the speed of the bed was 180R / min, and the cultivation time was 48h; the liquid was obtained;Ferry solution.Liquid shake bottle medium (G / L) is: MGSO 4 0.5, kh 2 PO 4 1.5, CACL 2 0.1, (NH 4 ) 2 So 4 1.5, KCL 0.5, KNO 3 1.5, yeast cream 2, orange peel powder 7.5, pH 6.0, 1 × 10 5 PA sterilization 20 min.

[0073] (3)...

Embodiment 3

[0075] Example 3 Preparation of liquid fermentation liquid with liquid fermentation liquid containing grapepelidinase, deercybaidia 11568

[0076] Step 2 steps (1) The trained strain that is cultivated with 0.2%(V / V) is inoculated into a 250ml cone bottle equipped with a liquid medium, and the liquid bottle is fermented.The bed speed is 180 R / min, the fermentation time is 48 h; the liquid fermentation liquid is obtained.Liquid medium formation (G / L): Pomelochide 10, Tenta 胨 10, (NH 4 ) 2 HPO 4 20, NaCL 10, yeast extract 5; pH 7.51, 1 × 10 5 PA sterilization 20 min.

[0077] (3) The composition of the liquid fermentation liquid is determined through the high -efficiency liquid chromatography method (water: methanol = 1: 1), and the test results are figure 1 : The liquid fermentation liquid contains grapefrofidin and grapefruit skin.

[0078] (4) Measure the vitality of the liquid fermentation liquid prepared by the Example 2 of the Example 2 of the Example 2 of the Example 2.

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Abstract

The invention relates to a naringinase production method and belongs to the technical field of microbe fermentation. The naringinase production method comprises the following steps: bacillus amyloliquefaciens 11568 with the preservation No. of CGMCC No. 9928 is subjected to slant culture to obtain slant strains; under the conditions of 40.9 DEG C and 180 rpm, fermental cultivation is performed for 48 h to obtain a liquid fermentation liquor; the liquid fermentation liquor is subject to centrifugal separation and the supernatant is taken; the supernatant is subject to ammonium sulfate fractional precipitation, ion exchange chromatography and SephacrylS-200 gel filtration chromatography to obtain naringinase. The naringinase production method has the advantages of being high in naringinase production level, short in fermentation time, easy for controlling the yield condition, easy in separation, high in purity, simple to operate, mild in condition and the like, and is completely suitable for industrial production. Crushed orange peel screened by 60 meshes serves as a naringinase fermentation substrate, so that the domestic waste is fully utilized and the cost is lowered.

Description

Technical field [0001] The present invention involves a method for producing grapemine, belongs to the field of microbial fermentation technology. technical background [0002] Grapemine enzyme is an important enzyme for the bitter substances in the juice.-D-GLUCOSIDASE, EC3.2.1.21) A glycoside hydrolytic composite enzyme composite, with the activity of the two enzymes of α-L-rodin and β-D-glucosidase.In the bitter pomelosin and hydrolyzed flavonoids, the process is that the grapefrofidin first generates lip sugar and Prinin (also translated as a cherryinin) under the action of α-L-Ricosanase).The bitterness of Pruning is about one-third of the grapefrontin, so the bitter taste is reduced, and then Puluning can be decomposed as a bitter flavonoids (mainly mainly because the β-D-glucosidaseGrapefrontin).The microorganisms of grapehide are mainly concentrated in mold (black mold and green mold, etc.), and there are fewer reports of bacterial grapemine. [0003] my country is a majo...

Claims

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Application Information

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IPC IPC(8): C12N9/42C12N9/24C12R1/07
CPCC12N9/2402C12N9/2445C12Y302/01021C12Y302/0104
Inventor 李秀婷朱运平杨然伍少明
Owner BEIJING TECHNOLOGY AND BUSINESS UNIVERSITY
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