CN109679826B

CN109679826B - Siphon catheter feeding and shaking table cultivation method for mold

Info

Publication number: CN109679826B
Application number: CN201811584783.7A
Authority: CN
Inventors: 刘志强; 沈水兴; 沈昊; 刘政
Original assignee: Shanghai Zhicheng Analytical Instrument Manufacturing Co ltd
Current assignee: Shanghai Zhicheng Analytical Instrument Manufacturing Co ltd
Priority date: 2018-12-24
Filing date: 2018-12-24
Publication date: 2022-04-01
Anticipated expiration: 2038-12-24
Also published as: CN109679826A

Abstract

A siphon guide tube material-supplementing shaking table culture method for mould is characterized by that on the basis of material-supplementing shaking table a functional siphon guide tube solid culture connector is added to make several shaking bottles be series-connected, starting from adding target strain into first culture bottle, and making solid-liquid alternative fermentation by siphon guide tube communication until the last shaking bottle is finished so as to obtain the invented quick multiple culture. The method can be used for domesticating and separating the rejuvenation of high-activity pectinase aspergillus niger and beauveria bassiana strains, wherein the materials, the pipe diameter, the length of the pipe, the vector composition preparation in the pipe and the like used by the functional siphon pipe have key influences on the culture of target strains, so the siphon pipe can be specially designed according to different uses; the invention provides a new technical means for domestication, separation and rejuvenation of microbial strains by using the material supplementing shaking table, and is suitable for being widely applied to the fields of food, medicine, feed, biological products and the like.

Description

Siphon catheter feeding and shaking table cultivation method for mold

Technical Field

The invention relates to the technical method of biological research and the field of shaking table equipment manufacturing, in particular to a siphon guide pipe feeding shaking table cultivation method for mould.

Background

The shaking table is an important tool for researching microorganisms by utilizing triangular shaking flask oscillation fermentation, and has a long history and wide application. The feeding table is used for timely feeding nutrition into the triangular shake flask and adjusting the pH of fermentation liquor in the flask to improve the fermentation environment and prolong the fermentation time and increase fermentation products; the feeding endows the traditional shaking table with new functions.

The domestication, separation and rejuvenation of strains are important technical means for the research of microbial culture, and the three are independent and mutually related. Domestication is the first one, and is based on Darwinian biological evolution theory 'environmental selection and fittest survival', and is divided into three different ecological niche adjustment processes, wherein the first ecological niche adjustment is to restore strains by firstly performing liquid culture in a room (bacterial sludge) under the original nutrition state, and then performing liquid re-culture after solid culture can be identified by naked eyes, so that the fitter strains under a new environment are restored for the second time, and the unsuitable strains are eliminated. The second ecological niche adjustment is to change the culture medium environment according to needs, so that more strains which are not suitable for survival are separated and eliminated; the third niche adjustment is the selection of individual strains by changing the culture environment again and the stimulation of their working capacity for production. Each process is that solid culture and liquid shaking table fermentation are alternately carried out and repeated for many times.

Disclosure of Invention

The invention aims to provide a siphon conduit feeding shaking table culture method for mould aiming at the defects of the prior art, which utilizes a multifunctional siphon conduit and a feeding shaking table, arranges a solid culture medium in the conduit and then connects the shaking table, so that the prior solid-liquid alternative culture operation is integrated on the shaking table for one time, and the siphon conduit is selected and designed according to the requirement, thereby being a great technical improvement on the application of the feeding shaking table.

The invention is realized by the following scheme:

a siphon guide tube material-supplementing shaking table culture method for mould is based on a material-supplementing shaking table, wherein the material-supplementing shaking table comprises a plurality of shaking bottles, culture solution is arranged in the shaking bottles, two adjacent shaking bottles are connected through a functional siphon guide tube solid culture connector, and finally the shaking bottles are sequentially connected in series, the functional siphon guide tube solid culture connector comprises a siphon guide tube, two ends of the siphon guide tube are provided with needles which are respectively inserted into the two adjacent shaking bottles and are not in contact with the liquid level of the culture solution, an adsorption carrier and a solid culture medium are arranged in the siphon guide tube, only a target strain is added into the first shaking bottle, solid-liquid alternate culture is realized through oscillation of the material-supplementing shaking table, and a product is obtained in the last shaking bottle.

In the above technical scheme, the siphon catheter can adopt silicone tube, medical plastic tube, stainless steel tube, glass tube, etc.; the inner diameter is usually 0.25-5.0 mm, the length of the pipeline is usually 30-300 mm, needles used at two ends of the catheter are 8-20-gauge needles, and the length of the needle handle is 25-150 mm; the adsorption carrier material in the siphon catheter comprises cotton thread, absorbent cotton, nano artificial fiber, absorbent gel, agar and the like; but the materials, the pipe diameter, the length of the pipe, the size of the needle head, the distribution system of the carrier components in the pipe and the like used by the functional siphon pipe have key influences on the culture of target strains, so the siphon pipe can be specially designed according to different use purposes;

when the method is used for domesticating and separating the high-activity pectinase aspergillus niger, the siphon guide pipe adopts a silicone tube with the inner diameter of 6-8 mm and the length of 60 mm; absorbent cotton is selected as a carrier material for adsorption in the tube, an agar nutrient medium is adopted as a solid culture medium in the conduit, nutrient substances are consistent with culture solution in a corresponding upstream bottle, a syringe needle with the inner diameter of 0.8mm is selected as the needle head, and the length of the needle handle is 50-100 mm; and arranging culture solution in each shake flask, wherein the distance between a needle and the liquid level is 2-3cm, the concentration of glucose in the culture solution in each shake flask from the first shake flask is gradually reduced, the content of pectin powder is gradually increased, after the whole shake flasks are sterilized, adding a target strain Aspergillus niger A26 into the first shake flask, carrying out shaking culture at 23-25 ℃, and obtaining high-activity pectinase Aspergillus niger in the last shake flask.

When the method is used for rejuvenation of beauveria bassiana strains, the siphon guide pipe is a transparent plastic pipe with the inner diameter of 6-8 mm, and the length of the guide pipe is 100 mm; 2% of agar is selected as an in-tube adsorption carrier material, a solid culture medium is consistent with a corresponding culture solution in an upstream shake flask, and 1/2 tube space is reserved when a guide tube is filled so as to facilitate conidium generation; the needle head is an injector needle head with the inner diameter of 0.45mm, and the length of the needle handle is 20-30 mm; and (3) setting culture solution in each shake flask, adding the bollworm larva extracting solution from the second shake flask, increasing the adding amount of each flask, after integral sterilization, adding the beauveria bassiana strain into the first shake flask, carrying out oscillation culture at 25-28 ℃, and finally obtaining the rejuvenated beauveria bassiana strain in the last shake flask.

In addition, the feeding table of the present invention is a ZWYB-292 feeding table from Shanghai Zhicheng analytical instrument manufacturing Co. The device is a limit 6-bottle fermentation, double-display large-screen touch control panel, closed-loop micro pulse accurate material supplement, and keeps the biological culture environment in an optimal control state; PH control, nutrition supplement, temperature and rotating speed are continuously adjustable; wherein the PH control and the nutrition supplement are respectively controlled by a micro electromagnetic pump, and the precision reaches micro-millilitre level; the adopted double-display large-screen touch controller realizes man-machine operation, is visual and convenient, displays six PH values and growth corresponding curves and trace acid regulating process curves of 6 shake flask reactors in real time, and has advanced functions of online observation, recording, storage, comparative analysis, remote data output and the like in the fermentation process (as shown in figure 1).

The invention has the advantages that:

1) the strain domestication operation process is simplified, and the prior strain domestication is still finished by liquid culture after a target strain is obtained by plate solid culture and selection and is repeatedly and alternately finished for multiple times. If a certain strain of mould needs 72 hours for acclimatization and fermentation each time, the acclimatization and culture are carried out for 6 times, the strain is transferred to solid culture after liquid fermentation and culture, then the strain is selected to enter the next liquid culture, and the time for repeatedly and completely completing the process is at least four weeks or even longer; the siphon catheter connector technology is adopted, firstly, the type of the catheter material is selected, the diameter and the length of the catheter with the adsorption carrier material in the catheter and the model of a catheter needle are determined, each section of catheter is placed into the set solid culture medium, domestication and culture are carried out for 6 times, only 1 time is needed for strain transfer, and only 1-2 weeks is needed for apparent adaptive domestication.

2) The siphon catheter solid culture connector can be manufactured into a standard biochemical kit, and the standard biochemical kit is stored in a sealed mode, can be repeatedly operated, reduces artificial experimental errors, and saves a large amount of experimental equipment.

3) A functional siphon conduit material supplementing shaking table is characterized in that a plurality of fermentation bottles are communicated after a solid culture medium is arranged in a conduit, so that the prior solid-liquid alternative cultivation and fusion are completed on the shaking table at one time, and a siphon conduit is a great technical improvement on the application of the material supplementing shaking table, brings a new market development space for the development and application of products developed by the shaking table, and is suitable for being widely applied to the fields of food, medicine, feed, biological products and the like.

In summary, the innovation of the invention is as follows:

1) through a siphon conduit solid culture connector, strains are domesticated, separated and rejuvenated by solid liquid alternately and respectively cultured, so that the strains are simply integrated on a shaking table, and are inoculated once and continuously fermented for multiple times until a high-quality product is obtained;

2) the Darwin biological evolution theory is used as a guide basis to 'environment selection adaptive survival', a solid culture medium in a siphon conduit is strictly arranged, and the siphon conduit is used as a conduit length, a conduit material, a needle head aperture size, an in-conduit culture medium carrier preparation and the like of a connector to select and adapt related keys for domesticating and culturing microorganisms such as mould, bacteria, saccharomycetes, actinomycetes, viruses and the like, so that the siphon conduit has a very wide application field.

3) The functional siphon catheter connector is used as a new strain breeding technology, is made into a standardized preparation box which can store massive biotechnological information and is convenient for universal exchange of strain resources, so that good strain domestication, separation and rejuvenation depend on not only the strain, but also which kind of feeding table siphon catheter connector is adopted, the application effect of an analysis table is greatly improved, and the technical blank that the feeding table analysis table is a product development application table is filled.

Drawings

FIG. 1 is a schematic view of the structure of a feeder-shaker;

FIG. 2 is a schematic diagram of the process of the present invention;

FIG. 3 is a schematic diagram of the connection of siphon pipes between the flasks;

FIG. 4 is a schematic diagram of the structure of a functional siphon conduit solid culture connector.

In the figure: 1. the device comprises a ZWYB-292 type material supplementing shaking table chassis, 2 material supplementing triangular shaking bottles I-VI, 3 nutrition infusion bottles I-VI, 4 pH meters I-VI, 5, input alkali liquor bottles I-VI, 6 nutrition liquor bottles, 7 alkali solution bottles, 8 alkali solution tube pumps I-VI, 9 nutrition liquor tube pumps I-VI, 10 and a precise micro pump; 11. siphon pipes I-II, 12, siphon pipes II-III, 13, siphon pipes III-IV, 14, siphon pipes IV-V, 15, siphon pipes V-VI, 16, a clamping belt type data connecting line, 17 and a double-line touch screen.

Detailed Description

The method of the present invention is further illustrated with reference to the accompanying drawings and examples.

The invention relates to a siphon conduit feeding shaking table culture method for bacteria, which is based on a feeding shaking table, wherein the feeding shaking table comprises a plurality of shaking bottles, culture solution is arranged in the shaking bottles, two adjacent shaking bottles are connected by a functional siphon conduit solid culture connector, and finally the shaking bottles are sequentially connected in series, the functional siphon conduit solid culture connector comprises a siphon conduit, two ends of the siphon conduit are provided with needles which are respectively inserted into the two adjacent shaking bottles and are not contacted with the liquid level of the culture solution, an adsorption carrier and a solid culture medium are arranged in the siphon conduit, only a target strain is added into the first shaking bottle, solid-liquid alternate culture is realized by oscillation of the feeding shaking table, and a product is obtained in the last shaking bottle.

The invention can add 11-15 functional siphon catheter solid culture connectors (shown in figure 2 after connection) on the basis of a ZWYB-292 type feeding table (figure 1), the connection method is that one end of a siphon catheter needle is inserted into a rubber cap at the bottle mouth 2-3 of a shake flask 2-I, the other end of the siphon catheter needle is inserted into a rubber cap at the bottle mouth 2-3 of a shake flask 2-II to form a siphon catheter I, and all adjacent shake flasks are sequentially connected to form siphon catheters I-V in the same way. When in use, only strains are added into the shake flask 2-I, the two needles of the siphon pipe solid culture communicating vessel are inserted into the fermentation flask openings 2-3-I-2-3-VI in series to form the alternation of shake flask liquid fermentation and siphon pipe solid fermentation, and finally the product is obtained in the shake flask 2-VI (figure 3).

The examples are as follows:

1. domesticating and separating high-activity pectinase aspergillus niger:

pectinase is a commonly used biological enzyme preparation for extracting juice in the production of juice beverages.

1) Experimental materials: pectinase Aspergillus niger A26 (available from Beijing microorganisms) has a basic pectinase activity of 80 ten thousand units/100 g; nutrient solution: 2.0g/100ml of monopotassium phosphate; magnesium sulfate 0.15g/100 ml; peptone 0.30g/100 ml; the concentration of glucose is 1.5g/100 ml; pectin powder 0.1g/100 ml; the prepared nutrient medium is subpackaged in 6 volumetric flasks of 1000 ml; each containing 300ml of liquid; from 2-II to 2-VI of the shake flask, respectively adjusting the glucose concentration (g/ml) of each bottle of nutrient solution: 1.4%, 1.3%, 1.2%, 1.1%, 1.0%; and simultaneously respectively preparing the pectin powder contents (g/ml): 0.2%, 0.3%, 0.4%, 0.5%, 0.6%; the content of glucose added into the nutrient solution supplement bottle 6 is 1.0g/100ml, and the filling amount is 300 ml; the alkali solution replenishing bottle 7 was charged with 300ml of sodium thiosulfate having a concentration of 1.2g/100ml for adjusting the pH.

2) The instrument comprises the following steps: the model ZWBY-292 feeding table (see FIG. 1) from Shanghai Zhicheng analytical instrument was used; installing siphon catheter connectors (see fig. 2): a silicone tube with the inner diameter of 6-8 mm and the length of 60mm is adopted; absorbent cotton is selected as the adsorbing material in the tube, the nutrient medium in the guide tube is consistent with the nutrient solution in the corresponding bottle, and agar is added for 1.8g/100 mm; the needle head is an injector needle head with the inner diameter of 0.8mm, and the length of the needle handle is 50-100 mm;

3) the operation method comprises the following steps: taking out a ZWYB-292 type feeding table bottom plate 1, assembling six large-capacity (1000ml) fermentation bottles 2, numbering the bottles according to the number of 2-I-2-VI in a clockwise mode, adding prepared nutrient solution (250ml) into the bottles, covering a bottle mouth 2-1 with cotton, covering a bottle mouth 2-2 with a rubber cap, sealing, inserting a PH meter detection electrode 4 and an alkaline solution feeding pipe needle 5, connecting the other end of the alkaline solution feeding pipe to the output end 4-I-4-VI of an alkaline solution feeding precision micro-pump 8, and connecting one corresponding input end pipeline (1 for 6) to an alkaline solution feeding bottle 7; the bottle mouth 2-3 is covered with a rubber cap, the needle heads 3 of the nutrient solution supplementing pipes are respectively inserted into the rubber cap, the other end of each nutrient solution supplementing pipe is connected with the output ends 3-I-3-VI of the nutrient solution supplementing precise micro pumps 9, and one corresponding input end pipeline (1 supply 6) is connected with the nutrient solution supplementing bottle 6; and (3) inserting siphon guide pipe communicating vessels into each bottle opening 2-3 in pairs after connection according to the above, forming connecting siphon guide pipes I-V, enabling the distance between a needle head and the liquid level to be 2-3cm, finally integrally sterilizing the bottom plate 1, cooling, adding a target strain Aspergillus niger A26 into the shake flask 2-I under an aseptic condition, integrally mounting the bottom plate 1 on a material supplementing shaking table, controlling the temperature in the shaking table to be 23-25 ℃, and carrying out shaking culture. According to ZWBY-292 type fed-batch shaker detection, target strains added in 2-I after shaking culture for 72-96 hours begin to recover, liquid shaking adaptation survival strains are adsorbed into a siphon guide pipe I to enter a solid culture stage, newly-grown aspergillus niger strains are subjected to shaking dip-dyeing by nutrient solution of a shake flask 2-II at the other end of the siphon guide pipe I after the solid culture is mature, and the living newly-grown strains are propagated and amplified in the shake flask 2-II through selection adaptation, so that the continuously alternate fermentation culture is carried out until the shake flask 2-VI obtains the low content of utilized glucose, the high content of utilized pectin powder, the newly-evolved adaptation aspergillus niger strains, and the pectinase activity is more than 80 ten thousand units/100 g. The strong utilization capacity of the strain on the pectin powder shows that the content of the pectinase is high.

2. Rejuvenation of beauveria bassiana strains

The biological pesticide is an ideal plant protection preparation pursued by organic agriculture, has specific killing effect on pests, and is harmless to human, livestock and environment. Beauveria bassiana is taken as an example, and the Beauveria bassiana belongs to the same eukaryotic filamentous fungi as the mould. The beauveria bassiana individuals are divided into three states, namely mycelium, node spores, conidiophores and conidia; in either state, new beauveria bassiana can be derived. Beauveria deuteroides, Pediobolus subdirectories, Pediobolus conidae, Beauveria. Two of these species, the globularia and the ovate muscardine species, are the most used at present in the former species. The beauveria bassiana hyphae is white and transparent, irregular intervals are formed in the middle under a humid environment, so that sections with unequal lengths are formed, new hyphae can be formed after each section is broken, the diameter of the hyphae is usually 3.5 mu m, and the length of spores forming the sections is different from 3.0 to 5.0 mu m; by utilizing the characteristic, the industrial biological pesticide can be prepared by liquid fermentation. Conidiophores and conidia can be formed under a certain temperature and humidity, the conidia of the beauveria bassiana are spherical, the diameter of the conidia is 2-3 mu m, the beauveria bassiana is in the minimum state, and the host is most easily infected. The principle of the beauveria bassiana biological insecticide is as follows: the beauveria bassiana is easy to be immersed into lepidoptera, homoptera and hymenoptera insect larvae in the smallest individual form, taking lepidoptera noctuidae cotton bollworms as an example, the larvas with large sizes of 5-6 days old enter the feeding burst period, and the body forms are as follows: diameter × length ═ 4.3 × 50 mm; the abdomen of the cotton bollworm larva is provided with a plurality of rows of vent holes, the diameter of each vent hole is 0.025-0.045 mm along with the size of the body, and the vent holes are dozens of times of the spherical beauveria spores. The beauveria bassiana biological insecticide is generally prepared into powder to be poured and scattered on the surfaces of plant leaves, flower catkins or all corners of fields, beauveria bassiana spores are wrapped on the abdomen as long as cotton bollworms crawl through peristalsis, the beauveria bassiana spores enter the bodies of cotton bollworm larvae after peristalsis and breath opening, the body fluid of the larvae is utilized to rapidly propagate to form mycelia, the mycelia are broken into nodose spores, then more mycelia are formed, and the larva organs are made to decay and die due to stiffness until the mycelia are full of larva body cavities. The larva body skin can burst after hyphae are filled in the larva body cavity, and the hyphae can rapidly grow out of the larva body to form conidiophores and a large number of conidia, so that obvious white villi are formed on the body surface of the larva killed by the beauveria bassiana. The newly born large amount of conidia are diffused to each corner of the field under the action of wind vector, so that more larvae can be infected, and the infected stiff insects fall into the soil of the field, so that the beauveria bassiana is protected; thus, biopesticides have an insecticidal effect once in every year, and the use of biopesticides outside the used area can cause a local outbreak of cotton bollworm blight, which is an advantage that chemical insect killers cannot be compared with. The quality key of the biological insecticide lies in the activity of strains, and the strain activity is reduced due to improper strain preservation process or excessive strain transfer, so that the expression staining ability is reduced, and the original staining ability is reduced to 23% or lower from 100%. The best method is to utilize parasitic pests to prepare extracting solution to culture the beauveria bassiana for multiple times, and improve the specific parasitic capacity and parasitic sensitivity of the beauveria bassiana to the pests.

The specific method comprises the following steps: 1) materials: beauveria bassiana strain (sold by Beijing microorganisms); the extract of the cotton bollworm larvae is prepared by taking 10 g of the living cotton bollworm larvae of 5 days old, crushing the living cotton bollworm larvae by using a cell grinder, adding sterile water for dilution and filtration, collecting coarse residues, grinding again, extracting filtrate to a constant volume of 100mL, and keeping the integrity of the components containing the cotton bollworm in the filtrate as much as possible.

2) Preparing nutrient solution (g/ml): 3% of peanut cake, 3% of cane sugar, 0.02% of monopotassium phosphate and 0.01% of magnesium sulfate; the above are packed into 6 volumetric flasks of 1000ml, each containing 200ml of the nutrient solution; wherein, 0.2g of peptone is added into 2-I of the shake flask, and the crushing extract of the cotton bollworm is respectively added into 2-II to 2-V of the shake flask: 5, 10, 15, 20, 25 ml; 300ml of the preparation solution is added into the nutrient solution supplementing bottle 6, and 25ml of the cotton bollworm crushed extract is added. An alkali solution replenishing bottle 7 was charged with 2.0g/100ml of sodium thiosulfate and 300 ml. The size and the growth characteristics of conidium volume of beauveria bassiana are shown, the siphon catheter connector catheter adopts a transparent plastic tube with the inner diameter of 6-8 mm, and the lengths of the catheters I-V are all 100 mm; 2% of agar is selected as an adsorbing material in the tube, the nutrient medium is consistent with the nutrient liquid in the corresponding shake flask, and 1/2 tube space is reserved when the catheter is filled, so that conidia can be generated conveniently; the needle head is an injector needle head with the inner diameter of 0.45mm, and the length of the needle handle is 20-30 mm.

3) The operation method comprises the following steps: the ZWYB-292 type feeding table manufactured by Shanghai Zhicheng analytical instrument manufacturing Limited is selected as the instrument, and is specifically connected with the feeding table in the embodiment 1; and finally, sterilizing the whole base plate 1, cooling, adding a small amount of Beauveria bassiana strains into the shake flask 2-I under an aseptic condition, mounting the whole base plate 1 on a feeding material shaking table (shown in figure 2), controlling the temperature to be 25-28 ℃, and carrying out shaking culture. Detection shows that after 8-10 hours of shake culture, the added beauveria bassiana strain in the shake flask 2-I begins to recover, liquid is selected by oscillation, the adapted survival strain is adsorbed into the siphon catheter I to enter a solid culture stage, a new beauveria bassiana strain is soaked and stained by the shake flask 2-II nutrient solution at the other end of the connector after the solid culture is mature, the new beauveria bassiana strain adapted to survive is selected again to propagate and amplify in the shake flask 2-II, the ZWBY-292 type feeding table is used for monitoring feeding and adjusting PH in time, and the materials are continuously and alternately fermented and cultured in the shake flask 2-VI in such a way, and finally, the beauveria bassiana strain which is highly sensitive after rejuvenation and has extremely strong parasitic cotton bollworm activity is obtained.

The rejuvenation of the strains can not be finished only by using a feeding table, and the operation is complicated and difficult if solid culture is completely used. The siphon conduit solid culture connector is adopted to realize the alternate fermentation of the solid liquid, which is a new function of product development and application endowed to the feeding analysis shaking table; particularly, nutrient substances in the catheter are strictly prepared, produced in batches and stored in a sealed mode, repeated production of the shaking table can be achieved, and an important technical guarantee is provided for standardized production of high-quality strains.

Claims

1. A siphon pipe material-supplementing shaking table culture method for mould is characterized in that a material-supplementing shaking table is taken as a basis, the material-supplementing shaking table comprises a plurality of shaking bottles, culture solution is arranged in the shaking bottles, two adjacent shaking bottles are connected through a functional siphon pipe solid culture connector, the shaking bottles are sequentially connected in series finally, the functional siphon pipe solid culture connector comprises a siphon pipe, needle heads are arranged at two ends of the siphon pipe and are respectively inserted into the two adjacent shaking bottles, the distance between the needle heads and the liquid level of the culture solution is 2-3cm, an adsorption carrier and a solid culture medium are arranged in the siphon pipe, a target strain is only added into the first shaking bottle, solid-liquid alternate culture is realized through oscillation of the material-supplementing shaking table, and a product is obtained in the last shaking bottle.

2. A siphon tube feeding and shaking table cultivation method for mold as claimed in claim 1, which is used for domesticating and separating high activity pectinase aspergillus niger, wherein the siphon tube adopts a silicone tube with an inner diameter of 6-8 mm and a length of 60 mm; absorbent cotton is selected as an adsorption carrier in the tube, an agar nutrient medium is adopted as a solid culture medium in the conduit, nutrient substances are consistent with culture solution in a corresponding upstream bottle, an injector needle with the inner diameter of 0.8mm is selected as a needle head, and the length of a needle handle is 50-100 mm; and arranging culture solution in each shake flask, gradually reducing the concentration of glucose in the culture solution in each shake flask from the first shake flask, gradually increasing the content of pectin powder, after the whole shake flasks are sterilized, adding a target strain Aspergillus niger A26 into the first shake flask, carrying out shaking culture at 23-25 ℃, and obtaining the high-activity pectinase Aspergillus niger in the last shake flask.

3. A siphon tube feeding and shaking table cultivation method for mold as claimed in claim 1, wherein for rejuvenation of beauveria species, the siphon tube is made of transparent plastic tube with inner diameter of 6-8 mm, and the length of the tube is 100 mm; 2% of agar is selected as the adsorption carrier in the tube, the solid culture medium is consistent with the culture solution in the corresponding upstream shake flask, and the space in the tube of 1/2 is reserved when the catheter is perfused, so that conidia can be generated conveniently; the needle head is an injector needle head with the inner diameter of 0.45mm, and the length of the needle handle is 20-30 mm; and (3) setting culture solution in each shake flask, adding the bollworm larva extracting solution from the second shake flask, increasing the adding amount of each flask, after integral sterilization, adding the beauveria bassiana strain into the first shake flask, carrying out oscillation culture at 25-28 ℃, and finally obtaining the rejuvenated beauveria bassiana strain in the last shake flask.