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Pectinase producing bacterial strain and application in preparation of peeled citrous complex enzyme

A technology of decapsulation compound enzyme and pectinase, which is applied in the direction of enzyme, lyase, hydrolase, etc., can solve the problem of high process cost, achieve stable application, solve the effect of high cost of enzyme, and promote substitution

Active Publication Date: 2013-06-26
重庆檬泰生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] Aiming at the deficiencies in the prior art, the object of the present invention is to provide a citrus decapsulation compound enzyme with low cost, high efficiency, stable application, and product safety, which is especially suitable for tangerines, Wenzhou citrus and other broad-peeled tangerines and its production. method, to solve the problem of high cost of citrus enzymatic decapsulation process

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  • Pectinase producing bacterial strain and application in preparation of peeled citrous complex enzyme
  • Pectinase producing bacterial strain and application in preparation of peeled citrous complex enzyme

Examples

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Embodiment 1

[0045] The strains described in the present invention were screened according to methods known in the art, and a strain of Asperillus niger WZW001 with high pectinase-producing activity and complete enzyme system was obtained, CCTCC No: M2012399.

[0046] Aspergillus niger WZW001 was grown and preserved in PDA medium (test tube slant and eggplant bottle slant), and the enzyme production ability of the strain was stable for more than 6 generations on the slant medium.

[0047] The fermentation process of the present invention is from inclined plane to seed culture medium to 5L fermenter. The strain grows in the inclined plane for 72 hours and matures, and the mature spores are dense and dark brown. Add sterile saline to make spore suspension (1~3×10 7 each / mL), inoculate the seed medium according to the inoculation amount of 5% (v / v), the seed medium formula (100mL): orange peel powder 1g, bran 1g, glucose 2g, ammonium sulfate 1g, MgSO 4 0.05g, before sterilization, adjust th...

Embodiment 2

[0055] Select the citrus decapsulation compound enzyme 1. prepared in example 1 for use in this instance, and the citrus that adopts is Zhejiang Wenzhou Tangerine. The enzyme activities of PL, PE, Exo-PG and FPA prepared by citrus decapsulation complex enzyme ① were 170U, 2400U, 0.83×10 5 U and 1.5×10 4 U enzyme solution 5400mL, adjust the pH value to 3.5 with 2mol / L hydrochloric acid. Fresh Wenzhou tangerines were washed, peeled, and split to obtain 3.6 kg of orange petals, which were completely submerged in the enzyme solution for constant temperature enzymolysis. The study found that after 78 minutes, the enzymatic hydrolysis was completed, and the orange complex and capsule coat on the surface of the orange petals were completely removed. Separate the orange petals from the enzyme liquid, wash the orange petals twice with clean water to remove the residual enzyme liquid, recover the enzyme liquid and continue to use it for the next batch of decapsulation treatment, the e...

Embodiment 3

[0057] This example prepares Aspergillus niger WZW001 fermentation broth according to the method of Example 1. After the fermentation broth is filtered through four layers of gauze, it is centrifuged at a high speed of 10000r / min for 7min to obtain 2000mL of enzyme liquid, which is then concentrated by an ultrafiltration membrane with a molecular weight cut-offfigure 1 ) and enzyme activity assays, it was found that although the single enzyme components Exo-PG, PE, and PL still have some heteroproteins, the three have been separated and have no influence on each other, and can be used separately for compound applications.

[0058]The cellulase used in this example was purchased from Novozymes. The cellulase activity (calculated as FPA) is 1000U / g. The ratio of pectin lyase (PL) single enzyme component to cellulase is PL:FPA enzyme activity The complex enzyme of citrus decapsulation coat was obtained by compounding at 1 / 60:1. Use citrus decapsulation compound enzyme ② to prepare...

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Abstract

The invention provides a pectinase producing bacterial strain, namely Asperillus niger WZW001, and an application in preparation of pectinase and peeled citrous complex enzyme. The strain is preserved in the CCTCC (china center for type culture collection) on Oct. 12, 2012, and the preservation No. is CCTCC No: M2012399. The peeled citrous complex enzyme prepared by the Asperillus niger WZW001 has the advantages of low production cost, specificity and efficiency, stable application and the like; during the application, as long as the Asperillus niger WZW001 strain fermentation liquor is used for preparing solid pectinase or is separated and purified to obtain PL, Exo-PG, and PE single enzyme elements, and one or more of the solid pectinase or the single enzyme elements is compounded with marketed cellulose without adding other enzymes, so that the orange capsule can be efficiently removed. According to the pectinase producing bacterial strain and the application, the problem of high cost of enzyme for an enzyme-method capsule removing process at present is solved, the replacement of the process to an acid-alkali chemical method is facilitated, and the energy-saving, emission-reduction, green and efficient production of the orange deep processing industry in the country has practical significance.

Description

(1) Technical field [0001] The invention relates to a pectinase-producing strain—Asperillus niger (Asperillus niger) WZW001, and its application in the preparation of pectinase and citrus decapsulation compound enzyme. (2) Background technology [0002] my country's citrus planting area and output rank first in the world, with an annual output of 2000×10 4 More than t, my country is also the center of production of canned citrus in the world. About 75% of citrus matures in November to December, and the processing period is only 2 to 3 months. The deep processing of citrus can effectively solve the problem of strong seasonality of citrus. Promote the development of my country's citrus deep processing industry and bring huge economic and social benefits. The decapsulation process is a key process in the production of canned citrus and cysts. At present, the launder-type acid-base treatment method is generally used in industrial decapsulation methods at home and abroad. This me...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N9/18C12N9/26C12N9/88C12N9/00C12N9/42C12R1/685
Inventor 魏春汪钊吴鹏飞鄢洪德章银军
Owner 重庆檬泰生物科技有限公司
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