CN1212386C

CN1212386C - Process for preparing pectinase by solid fermentation of coom aspergillus mutant K58

Info

Publication number: CN1212386C
Application number: CN 99103291
Authority: CN
Inventors: 刘海森
Original assignee: Individual
Current assignee: Individual
Priority date: 1999-04-01
Filing date: 1999-04-01
Publication date: 2005-07-27
Anticipated expiration: 2019-04-01
Also published as: CN1273273A

Abstract

The present invention relates to a method for producing pectase by the solid fermentation of carbon soot aspergillus mutants K58, which belongs to the technical field of biotechnology. The method comprises the following steps: using wheat bran and bagasse powder as main raw materials to carry out the solid fermentation of the carbon soot aspergillus mutants K58; carrying out leaching by warmed water at 35 to 38 DEG C; separating, removing dregs and clarifying; carrying out concentration by a hollow fibre ultrafilter; detecting pectase vitality to reach operation requirements; finally, subpackaging the pectase to obtain a finished product. K58 strains have the advantages of unique morphological structure, culture characteristics and formula of a culture medium, high enzyme vitality and good use effect. The product of the K58 strains is acidic pectase which is suitable for degumming and clarification in the process of processing various kinds of fruit juice and fruit wine.

Description

Process for preparing pectinase by solid fermentation of coom aspergillus mutant K 58

Process for preparing pectinase by solid fermentation of coom aspergillus mutant K 58 belongs to biological technical field.

Microbial fermentation processes is produced polygalacturonase, be to use the edible bacterial strain of safety non-toxic, the substratum with suitable formulated carries out solid fermentation or liquid fermenting, through technologies such as extracting, separate, be refining, concentrated, make finished product zymin again with high fructose enzymic activity.The purpose of this invention is exactly to produce the pectase preparation of the cheapness that meets the food service requirements with microbial fermentation processes.Particular content is as follows:

One, uses the mutant strain K58 of bacterial strain as carbon black aspergillus [Aspergillus Carbonarius (Bainier) Thom] As3.396.

The morphological structure of this bacterial strain with conidiophore repeatedly branch to become dendroid be its feature (accompanying drawing).Cultural characteristic is to become the point-like growth on Cha Shi flat board or slant medium, forms needle point greatly to the big particle of the Chinese sorghum grain of rice on the wheat bran substratum, does not spread the growth shape and be not a large amount of expansions of former starting strain mycelium, but still keeps stronger product spore ability.This morphological specificity of this bacterial strain and cultural characteristic are that starting strain and other aspergillus niger are not available.

Preservation that the K58 bacterial strain (is called for short CGMCC) on April 1st, 1999 at China Committee for Culture Collection of Microorganisms common micro-organisms center, its deposit number is: CGMCC NO 0389.

Two, production of hybrid seeds substratum and culture condition thereof

1, the slant strains substratum is Cha Shi agar or the Cha Shi agar that adds 5% beet ground-slag.Inoculating back 30 ℃ of incubators cultivated 5-7 days.

2, producing bacterium culture medium consists of:

Wheat bran 90g, beet ground-slag 10g, ammonium sulfate 2g, sal epsom 0.07g, dipotassium hydrogen phosphate 0.1g, water 100ml.PH4.0 (transferring) with sulfuric acid, an amount of packing triangular flask or big Cans, 1kg pressure sterilization 30 minutes.Cultivate 4-5 days spore maturations for 30 ℃ after the inoculation slant strains.

Three, fermention medium and culture condition thereof:

Substratum consists of: wheat bran 55, beet ground-slag 35, apple ground-slag 10, ammonium sulfate 7-11, sal epsom 0.07, dipotassium hydrogen phosphate 0.1, water 100, pH4.0 (transferring with sulfuric acid).1kg pressure sterilization 1 hour is pressed the above microbial strain culture of dry-matter 0.1% amount inoculation.30 ℃ of thick-layers ventilate and cultivate or posture is cultured to spore maturation (about 62-72 hour).

Four, the fermenting-ripening song was with 35-38 ℃ of warm water lixiviate of 5 times of amounts 2 hours, centrifugation is removed slag, the plate-and-frame filter press filtering is clarified or used to clear liquid naturally, concentrate about 50 times with hollow fiber membrane ultrafiltration device, detect to pectinase activity and reach service requirements (5-10 ten thousand units, the reducing sugar that the hydrolysis of Somgyi colorimetric method for determining produces, 1 hour produces the 1mg reducing sugar is 1 enzyme activity unit), be packed as finished product.

Five, the relative merits of this production technique

1, this bacterial classification and explained hereafter polygalacturonase thereof produce the product enzyme activity that enzyme activity is higher than domestic known bacterial classification, and fermentation (100kg) test-results in batches repeatedly reaches 1200-1800u/ml with the pectinase activity of 5 times of water gaging vat liquors of former fermention medium dry-matter.Domestic representational polygalacturonase research data such as Cui Fumian etc.: the seed selection of polygalacturonase CP-85211 bacterial strain and the research of liquid fermentation condition thereof (microorganism journal 27 (1) 37-44,1987) and Liu Zirong etc.: the research of aspergillus niger polygalacturonase (food and fermentation industries (6) 16-22,1987), it produces the product enzyme activity that enzyme activity all is lower than the present invention.It is reported that the vigor of Tianjin zymin factory solid fermentation production polygalacturonase also is lower than the present invention's product enzyme activity (do not see and publish).

2, the polygalacturonase of this explained hereafter is an acid pectase, the suitableeest action pH 3.5-4.5, and optimum temperature is 50-52 ℃, is applicable to come unstuck, clarify use in the processing of various fruit juice, fruit wine.The bulge test sample meets China existing polygalacturonase standard QB1502-92 regulation through Gansu Province's food quality supervision test house and the detection of Shaanxi Province commodity inspection and testing bureau.

3, this process using solid fermentation is cultivated, each growth phase zoon of thalline, and made pectase preparation practical effect is good.Its bulge test sample is economized six tame concentrated Succus Mali pumilae factories and is made (examination) usefulness in producing in Shaanxi etc., effect is fine, fully can import substitutes.

4, this process solids fermentation, technology is simple, invests lessly, be convenient to suitability for industrialized production, and raw materials for production is inexpensive, and product cost is low, is easy to obtain economic benefit.

5, the shortcoming of this technology is that fermentation stage is difficult for accomplishing the absolutesterility cultivation, therefore uses this technology must note the sterile culture of fermentation stage more.

Description of drawings: branched conidiophore of K58 bacterial strain and spore head (* 320)

Claims

1, a kind of bacterial strain of high yield polygalacturonase: the mutant strain K58 of carbon black aspergillus [Aspergillus Carbonarius (bainier) thom] As3.396, the formation structure of this bacterial strain with conidiophore repeatedly branch to become dendroid be its feature, cultural characteristic is to become the point-like growth on Cha Shi flat board or slant medium, on the wheat bran substratum, form needle point greatly to the big particle of the Chinese sorghum grain of rice, do not spread the growth shape and be not a large amount of expansions of former starting strain mycelium, but still keep stronger product spore ability, this bacterial strain is by China Committee for Culture Collection of Microorganisms common micro-organisms center (being called for short CGMCC) preservation, preservation date is on April 1st, 1999, and deposit number is CGMCC NO 0389.

2, the described K58 bacterial strain of claim 1 solid fermentation is produced the method for polygalacturonase, comprise solid fermentation, the warm water lixiviate, separate the clarification of removing slag, hollow fiber membrane ultrafiltration device is concentrated into the finished product polygalacturonase, and it is characterized by in the solid fermentation with wheat bran and beet ground-slag is that the uniqueness of main raw material is produced bacterial classification production of hybrid seeds substratum and fermentative medium formula:

Producing bacterial classification production of hybrid seeds substratum is: wheat bran 90g, beet ground-slag 10g, ammonium sulfate 2g, sal epsom 0.07g, dipotassium hydrogen phosphate 0.1g, water 100ml, pH4.0 (transferring with sulfuric acid)

Fermention medium is: wheat bran 55, beet ground-slag 35, apple ground-slag 10, ammonium sulfate 7-11, sal epsom 0.07, dipotassium hydrogen phosphate 0.1, water 100, pH4.0 (transferring with sulfuric acid).