CN106399117A - Cordyceps jiangxiensis JXPJ0109 strain and application thereof - Google Patents
Cordyceps jiangxiensis JXPJ0109 strain and application thereof Download PDFInfo
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Abstract
The invention discloses a Cordyceps jiangxiensis strain capable of producing the antineoplastic compound jiangxienone and a method for high-yield production of jiangxienone. It is found in the invention that the Cordyceps jiangxiensis JXPJ0109 strain exerts good effect on synthesis of the antineoplastic compound jiangxienone, and ferrous ions can be added to regulate and control high-efficiency fermentation and metabolism of the Cordyceps jiangxiensis JXPJ0109 strain for synthesis of the antineoplastic compound jiangxienone; so the efficiency of synthesis of jiangxienone through fermentation and metabolism of the Cordyceps jiangxiensis JXPJ0109 strain, and the output of jiangxienone is greatly increased, from 1.0 [mu]g of jiangxienone per g of dried mycelia of the strain to 221.26 +/- 6.79 g of jiangxienone per g of dried mycelia of the strain. The method is simple, easy to operate and especially applicable to deep fermentation of the liquid of the Cordyceps jiangxiensis JXPJ0109 strain for production of jiangxienone; and the method is of important theoretical significance and application value to large-scale production of jiangxienone and research on druggability of jiangxienone.
Description
Technical field
The present invention relates to pharmaceutical field, particularly a kind of Jiangxi Cordyceps Militaris Cordyceps jiangxiensis JXPJ0109 bacterial strain and its application.
Background technology
Cordyceps Militaris are class specific type fungus resources.In long-term evolutionary process, for overcoming the natural cover for defense of host during invasion, and escape invade after host immunosurveillance, reach and synchronize with history of life of host, be finally completed own cells propagation, differentiation and the whole history of life.In this process, generate to cellular signal transduction, stress protect related, have that chemical constitution is various, biological function is various and active secondary metabolite the features such as effect Mutiple Targets, show the potentiality of huge Development of New Drugs.The prompting of existing result of study, have been found that in Cordyceps Militaris rich and varied antitumor activity secondary metabolite (Recent Patents on Biotechnology, 1 (2):123-137,2007；Advance in Biochemical Engineering&Biotechnology,113:79-150,2009).
Jiangxi Cordyceps Militaris Cordyceps jiangxiensis be one novel species of Cordyceps Militaris, be commonly called as " vegetation king " among the people, be treatment venomous snake bite good medicine (Mycosystema, 20:306-309,2003).We, in Cordyceps Militaris screening anti-tumor medicine process, screen a kind of New skeleton compound from the Cordyceps Militaris fermentation mycelium of Jiangxi, are named as jiangxienone (Jiangxi ketene fat).This compound is in different degrees of cellulotoxic effect to the tumor cell line of the separate sources such as humanized's lung cancer, cancer of the stomach, leukaemia, liver cancer, colon cancer, the significant CDCC especially to stomach cancer cell line, it is 6 times of a clinical line cancer therapy drug cis-platinum, but (ZL 201010159145.8 is no affected on humanized's normal cell and mouse bone marrow clone；Chemistry&Biodiversity,9(7):1349-1355,2012；Process Biochemistry,49(4):697-705,2014).Result of study prompting further, the mechanism of action of its suppression growth of tumour cell is related to DNA damage response approach, jiangxienone can embed DNA of tumor cell molecule and cause its double-strand break, thus play neoplasm growth cellulotoxic effect (Process Biochemistry, 49 (4):697-705,2014).Therefore, jiangxienone has the potentiality becoming anti-cancer agent lead compound it would be highly desirable to further it is launched with druggability and preclinical research.And substantial amounts of jiangxienone sample is needed in the development of these R＆D works badly.Under normal condition, the secondary metabolite yield of the microbial metabolism such as fungi synthesis is little, and jiangxienone is only the dry mycelium of 1.0 μ g/g so that the research work in terms of to its follow-up pharmacology, pharmacy is hard to carry in the content in the Chinese caterpillar fungus of Jiangxi.Therefore, need raising Jiangxi Cordyceps Militaris metabolism synthesis jiangxienone yield badly, provide material guarantee for follow-up research and development.
From the forties in last century, using liquid deep layer fermenting by Penicillium chrysogenum producing penicillin since, fungi liquid submerged fermentation technology is ripe day by day.But specific to producing bacterial strain, affect factor and its complexity that its metabolism synthesizes target product.Two big class generally can be classified as, one is the yield lifting target product by fermentation strategies；Two is the biological relations based on target product, transforms bacterial strain using molecular engineering means, improves yield.It is most common for changing fermentation strategies, it is also most efficient method, it involves a wide range of knowledge, the envirment factors such as the medium component (carbon source, nitrogen source, inorganic salts, growth factor) required by from bacterial classification target product fermenting and producing, precursor, defoamer, temperature, acid-base value, dissolved oxygen, inoculum concentration regulate and control to fermentation physical parameter, including shearing force (speed of agitator), feed-batch culture (carbon source, nitrogen source or precursor), two benches culture etc..
Metabolic activity in cells process exception is complicated, and this process is also that hundreds and thousands of kinds of enzymes steady ordered under interior exogenous signals intervention is adjusted reacting.Metal ion is often played an important role in this process, is prothetic group or the activator of enzyme.Some metal ions are the important component part in zymoprotein activated centre or necessary component；Some enzymes need metal ion to activate；Some transiting state metals transmit electronics by the change of itself chemical valence, participate in internal oxidation-reduction process.Therefore, add metal ion during fungi fermentation to be possible to affect the metabolism state of cell, and then be conducive to target product metabolism to synthesize.Existing document report, by adding some metal ions or slaine, is adjusted to the route of synthesis of secondary fungus metabolite, so that its fermentation yield improves.Such as, Jia etc. add magnesium ion improve cancer therapy drug ansamitocin P-3 yield (Bioresource Technology, 102 (21):10147-10150,2011)；The research such as Xu finds, calcium ion and sodium ion can effectively improve Ganoderma Lucidum metabolism synthesis ganoderic acid (Biotechnology Advances, 30 (6):1301-1308,2011；Biotechnology and Bioengineering 110(7):1913-1923,2013).
Content of the invention
The technical problem to be solved is to provide a kind of Jiangxi Cordyceps Militaris Cordyceps jiangxiensis JXPJ0109 bacterial strain and its application, and it enough effectively improves Jiangxi Cordyceps Militaris fermentating metabolism synthesis jiangxienone efficiency.
The present invention is realized in：Jiangxi Cordyceps Militaris Cordyceps jiangxiensis JXPJ0109 bacterial strain, its deposit number is CGMCC NO.9720.
The biology morphology feature of bacterial strain：This bacterium is in Czapek's medium (Sabouraud Dextrose Agar) (glucose 40g, peptone 10g, agar 18g, distilled water 1000mL, initial pH are 7) upper poor growth, cultivate 7 days at 25 DEG C, bacteria colony white, in irregular projection or wrinkle, quality is cotton-shaped, diameter 10mm about, have no conidium structure.Bacterium colony bottom reverse side is in reddish yellow, no soluble pigment.Colonial morphology is shown in accompanying drawing 1.
Using the method for Jiangxi Cordyceps Militaris Cordyceps jiangxiensis JXPJ0109 bacterial strain efficient metabolism synthesizing antitumor compound jiangxienone, comprise the steps：
1) by Jiangxi Cordyceps Militaris Cordyceps jiangxiensis JXPJ0109 inoculation in inclined-plane, cultivate 7-15 days at 24-28 DEG C, save backup at 4 DEG C；The composition of inclined-plane seed culture medium is calculated by mass percentage, including glucose 1-4%, wheat bran 0.1-1%, peptone 0.1-1%, dusty yeast 0.1-1%, potassium dihydrogen phosphate 0.05-0.15%, epsom salt 0.01-0.05% and agar 1.8%, total amount meets 100%, and initial pH is 7；
2) use transfer needle picking 0.5-1cm2Slant strains block is inoculated in shaking flask, is 24-28 DEG C in temperature, under conditions of rotating speed is 120-200rpm, shaking table culture 4-7 days；The liquid amount percent by volume of the liquid seed culture medium in shaking table is 30-60%, and the composition of liquid seed culture medium is calculated by mass percentage：Including glucose 1-4%, peptone 0.1-1%, dusty yeast 0.1-1%, epsom salt 0.01-0.05% and potassium dihydrogen phosphate 0.05-0.15%, dipotassium hydrogen phosphate 0.05-0.15%, total amount meets 100%, and initial pH is 7；
3) separately take a new shaking flask, fermentation medium is added in new shaking flask, liquid amount percent by volume is 40-70%, again by step 2) the middle seed culture fluid cultivating acquisition, it is inoculated in fermentation medium by the inoculum concentration for 3-5% for the percent by volume, is 120-200rpm in rotating speed, under conditions of temperature is 25-30 DEG C, lucifuge is cultivated 7-21 days, obtains finished product；Add in incubation that (whole process of culture can add ferrous ion through the degerming ferrous ion of 0.22 μm of filtering with microporous membrane, but the time of interpolation, the different effect that it promotes jiangxienone yield to increase was different, experiment proves that fermentation is added ferrous ion on the 8th day and promotes the effect that jiangxienone yield increases the most obvious, add during culture medium better than preparing), after interpolation, the concentration of ferrous ion is 0.1-100mM；The composition of fermentation medium is calculated by mass percentage, and including sucrose 1-4%, sodium nitrate 0.1-0.5%, dipotassium hydrogen phosphate 0.05-0.15%, epsom salt 0.01-0.05%, potassium chloride 0.01-0.05%, initial pH are 7.
Technique effect in order to verify the present invention is tested as follows：
First, mycelium dry weight, fermented sample are processed and jiangxienone content analysis method：
Mycelium dry weight：After fermentation terminates for 15 days, whole bottle cell culture fluid is carefully poured on the filter paper weighed in advance, is filtrated to get mycelium, clean at least 3 times with enough deionized waters, it is subsequently placed in 45 DEG C of baking ovens, dries to constant weight, weigh and calculate mycelium dry weight (DW/Biomass)；During sampling, respectively take three Duplicate Samples every time under same condition of culture, final result is with " mean value ± standard deviation "Represent.
Fermented sample processing method：Take the fresh mycelia cell after fermenting 15 days, pump large quantity of moisture with Buchner funnel, be placed in 45 DEG C of drying in oven, after about 24h, reach constant weight.The mycelial cell mortar dried grinds, and crosses 60 mesh sieves afterwards, uses polybag sealed packaging, 4 DEG C of preservations are standby.Precision weighs fermentation gained Jiangxi cordyceps mycelia dry powder 0.2g (accurately to 1mg), it is placed in 10mL centrifuge tube, water-saturated n-butanol is added to mix, ultrasonic extraction 60min, controls not higher than 45 DEG C of water temperature, ultrasonic extraction 2 times, after the completion of extraction, supernatant is placed in 45 DEG C of vacuum drying chambers to dry, then dried residue hplc grade methanol is dissolved, through 0.22 μm of organic filtering with microporous membrane, and use acetonitrile：Water (1:1, v/v) dilute, for high performance liquid chromatography quantitative analysis jiangxienone content.
Jiangxienone standard curve making：Precision weighs 5mg jiangxienone powder, is quantitatively dissolved in 10mL methyl alcohol, obtains the jiangxienone standard liquid of 0.5mg/mL, then with this standard liquid as mother liquor, is diluted to the series concentration such as 6.25,12.5,25,50,100,200g/mL further.Peak area according to each standard liquid of analysis method record of above-mentioned jiangxienone.With jiangxienone concentration as ordinate, peak area is abscissa, draws calibration curve, and obtains standard straight-line equation and R2.Calibration curve is shown in accompanying drawing 2.
Jiangxienone content analysis：The content of jiangxienone is analyzed using high performance liquid chromatography (HPLC) instrument.HPLC system uses Agilent 1200 type efficient liquid phase chromatographic analysis instrument, is equipped with light diode array (DAD) detector and chemoffice chromatographic work station, is also equipped with a quaternary pump, on-line degassing machine and hand sampling valve simultaneously；Chromatographic column adopts Shim pack PREP-ODS (II) ODS25 μm of RP18 (4.6 × 250mm, 5 μm of particle diameter).Analysis condition is：Flow velocity 1.0mL/min；40 DEG C of column temperature；Ultraviolet detection wavelength 219nm；Mobile phase is the gradient solution of water and methyl alcohol, and concrete gradient sees attached list 1.
Table 1 is efficient liquid phase gradient elution program
2nd, different metal ions add the impact to Jiangxi Chinese caterpillar fungus bacteria growing and jiangxienone yield：
In order to filter out optimal metal ion derivant, Jiangxi Cordyceps Militaris liquid deep layer fermenting initiates, and adds six kinds of conventional metal ions respectively：Sodium ion (sodium chloride), calcium ion (calcium chloride), ferrous ion (ferrous sulfate), manganese ion (manganese chloride), zinc ion (zinc chloride), magnesium ion (magnesium chloride), and investigate its impact to Jiangxi Chinese caterpillar fungus bacteria growing and jiangxienone metabolism synthesis.After the sterilizing 20min cooling of 121 DEG C of Preliminary fermentation culture medium, by metal ion through 0.22 μm of filtering with microporous membrane degerming after add stand-by.Each metal ion adds concentration：Sodium ion, calcium ion, manganese ion and magnesium ion add 1mM, 10mM, 100mM respectively；Zinc ion adds 1mM and 10mM；Ferrous ion adds 1.6mM and 4.8mM.Fermentation culture method such as embodiment, lucifuge measures the mycelium dry weight of fermented sample and the yield of jiangxienone after cultivating 15 days.Three Duplicate Samples are respectively taken every time, final result is with " mean value ± standard deviation " under same condition of cultureRepresent.
Shown in result such as accompanying drawing 3 (A) and 3 (B), 4.8mM ferrous ion raising jiangxienone yield is the most obvious, and its value is 221.26 ± 6.79 μ g/g；Meanwhile, mycelium dry weight is almost unchanged.100mM magnesium ion has certain facilitation to the growth of Jiangxi Cordyceps Militaris hyphal cell, but significantly reduces the yield of jiangxienone.Other metal ions are inconspicuous to Jiangxi Cordyceps Militaris growth effect, but the raising to jiangxienone yield is substantially lower than 4.8mM ferrous ion group.
Above-mentioned description of test, in six metal ion species of interpolation, ferrous ion can remarkably promote the metabolism synthesis of jiangxienone, improve its yield, and do not affect the growth of Jiangxi Cordyceps Militaris.3rd, ferrous ion adds the impact to jiangxienone yield for the time：
Investigate 4.8mM (final concentration) ferrous ion different time points and add the impact to jiangxienone yield.Ferrous ion through 0.22 μm of filtering with microporous membrane degerming after add fermentation medium, adding time point is the 0th day (fermentation is initial), 5 days (exponential phase) and 8 days (late exponential phase) respectively；In addition, adding within the 0th day equal-volume sterilized water as comparison.Fermentation culture method such as embodiment 1, lucifuge measures the yield of fermented sample jiangxienone after cultivating 15 days.Three Duplicate Samples are respectively taken every time, final result is with " mean value ± standard deviation " under same condition of cultureRepresent.
As shown in Figure 4, the 8th day (late exponential phase) adds ferrous ion and promotes the effect that jiangxienone metabolism synthesizes best result, and its yield increased nearly 2.9 times, and significant difference compared with control group.Adding within 0th day and the 5th day ferrous ion also has facilitation to jiangxienone metabolism synthesis, and its yield improves about 1.78～2.48 times compared with the control.Above example illustrates, three time points add ferrous ion and the metabolism of jiangxienone all can be promoted to synthesize, and interpolation facilitation effect is the most obvious within the 8th day, its yield highest.
4th, ferrous ion adds the impact to jiangxienone yield for the concentration：
Ferrous ion adds after fermentation through 0.22 μm of filtering with microporous membrane is degerming on the 8th day, and adding concentration is 3.2mM, 4.0mM, 4.8mM, 5.6mM and 6.4mM respectively, and adds equal-volume sterilized water as comparison.Fermentation culture method such as embodiment 1, lucifuge measures the yield of fermented sample jiangxienone after cultivating 15 days.Three Duplicate Samples are respectively taken every time, final result is with " mean value ± standard deviation " under same condition of cultureRepresent.
Result is as shown in Figure 5, except 8.0mM interpolation group, remaining ferrous ion interpolation group all improves the yield of jiangxienone, and wherein action effect is the most significantly 4.8mM interpolation group, its jiangxienone yield has reached 376.92 ± 15.33g/L, improves about 2.4 times compared with control group；And 1.6mM with 3.2mM interpolation group has been respectively increased 2.23 and 1.83 times compared with control group.Above example illustrates, the optimal interpolation concentration of ferrous ion is 4.8mM.During can determine that Jiangxi Cordyceps Militaris liquid deep layer fermenting produces jiangxienone by above example, the optimum adding conditional of ferrous ion：The interpolation time is fermented and cultured the 8th day, and interpolation concentration is 4.8mM.Under the conditions of being somebody's turn to do, the yield of jiangxienone is 376.92 ± 15.33 μ g/L, improves about 2.4 times compared with control group.
The Jiangxi Cordyceps Militaris Cordyceps jiangxiensis JXPJ0109 bacterial strain of the present invention has been deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC, address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3；Postcode:100101), its deposit number is CGMCC NO.9720, and preservation date is on October 14th, 2014, and Classification And Nomenclature is Jiangxi Chinese caterpillar fungus, and Latin name is Cordyceps jiangxiensis.
Present invention discover that Jiangxi Cordyceps Militaris Cordyceps jiangxiensis JXPJ0109 bacterial strain has good result to synthesis antitumoral compounds jiangxienone, and regulate and control this Cordyceps Militaris high-efficiency fermenting metabolism synthesizing antitumor compound jiangxienone by adding ferrous ion, such mode can effectively improve the efficiency that this Jiangxi Cordyceps Militaris fermentating metabolism synthesizes jiangxienone, yield brings up to the dry mycelium of 221.26 ± 6.79g/g by the dry mycelium of 1.0 μ g/g, greatly improves its yield.And the method, simply it is easy to operation, extremely suitable Cordyceps Militaris liquid deep layer fermenting production jiangxienone, to large-scale production jiangxienone and its patent medicine Journal of Sex Research, all has important theory significance and using value.The present invention has material source extensively, with low cost, the advantages of effect is preferable.
Brief description
Accompanying drawing 1 is colonial morphology on sabouraud culture medium for the Jiangxi Cordyceps Militaris
Accompanying drawing 2 is the calibration curve of jiangxienone sterling；
Accompanying drawing 3 adds the impact to Jiangxi Chinese caterpillar fungus bacteria growing and jiangxienone yield for different metal ions；
A：The impact to Jiangxi Chinese caterpillar fungus bacteria growing for the different metal ions；B：The impact to jiangxienone yield for the different metal ions；*P<0.05,**P<0.01vs control group；
Accompanying drawing 4 adds the impact to jiangxienone yield for the time for ferrous ion；
*P<0.05vs control group；
Accompanying drawing 5 adds the impact to jiangxienone yield for the concentration for ferrous ion；
*P<0.05vs control group.
Specific embodiment
Embodiments of the invention：The method promoting Cordyceps Militaris efficient metabolism synthesizing antitumor compound jiangxienone, comprises the steps：
1) Jiangxi Cordyceps Militaris are taken, bacterial classification is Jiangxi Cordyceps Militaris Cordyceps jiangxiensis JXPJ0109 bacterial strain, its deposit number is CGMCC No.9720, and bacterial classification long term storage is with 20% glycerol tube (bacterial culture fluid：Glycerine=8.0/2.0v/v), it is placed in low-temperature preservation in -74 DEG C of refrigerators；The preparation of inclined-plane seed culture medium：Glucose 30g, peptone 5g, dusty yeast 5g, potassium dihydrogen phosphate 1g, epsom salt 0.5g, wheat bran 5g and agar 18g, are settled to 1L, initial pH 7.0；Wheat bran is processed：By 5g wheat bran, add deionized water 0.5L, boil 30min with electric cooker, with eight layers of filtered through gauze, take filtrate；
Jiangxi Cordyceps Militaris are inoculated in inclined-plane, cultivate 7 days at 25 DEG C, save backup in the refrigerator being placed in 4 DEG C；
2) preparation of liquid seed culture medium：Glucose 30g, peptone 5g, dusty yeast 5g, epsom salt 0.5g and potassium dihydrogen phosphate 1g, dipotassium hydrogen phosphate 1g, it is settled to 1L (adjusting pH 7 before sterilizing)：Add the new liquid seed culture medium prepared of 100mL in the shaking flask of 250ml, with transfer needle picking 1cm2Slant strains block is inoculated in shaking flask, is 25 DEG C in temperature, under conditions of rotating speed is 150rpm, shaking table culture 5 days；
3)；The preparation of fermentation medium：Sucrose 30g, sodium nitrate 3g, dipotassium hydrogen phosphate 1g, epsom salt 0.5g and potassium chloride 0.5g, are settled to 1L (adjusting pH 7 before sterilizing)；Separately take the new shaking flask of a 250ml, 96ml fermentation medium is added in new shaking flask, again by step 2) the middle seed culture fluid cultivating acquisition, 4ml is in fermentation medium for inoculation, it is 160rpm in rotating speed, under conditions of temperature is 25 DEG C, lucifuge is cultivated 15 days, add the ferrous ion through 0.22 μm of sterilised membrane filter filtration sterilization when culture was to the 8th day, after interpolation, the concentration that adds of ferrous ion is 10mM.
Above-mentioned all cultures are standby based on sterilizing 20min at 121 DEG C.
Claims (2)
1. a kind of Jiangxi Cordyceps MilitarisCordyceps jiangxiensis
JXPJ0109 bacterial strain, its deposit number is CGMCC NO.9720.
2. a kind of method using bacterial strain as claimed in claim 1 efficient metabolism synthesizing antitumor compound jiangxienone it is characterised in that：Comprise the steps：
1）By Jiangxi Cordyceps MilitarisCordyceps jiangxiensis
JXPJ0109 inoculation, in inclined-plane, is cultivated 7-15 days, 4 at 24-28 DEG C
Save backup at DEG C；The composition of inclined-plane seed culture medium is calculated by mass percentage, in every kilogram of distilled water, 1-4% containing glucose, wheat bran 0.1-1%, peptone 0.1-1%, dusty yeast 0.1-1%, potassium dihydrogen phosphate 0.05-0.15%, epsom salt 0.01-0.05% and agar 1.8%, initial pH is 7；
2）With transfer needle picking 0.5-1 cm2Slant strains block is inoculated in shaking flask, is 24-28 DEG C in temperature, under conditions of rotating speed is 120-200 rpm, shaking table culture 4-7 days；The liquid amount percent by volume of the liquid seed culture medium in shaking table is 30-60%, the composition of liquid seed culture medium is calculated by mass percentage, in every kilogram of distilled water, 1-4% containing glucose, peptone 0.1-1%, dusty yeast 0.1-1%, epsom salt 0.01-0.05% and potassium dihydrogen phosphate 0.05-0.15%, dipotassium hydrogen phosphate 0.05-0.15%, initial pH are 7；
3）Separately take a new shaking flask, new shaking flask adds fermentation medium, liquid amount percent by volume is 40-70%, then by step 2）The seed culture fluid that middle culture obtains, is inoculated in fermentation medium by the inoculum concentration for 3-5% for the percent by volume, is 120-200rpm in rotating speed, under conditions of temperature is 25-30 DEG C, obtains finished product；Add in incubation through 0.22
The degerming ferrous ion of filtering with microporous membrane, after interpolation, the concentration of ferrous ion is 0.1-100 mM, and lucifuge is cultivated 7-21 days；The composition of fermentation medium is calculated by mass percentage, and including sucrose 1-4%, sodium nitrate 0.1-0.5%, dipotassium hydrogen phosphate 0.05-0.15%, epsom salt 0.01-0.05%, potassium chloride 0.01-0.05%, initial pH are 7.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102234232A (en) * | 2010-04-29 | 2011-11-09 | 遵义医学院附属医院 | Jiangxi ketene ester and application thereof |
| CN105821085A (en) * | 2015-01-23 | 2016-08-03 | 遵义医学院 | Method for promoting high-efficient metabolic synthesis of anti-neoplastic compound jiangxienone by cordyceps jiangxiensis |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102234232A (en) * | 2010-04-29 | 2011-11-09 | 遵义医学院附属医院 | Jiangxi ketene ester and application thereof |
| CN105821085A (en) * | 2015-01-23 | 2016-08-03 | 遵义医学院 | Method for promoting high-efficient metabolic synthesis of anti-neoplastic compound jiangxienone by cordyceps jiangxiensis |
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| Title |
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| 刘元元 等: "江西烯酮酯的发酵条件优化及定性定量检测", 《中国菌物学会第六届会员代表大会（2014年学术年会）暨贵州省食用菌产业发展高峰论坛会议摘要》 * |
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