TWI338048B - - Google Patents

Description

1338048 IX. Description of the Invention: [Technical Field] The present invention relates to a method for producing a fungichromin in a large amount of liquid culture, which is a liquid culture Sirepio melon/as PMS-702 in a fermentation tank. Generate a mildew base. [Prior Art] In recent years, due to the rising awareness of environmental protection, the concept of reducing the use of chemical agents and changing the cultivation of crops by organic farming methods has been paid more attention to. Organically, organic crops can be used to reduce the chemistry of the people. The pollution of pharmacy and fertilizers can achieve the function of maintaining ecological balance, saving resources available in the natural world, and making the land sustainable. Among them, the beneficial bacteria in the soil cockroaches are screened by the bio-separation method, and the microbial agent is used to replace the chemical agent, which is a method for controlling pests and diseases by using microorganisms. Can't rely on the speed of reproduction to compete Some microbes grow at a slower rate of survival resources, but must be manufactured by chemicals that inhibit the growth of other organisms. This record. Ink 铷 从 from the pill > /

Available in extremely dilute concentrations - functional. The anti-obstructive metabol i te ), which is not a raw synthesis in pure culture, requires a very complex metabolism, and in the case of a decrease in growth rate, the gene involved in the synthesis of microorganisms synthesizes related enzymes and once metabolites. For example, there are many types of microorganisms that have the ability to produce antibiotics. 5 B38048 letters and actinomycetes, among which, the germline is the genus of chain bacterium (10) (4) Qing). Streptomyces belongs to actinomycetes, which is a saprophytic bacterium, which is ubiquitous in root zone soils. It is the main source of antibiotics. Many important antibiotics (actin(10)yc:ln), sulphate (... mail (10) ycin ), tetracycline, pQlyQxin, kaSUgamycin, Pimaricin, and kangmycin can all be produced by Streptomyces. Streptomyces has good production properties of extra-column hydrolyzing enzymes, which can be used to decompose various biopolymers such as chitin, cellulose and 1'3 glucomannan present in the natural environment. Molds can still produce spores that have good resistance to environmental stress. With the above characteristics, Streptomyces is considered to be a good species that is well suited for development as a biological control agent. In the various aspects of biochemical process development, including the upstream strain improvement screening, the midstream fermentation process development to the downstream product purification and recovery, each link will affect the success or failure of the overall process, which is the development of acid fermentation process and mass production amplification. More important and indispensable ring. 57 factory printing ^ fresh £^5; 73 office / 7 " 15? ^ ^ - 702 is a strain with the ability to produce antifungal active antibiotics, the strain is obtained from field soil, compost and cultivation medium in Taiwan Among the 200 strains of actinomycetes, they were screened and obtained as 'strained by 4 strains, and named as calendar/ce>5 PMS-702. The antibiotics produced by this strain were analyzed by infrared spectroscopy and nuclear magnetic resonance spectroscopy. And the mass spectrogram analysis confirmed that it is a polymorphic macrolide of the polyene macrolide (f ungi chromi η). The antibiotic has an inhibitory effect against Rhizoctonia solani, 6 5 13*38048 and it has a complete inhibitory effect on the release of cysts and migratory spores of Phytophthora infestans, and for the 5Y plant, "" The fermentation broth was subjected to bioassay using plants, fruits or cut leaves, and found to have brown spot disease of lettuce, anthracnose of cabbage, mango anthracnose, citrus green mold, cabbage bacillus and peach fruit blight. According to the control effect, vSYrepi(4) pMS_7〇2 is a strain with the potential to develop a plant protection preparation (Sh iheia A, 2003) a Therefore, it is an important subject to explore its culture conditions. In the previous study, the best medium formula (S, 2004) was obtained in s pms_7〇2 shake flask culture, but it was cultured in a fermentation tank. The condition has not been established, so that f ungichromin can not assist the biological control work by the large-scale manufacturing. SUMMARY OF THE INVENTION The optimal conditions for the production of mycelium-based cells by id in the liquid culture of the fermentation tank are not established. Therefore, the present invention proposes a fermentation tank culture condition, in order to effectively improve the mildew treatment. The yield of color bases makes it widely applicable to biological control work. The main object of the present invention is to provide a method for producing a mycophenolic substance in a large amount of liquid culture. The technical means for achieving the above purpose is to add about 0_01 to 2·% defatted soy flour, about 1 to 20% glucose, and about 0.01 to 1.0% calcium carbonate in the fermentation tank. For the culture medium, fresh ces PMS-702 was cultured in the above culture medium, 13*38048 and the culture conditions were set to a rotation speed of about 10 〇 to 4 ο rpm, and the ventilation was about 5 5 to 1 · 5 vvm and the culture medium. It is weakly acidic for cultivation. Preferably, the rotational speed at the time of culture is set to about 2 Torr to 35 rpm. Preferably, the rotational speed at the time of culture is set to about 3 rpm. Preferably, the ventilation during culture is set to about 〇 8 to! 2vvm. Preferably, the amount of ventilation during culturing is set to about i 〇 vvm. Preferably, it further comprises adding a divalent magnesium ion, a divalent zinc ion, a divalent copper ion, a divalent iron ion or an inorganic phosphate, or a combination thereof, at the time of culture. Preferably, it is a bivalent magnesium ion of about _0_5 to i0_2M added during the culture. The car is good again!!: 纟 培养 培养 培养 培养 培养 培养 1 培养 培养 培养 培养! 〇 _ 5 pairs of the second price recorded ions. Preferably, it is a dying zinc ion of about ΗΓβ 1G 2 M when it is cultured.

Preferably, copper ions are preferred. Preferably, iron ions are preferred. It is added when it is cultured, and it is added when it is cultured. It is added when it is cultured. It is added about 1% of the divalent zinc ion when it is cultured. 1 〇-5 to 1 (2% of KM) The divalent copper ion of Γ4Μ. 10 -5 to 1 (the valence of Γ3Μ is preferably an acid salt. It is added in the culture σ. It is added about 10_4 Μ of the ferrous ion in the culture. Preferably, the inorganic phosphorus is about 1 〇 4 to 1 〇 2 Μ, and it is about 6 〇 3 Μ Μ Μ Μ Μ 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 13 13 After the time, the oil is added to the feed. Preferably, the oil to be added is one or more selected from the group consisting of big five oil, cottonseed oil, corn oil, eucalyptus oil and oleic acid. Preferably, the oil is added to corn oil. Preferably, the oil is added to about 33% (w/w) corn oil. By the above method, a large amount of heart Γ PMS can be cultured in the fermentation tank. -702 for the production of mycelium-based compounds, and the application of mycophenolic compounds for biological control purposes. [Embodiment] The main purpose of the present invention The effect of different culture conditions on the production of fungichroniin in Sirepio/p/ces PMS-7〇2 was investigated in the liquid culture in the fermentation tank, and the application of a large amount of liquid culture to produce mildew-like substances was obtained. The best method's in order to mass produce the mildew-based element 0

When the liquid culture is carried out in the fermentation deduction, the various environments in the fermentation tank are: the growth of the strain, the nutrient utilization during growth, and the metabolite yield of the strain: 'The invention is changed by setting different operations. t, such as rotational speed, ventilation and pH, etc., to understand the environment most suitable for the use of yttrium; 2, the invention is: after the good environment, the feeding additives, including Adding sugar, ions or oils, because the feed of the cultured feed is also added to the fermentation tank environment of 1338048 bases, and further combined with the most appropriate feed, the mold base can be higher. The output is conducive to industrial use. Other features and advantages of the present invention will be apparent from the following detailed description of the invention. EXAMPLES The following examples are presented to illustrate the invention. The examples are not intended to limit the scope of the invention in any way, but are intended to indicate how to practice the materials and methods of the invention. Example 1 Fermentation tank batch type liquid fermentation 1.1 material The strain used in the invention is fresh ces PMS- 702 ' is provided by the Disease Management Laboratory of the Department of Plant Pathology of Zhongxing University, and the strain is produced by one strain. A strain of an antifungal active antibiotic which can be produced by fermentation to produce an antibiotic virgin chromin. This strain was deposited with the Food Industry Development Institute on October 24, 2001 under the accession number CCRC910179. In the following tests, 472·472% defatted soy flour, 23.23% glucose (glucose) and 〇063% calcium carbonate (CaC〇3) were used as the culture medium and cultured in 5L·fermentation tank. . H2 different rotation speeds for 5*. The effect of PMS-702 on the fermentation of batches in the fermentation tank. The purpose of the fermentation is to distribute the matrix required for oxygen and microbial cell growth and to break up the bubbles to increase Dissolve oxygen and avoid the occurrence of cell 3 1338048 agglomeration' and promote the efficiency of mass and heat transfer. In this embodiment, the rotation speed of the tank is changed to discuss the most suitable stirring rate, and other culture conditions are set to a working volume of 3 L, a temperature of 23.8 t:, aeration volume of 5.5 vvm, and pH is not controlled, and the stirring speed is set to 1 〇〇, 2〇〇, 30 0 and 400 rpm, the test results are as shown in the first figure in the 'four stirring speed conditions' with the speed of 300 rpm as the most suitable condition. Under these conditions, after 3 days of brewing Fermentation time can make the yield of fungichromin reach 180.8mg/L. When observing the effect of different rotational speeds on the dissolved oxygen content, it was found that the dissolved oxygen provided by the rotational speed of 1 〇〇 rpm was obviously insufficient. 'Because the fork PMS-702 is an aerobic microorganism, the lack of solubility can lead to poor glucose utilization. The production is reduced; as the stirring rate is increased and the dissolved oxygen state is improved, the cells grow rapidly. When the rotational speed reaches 400 rpm, although the dissolved oxygen is in good condition, high shear stress is generated at this time, which leads to limited growth of the cells, and higher rotational speed causes more hyphae to adhere to the upper part of the groove wall, reducing The mycelium is in contact with the φ liquid medium, thereby reducing the proliferation of the mycelium and reducing the yield of fungichromin. From the above, it can be seen that the optimum speed of the sputum s pMS_7 〇 2 in the fermentation tank is 3 0 0 r p m. ^丨.3 Different aerations for 乂P5; pMS_7〇2 in the fermentation tank, the effect of batch fermentation, oxygen can supply the growth of bacteria, and the demand for aerobic microorganisms will change continuously during the growth period. The production of the product and the maintenance of cell metabolism, (Oxygen requirement) in the absence of oxygen supply, will affect the growth or product formation of the bacteria 1338048. However, when the dissolved oxygen value is increased, the growth rate of the cells can be increased. The cells become unstable and reduce the production efficiency of the fermentation products. Therefore, it is necessary to investigate the effect of ventilation on the batch fermentation of pms_7〇2 in the fermentation tank to obtain the best conditions. The conditions were set to a working volume of 3 L, a starting pH of 5.5, a collection of 100 mL/L, a temperature of 23.8 ° C, and a rotational speed of 300 rpm, and the aeration conditions were set to 〇.5 vvm, l.〇vvm and 1.5. Vvm. From the results of the beta test, it was found that the dissolved oxygen value increased with the increase of ventilation, and the supply of oxygen was optimal at 1.5 vvm, but when the glucose consumption was observed, as shown in the first figure, the ventilation was found. 1. At 〇vvm, glucose is used up in the 3.5th to 4.5th days. When the ventilation is increased to 15vvm, the glucose run-out time is prolonged (Day 5), which indicates that the high dissolved oxygen amount causes the cells to be caused. The inhibition phenomenon. Referring to the third figure, it can be found that the mycorrhizal production of pupa can be found under the condition of aeration of 1vvm for 4.5 days, and the best yield of mycorrhizin can be obtained. The value is 231.27. Mg/L, verification of the month 1J recital due to excessive ventilation caused by poor production speculation. From the above discussion, it was found that the ventilation was 丨.0vvm, which is the best ventilation culture condition. 1.4 Controlling the effects of different pH values on the batch fermentation of the parent pMS_7〇2 in the fermentation tank 3 The growth and production of microorganisms are closely related to the pH value in the culture environment i appropriate pH regulation strategy for the product The level of production and the length of the bacteria are extremely important factors. In the above discussion of the rotational speed and aeration, the pH value was compared with the pH value of the vinegar fermentation conditions, and the yield of the glutinous 8 1338048 color base was found to be 'yield when the pH was 5 to 7. Preferably, this is the case. P injury test ίτ. In the vinegar fermentation process, the pH value is maintained at 5, 6 and ^ other culture conditions are set to the servant volume 3L, inoculation W / L, dish 23. 8 C, 300 rpm and ventilation Quantities ^ 〇w Referring to the fourth figure, when the pH is regulated at 5, there is a high yield of the color of the chromophore, and when the pH is adjusted to neutrality, the production of the mycelium is also strong. There is inhibition; when observing the utilization of glucose, it is found that pH PH is 6 or 7 is better, but when the amount of dissolved oxygen is compared, pH=7 has higher ritual consumption ϋ pH = 7 when the growth of bacteria Preferably, but the case of producing mildew-like base at this time is not ideal, so (4) pH neutral movement, which is beneficial to the growth of PMS-7〇2 cells, but is not conducive to the production of mildew. It can also be seen from the fourth figure that when the result of not regulating the pH value is superior to the regulation, it is known that the regulation of the pH value is not conducive to the production of the methicone. Example 2 Feed Batch Fermentation in Fermentation Tanks According to the results of the above Example 1, the glucose system was established on the fourth day under the operating conditions of a rotation speed of 300 rPm, aeration amount h〇vvm and no pH control. (β according to the first figure), at this time the pH value moves to the alkaline, showing the bacteria due to •. There is no carbon source available in the nutrient base, and it tends to decompose and die, and this part of the test is to explore the effect of regulating the carbon source on the fermentation. In the following tests, Snapdragon was added to the culture medium, 47 to get s ugly, I. 23% glucose and 〇. 〇63% calcium carbonate, and 5L 酦 slot pairs?丨PMS-702 was cultured; other culture conditions were set to 13 1338048 p value 5.5, inoculum volume i〇〇mL/L, temperature aeration Ι.Οννπι. The working volume is 3L, the initial 23.8 ° C, the rotation speed is 300 rpm, and the pac/a/7i/s· PMS-702 2 · 2 investigates the effect of the glucose feed once added to the fermentation tank. In the second picture and the flute-two diagram, when the concentration of glucose in the medium is lower than 4 g/L, ς". If PMS-702 is used, it will no longer be produced. > Balance: color base; now feed料方戎^ Shanyi supplemented slaves’, after 2 days of cultivation,

The glucose concentration is reduced to about 6 g/i 0 士 ^ , Λ 0 4 'feeds 100 g / L glucose 〇 1 L, so that the sugar concentration is increased to 1 〇 g / L, i step A | - g then M culture On the 7th day, after the primary feeding of the Portuguese syrup, the bacteriological sputum was discussed. . After the test, the results showed that after the glucose was fed on the second day, the dissolved oxygen value was maintained at about 〇 and compared with the glucose without feeding, it was found that the bacteria had sufficient carbon source available after feeding the glucose. Therefore, it is not until 帛5 5 days that the death decomposition begins. Referring to the fifth figure, when observing the yield of the chromophore, it can be found that the serotonin production of $2 after feeding the glucose for 2 days is due to the dilution effect and then shows an upward trend. The maximum value was reached at 4·5 days (208. 67 mg/L). Compared with the fermentation condition without glucose feed, feeding glucose into the material did not increase the yield of the mycelium. When the field feeds a large amount of glucose, the bacteria will use it for growth, but will not use it in the growth path of secondary metabolites (such as antibiotics). 仏3仏^glucose continuous feed addition pair & /750 ^/7"$ PMS-702 The effect of batch fermentation in the fermentation tank is known as the method of continuous feeding of glucose, sampling every 8 small 14 3 and analyzing the knots (the first) Two days), that is, hold: Second, when the glucose concentration is found to be lower than 6 4 ff / I 0. more than g / L, meaning that when the concentration of carbon in the matrix is lower than feeding the appropriate amount of glucose solution, the sugar concentration is maintained at 4 g/L or more, continue to train until 14 hours, when you are under the batch, keep the Portuguese exhibition and depreciate Its impact on the growth of bacteria and mildew treatment Kissin production of color. , seat. The test results show that due to the continuous feeding of the carbon source, the bacteria have sufficient carbon source, and the phenomenon of "hyperactivity" to the alkaline PH (four) is not found even after 40 hours of culture. . In addition, as can be seen from the sixth figure, although glucose is continuously fed into the medium, there is no improvement in the production of the mycorrhizal substance by the parent. In Example 2. 1 and 2.2, neither of the mycotoxins increased the yield due to the supply of glucose. The reason is that antibiotics belong to the secondary metabolites of microorganisms, and the nutrient deficiency and growth rate decrease. In this case, the microorganisms will begin to express the genes related to the secondary metabolic pathway and the biosynthesis-related enzymes to facilitate the production of a metabolite. Therefore, PMS-702 will produce a mildew-based substance in a nutrient-rich environment. Limited, so that its production cannot be improved. Example 3 Discussion of shake flask additives This example is to investigate the effect of different ionic additives and oil additives on the production of mycelium-based compounds of S PMS-702. 3. 1 Materials In this example, iron (Fe2+), magnesium (Mg2+), zinc (Ζη2+), copper (Cu2+), 1338048 phosphate (P(V_) and ammonium sulfate ((hh4)2so4) are selected as ions. The object of the additive is added in a concentration range of 1〇-5 to 1〇-ιΜ. In this embodiment, soybean oil “cottonseed oil” and corn oil are added as direct addition. The oily sputum's addition concentration is 〇.5% (w/w), l% (w/w) and 2% (w/w). In this embodiment, the following objects are additionally selected as the feed additive: (1) Oleic acid feed. Configuration 〇·1% (W/W), 〇.5% (w/w), i%(w/w) '1.5%(w/w), 2%(w/ w), 2.5% (w/w), 3% (w/w), 3.5% (w/w) oleic acid aqueous solution, sterilized at 121t, 15 minutes. (2) Corn oil feed: configuration i %(w/w), 9%(w/w), 16%(w/w), 33%(w/w), 50%(w/w), 66%(w/w), l〇〇 %(w/w) of corn oil solution was sterilized at 1 2 rc, 15 min. (3) Cottonseed oil feed: configuration i% (w/w), 9% (w/w), 16% (w/w), 33% (w/w), 50% (w/w), 66% (w/w), l〇〇% (w/w) of cottonseed oil aqueous solution, with 12Γ (:, 15 Min condition Sterilization. (4) Big oil feed: 1% (w/w), 16% (w/about, 33% (w/w), 50% (w/w), 66% (w/w) ), l〇〇% (w/w) of soybean oil aqueous solution, sterilized by 1 2 PC, 15 min. 3. 2 The effect of begging ion addition on & pMS 7〇2 in shake flask fermentation

The preferred concentration is 1 (Γ4Μ, this thick

As a result of the yield of alizarin, when the concentration increases the degree of F e 2 + and inhibits the growth of the cells: the amount decreases. The addition of Mg2+ from 1〇-5 to 10_2M 16 8 13*38048 has an effect on the production of the chromophore. Adding a low concentration of zinc ions (1 〇 -3 to 1 〇 -1 ) is beneficial to the growth of the cells, but at this time, the yield of the methicone is lower, and the addition of 1 Ο·3 Μ zinc ions is improved. The effect of the production of mycorrhizin is 'but when the concentration is increased to 1 Ο-%, the formation of the chromophore is completely inhibited, indicating that the addition of a high concentration of zinc ions inhibits the growth of the cells. The addition of Cu2 + at a concentration of 1 〇 _4 μ has the effect of increasing the yield of the basal color. The increase is 10 Μ 3 Μ and the inhibition occurs.

In general, the addition of phosphates is more conducive to the formation of primary metabolites, but inhibits the synthesis of secondary metabolites (such as antibiotics), indicating that the addition of inorganic phosphates generally inhibits the treatment of mycelium. The yield, but more specially, is the effect of increasing the yield of mycelium when added with 10-3 Μ叩. In addition, the concentration of 卯 添加 在 在 5 5 5 在 没有 没有 没有 没有 没有 没有 没有 没有 没有 没有 没有 没有 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 3. 3 铋彳 grease added to the effect of 孓 办 仙 5 5 pMs_7 〇 2 in the shake flask, this part of the test is used in various concentrations of oil, to add directly or feed to the medium' After culturing 2. After a day, explore the relationship between the production of mycelium and the production of mycelium. The test results show that all kinds of oils can promote the S-suppression training s PMS-702 production and treatment. Different types have different, and, "waves, because of the oil type 1 library does not ~ add too much oil, will Lead to oxygen quality: inhibit the growth of the bacteria. Inductive to improve the mildew base, in the day, the mother's degree is: add 〇, 5% (w / w) to 1% (w / w) soybean oil 8 1338048 It can increase the yield of mycelium to about 335. 37 mg / L, add 0 5% (w / w) cottonseed oil to get about 581 49 mg / L, add l% (w / w) of jade The oil is about 606. 83 mg / L 'The results show that 'the more oleic acid content in the oil and fat' is more conducive to the formation of mildew. The added oil is not limited to the oil of the oily oleic acid containing Dong For example, if the oil is removed, it can also be used as a feed additive. In addition, when the oil is added by means of feeding, it is found that a higher yield of the mildew element is found. The test result is: feeding 5 〇% (w / w) Cottonseed oil can increase the yield of mycelium to 444.99 mg/L, and feed 33% (w/w) corn oil to obtain 567.46 mg/L of mycotoxin yield, feeding 〇% ( w/w) soybean oil is 473.88 mg / L The yield of color base, feeding 2% (w/w) oleic acid can obtain the yield of 796.82 mg/L of mycophenolic base. The effect of the addition of fats and oils on the production of mycorrhizin fermentation by the above In the examples, it is known that the addition of ions or fats can achieve the purpose of improving the mildew-like base, and the use of the added grease method can further improve the yield of the mildew-based base. At this stage, the oil alone is added for discussion. It is a feeding method, and considers the cost and application to the fermentation tank and other factors. 'Select corn oil as the object of discussion, and according to the optimum conditions obtained above, enlarge to 5L vinegar fermentation tank to explore Yiyang 472 472 472 472 472 472 472 472 472 472 472 472 472 472 472 472 472 472 472 472 472 472 472 472 472 472 472 472 472 472 472 472 472 472 472 472 472 472 472 472 472 472 472 472 472 472 472 472 472 472 % defatted soy flour, 1.23% Portuguese 5L fermentation tank, and set conditions of 5, inoculation amount 100 mL / L, temperature 23.8 18 1338048 C: rotation speed 3GG rpm and hGvvm conditions, after a hour of culture, Z The flow rate of 5.5mL/min was fed into 150inL of 33% (w/w) corn oil and continued to be used until day 7. 夂2 test Results The test results showed that after 12 hours of culture, the oil was emulsified with the water in the medium due to the feed of corn oil, so that the dissolved oxygen value was:: After the cultivation of the scorpion, the cells began to rapidly metabolize the medium. (4) Sugar (carbon source), which produces organic acid and moves the pH to acidity. In terms of the use of glucose, the exhaustion of glucose is delayed to 5th compared with when the corn oil is not fed. λ (no feed is the fourth day). As shown in the seventh figure, the product of the mildew-based element in the first 15 days of the culture, no obvious yield appeared, after 2 days of the culture, the bacteria began to produce a large amount of mycelium, after 6 days. In culture, the yield of serotonin can reach a yield of about j mg/L, which is about 87 times that of corn oil. In summary, the culture condition of the present embodiment is a good fermentation condition for producing a mildew-like element by using a liquid culture fork to treat ms PMS-702. Conclusion ^ When liquid culture is used to produce mycelium-based pigments, the preferred conditions for the fermentation tank are 30 rpm and aeration (and the pH is not regulated under this condition) (4) after 4 days of culture. The yield of the chromophores can reach 225mg/L. The supplementation of glucose by one feeding or continuous feeding has no obvious improvement effect on the production of mycelium-based system of PMS_7〇2. In the shake flask medium, add 1 Ο- % of Mg2+, 1 ο-% of Zn2+, 19 8 1338048 10 Μ of Cu2 + or ^, H3 to " position < Fe can increase the yield of the chromophore Μ 2 η 2 Add 1 non-organic phosphate It can be mentioned that it is 42 times, and there is no improvement effect when adding SO4. When adding grease to the shake flask medium, it is best to use straight U(W/W) corn oil for common fats, and its output can be increased to 606.83'. When L' is added by feeding method, α 2%u/w) oleic acid is optimally increased to 796.82 mg/L; and feed added 33% (w/w) corn oil is 567.46 mg/ L.

After various investigations, the conditions of rotation speed _rpm, ventilation 1 (), initial pH 5 5 and whole = fermentation pH control are not applied to the 5L vinegar fermentation tank, and the cost is after a period of cultivation. Add 33% (w/w) corn oil by feeding. When cultured to the 6th day, the yield of mycotoxin can be 2052, 39rag/L, which is about 8_87 times that of corn oil. . [Simple description of the diagram] The first figure shows the effect of different speeds on the batch culture of the fermentation tank and the production of the mildew-based element of the 7M PMS-702. The second graph shows the effect of different aerations on glucose utilization in batch fermentation of PMS-702 in fermentation tanks. The third figure shows the effect of different aeration on the production of mildewin in the batch culture of pai/a/jzys PMS-702. The fourth picture shows the effect of different pH values on the production of mycelium-based compounds by batch culture of PMS-702 in fermentation tank. The fifth figure shows the effect of the addition of a single feed of glucosinolate on the yield of glucose and the production of mycelium by fresh C (?5· Ρ3^/7μΡΜ3-702. 20 8 1338048 The sixth figure is the addition of glucose continuous feed. The effect of glucose utilization on PMS-702 and the yield of chromophore. The seventh figure is the addition of 33% (w/w) corn oil to the fermentation tank batch culture LSYrepi (10) / ces PMS-702 production color The influence of the element. [Main component symbol description]

21 1338048 References l.Shih, H.-D. , L, Y.-C. , Hsu, F.-L. and Vanisree Mu 1abaga1, R. , Fungichromin: A Substance from Streptomyces padanus PMS-702 with Inhibi tory Ef fects on Rhizoctonia solani. J. Agric. Food Chem, 2003. 51: p. 9 5 ~ 9 9. 2. Lin Weicheng, Application 5'Guang 7~67^〇历/(^5'/?3(/&lt ;3/7"5'?^18-70 2 Discussion on the optimum conditions for the production of Fungichromin. 2004: National Chung Hsing University • Master of Science in Chemical Engineering.

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Claims (1)

1338048 X. Patent application scope: The method for producing a mildew-like substance, which comprises: providing a medium, comprising about 〇1 to 5. 0% defatted soy flour, about 〇' 01 to 5. 0% glucose and about 〇 V1至1. v% calcium carbonate; The bacterium is cultured in the above culture medium, wherein the culture condition is a rotation speed of about 100 to 4 rpm, and the ventilation is about 〇. 5 to 1. 5vvm: After the culture for a period of time, the feed is added with oil, wherein the oil is soybean oil, cottonseed oil, corn oil, eucalyptus oil, oleic acid or a combination thereof; and the mildew-like substance is obtained from the culture medium. 2. The method of claim 1, wherein the rotational speed of the culture condition is set to be about 2 Torr to 35 rpm. 3. The method of claim 2, wherein the rotation speed of the culture condition is set to about 300 rpm. The ventilating condition of the cultivating condition is set to about 〇. 8 to 1. 2vvm, as described in the above. The method of claim 4, wherein the ventilation condition of the culture condition is set to about 1. 〇vvm. 6. The method according to any one of claims 5 to 5, further comprising adding a divalent magnesium ion, a divalent zinc ion, a divalent steel ion, a divalent iron ion or an inorganic acid to the culture. Salt, a combination of more than $. 7 'A method of claim 6, wherein the method comprises adding a divalent magnesium ion of about 10-5 to 1 〇 2 》 in the culture system. 8. The method according to claim 7 of the patent application, When cultivating 23 338048 1 -------- The announcement is to add about 1 〇, g 1 镇 离子 离子 , , 〇 _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ When cultured, 1 -2 to 1n-2 χ 〇 i i i i i i i i i i i i i i i i i i i i i 时 如 如 如 如 如 如 如 如 如 如 如 如 如 如 如 如 i i 如 i 如 i i i i i i i i i i i i i i i Zinc ion. Adding heat as described in the scope of claim 6 of the patent, which is from the culture of 2 〇 to 1 () 3 Μ of the divalent copper ion. The time is as in the scope of claim 1 The method described in the method of cultivating a divalent copper ion of about 10 Å to about 3 σ, and adding the method according to claim 6 of the patent application to the method of culturing a divalent iron ion of 10 10 to 1 〇 3 μ. The method of cultivating the time is as described in claim 13 of the patent application, which is a ferric ion of 1 1 1 ο Μ 时 时 时 ^ ^ ^ ^ 申 申 申 申 4 4 4 4 4 4 4 The method comprises the steps of: cultivating 1 10 to 1 〇 2 μ of inorganic carbonates. The method of the cultivating time is the method described in the fifteenth item of the patent application, and the method comprises the following: 'Yu Pei', + plus about 10-3 Μ inorganic phosphates. 1 7 ' The method of claim 16, wherein the six-fuel type is corn oil. The addition of 18* is as described in claim 17, wherein the six or two oils are about 33% (w/w). ) Yulai oil. ' XI, ® type: as the next page 24