Naringinase production method

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...

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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 equipment used for continuous operation. It is more complex, etc., which requires our country to further develop a simpler and more applicable immobilization method. There have been relevant reports abroad that naringinase has been used in the debittering and processing of citrus and other products, but naringinase has not been used in China. There is no large-scale application in production. The main reason is that there is no commercial naringinase prepared on a large scale in China. In the future, we should strengthen the method of breeding high-yield naringinase strains and prepare low-cost pure naringinase , has important practical significance

However, compared with mold, the activity of naringinase produced by bacteria still has a certain gap; the reported level of enzyme production by bacterial fermentation does not exceed 1000 U units per milliliter of fermentation broth; the optimum temperature for bacterial fermentation to produce naringinase is moderate Strong environment may limit its application in acidic environment

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