CN103757069A - Method for producing prodigiosin by coupling resin adsorption in-situ separation with fermentation - Google Patents
Method for producing prodigiosin by coupling resin adsorption in-situ separation with fermentation Download PDFInfo
- Publication number
- CN103757069A CN103757069A CN201410010765.3A CN201410010765A CN103757069A CN 103757069 A CN103757069 A CN 103757069A CN 201410010765 A CN201410010765 A CN 201410010765A CN 103757069 A CN103757069 A CN 103757069A
- Authority
- CN
- China
- Prior art keywords
- prodigiosin
- adsorption
- fermentation
- fermented liquid
- resin
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a method for producing prodigiosin by coupling resin adsorption in-situ separation with fermentation. The method comprises the following steps: performing fermentation culture on prodigiosin through two-level fermentation, namely performing flat activation, seed culture and final amplification culture; performing a single factor experiment according to the sample loading quantity, pH of adsorption liquid, adsorption time and adsorption temperature, and measuring the adsorption rate; and performing an orthogonal experiment according to the experimental factor of the orthogonal experiment determined by the single factor experiment. According to the method disclosed by the invention, the screened high-yield prodigiosin strain is used as an original strain, a culture medium optimized by a research team is adopted for shake flask fermentation, and the optimal adsorption resin is screened; through the design of single factor and orthogonal experiment, the prodigiosin adsorption of the resin is observed, and the best adsorption conditions are determined to provide a basis for coupling the fermentation separation of prodigiosin.
Description
Technical field
The invention belongs to technological field of biochemistry, relate in particular to a kind of method that resin absorption original position is separated and coupled fermentation is produced prodigiosin.
Background technology
Nowadays very large to the Research Requirements of prodigiosin on market, it has good effect as the lipopolysaccharides extracting in natural biological to anticancer and tumour.And domestic market is mostly the haematochrome extracting from plant, to compare bacterium and produce, the plant extract cycle is long, and complex process can impact the quality of haematochrome and output.Although the Serratia in laboratory has higher output simultaneously.But obtaining sterling, separation must pass through resin chromatographic separation, although effect is better, consuming time of a specified duration, be unfavorable for large-scale industrialization production.
So be a solution preferably from common industrial fermentation separation coupling method as sought the method for sharp separation absorption method separation.Therefore, for the separation method of prodigiosin, need to constantly innovate, find optimum separation coupling route also very necessary.
Summary of the invention
The object of the present invention is to provide a kind of method that resin absorption original position is separated and coupled fermentation is produced prodigiosin, being intended to provides foundation for the Integrated process of prodigiosin.
The present invention is achieved in that a kind of resin absorption original position method separated and coupled fermentation production prodigiosin comprises the following steps:
Step 1, employing second order fermentation carry out the fermentation culture of prodigiosin: first carry out flat board activation, next carries out seed culture, last enlarged culturing;
Step 3, according to the experimental factor of the definite orthogonal test of single factor experiment, carry out orthogonal test.
Further, the laboratory apparatus that the separated also coupled fermentation of described resin absorption original position is produced the method for prodigiosin comprises: 722s visible spectrophotometer, electronic balance AL204, high speed centrifugation VS-15000N, HD-930 built-up type total temperature concussion incubator, pH meter UB-7.
The test reagent that the separated also coupled fermentation of described resin absorption original position is produced the method for prodigiosin comprises: D101 macroporous adsorbent resin, hydrochloric acid, acetone analytical pure, peptone, glycerine, NaCl, MgSO4, glycine etc.
Further, to carry out the concrete steps of fermentation culture of prodigiosin as follows for the employing second order fermentation described in step 1:
First dull and stereotyped activation: the bacterial classification in subzero 70 degree preservations, at ready flat lining out, is cultivated 12 hours for 37 ℃;
Secondly seed culture: select in single bacterium colony access shaking flask that growing way on flat board is good and cultivate, 37 ℃, 200r/min, 12 hours;
Last enlarged culturing: by the accurate shaking flask preparing of seed culture fluid access, 28 ℃, 200r/min, 48 hours.
Further, the concrete grammar that the adsorption rate described in step 2 is measured is as follows:
To test gained prodigiosin acetone extract rotary evaporation to constant weight, accurately take product 0.0400g, then with acetone, be settled in 50ml volumetric flask, make 0.8g/L storing solution, with liquid-transfering gun, accurately draw 150ul, 200ul, 250ul, 300ul, 350ul storing solution is in 10ml volumetric flask, with acetone constant volume, make 0.012g/L, 0.016g/L, 0.020g/L, 0.024g/L, 0.028g/L solution, under 535nm, survey its OD value, take OD value as X-coordinate, product concentration is ordinate zou curve plotting;
D101 to the adsorption equation of prodigiosin is
(1)
O is adsorption rate (g/g), and C1 is the content (g/L) of prodigiosin in fermented liquid, and C2 is the content (g/L) of prodigiosin in absorption secondary fermentation liquid, and M is the D101 quality (g) of putting into fermented liquid, and V is fermentating liquid volume (L).
Further, the concrete grammar of the single factor experiment described in step 2 is as follows:
Test for applied sample amount: accurately take six parts of about D101 macroporous adsorbent resin 0.5g that activated with ethanol, putting into respectively fermentating liquid volume is 10,20,30,40,50,60ml shaking flask, puts into 28 ℃, shakes approximately 20 hours in the constant-temperature table of 200r/min, dilute in the lump 100 times with the fermented liquid not adsorbing afterwards and measure OD value, according to formula (1), survey adsorption rate.
Test for pH: accurately take six parts of about D101 macroporous adsorbent resin 0.5g that activated with ethanol, put into respectively and contain 10ml fermented liquid pH and be respectively 1,2,3,4, in 5,6,7 shaking flask, put into 28 ℃, in the constant-temperature table of 200r/min, shake approximately 20 hours, dilute in the lump 100 times with the fermented liquid not adsorbing afterwards and measure OD value, according to formula (1), survey adsorption rate.
Test for adsorption time: accurately take six parts of about D101 macroporous adsorbent resin 0.5g that activated with ethanol, put into respectively the shaking flask that contains 10ml fermented liquid, put into 28 ℃, in the constant-temperature table of 200r/min, shake, earthquake 1 respectively, 5,10,15,20, after 25h, dilute 100 times and measure OD value, the fermented liquid not adsorbing dilutes in the lump 100 times and measures OD value, according to formula (1), surveys adsorption rate.
Test for adsorption temp: accurately take five parts of about D101 macroporous adsorbent resin 0.5g that activated with ethanol, put into respectively contain 10ml fermented liquid shaking flask 15,20,25,30,35 ℃, in shaking table, shake and dilute 100 times after 20h and measure OD value, the fermented liquid not adsorbing dilutes in the lump 100 times and measures OD value, according to formula (1), surveys adsorption rate.
effect gathers
The present invention take that to filter out high yield haematochrome bacterial strain be starting strain, the substratum that adopts research team to optimize carries out shake flask fermentation, screen optimum polymeric adsorbent, by single factor and orthogonal experimental design, carry out the investigation of resin to prodigiosin absorption property, and determine best adsorption conditions, for the Integrated process of prodigiosin provides foundation.
Accompanying drawing explanation
Fig. 1 is the method flow schema that the separated and coupled fermentation of the resin absorption original position that provides of the embodiment of the present invention is produced prodigiosin;
Fig. 2 is the influence curve of the applied sample amount that provides of the embodiment of the present invention to adsorption rate;
Fig. 3 is the influence curve of the adsorption time that provides of the embodiment of the present invention to prodigiosin adsorption rate;
Fig. 4 is the influence curve of the pH value that provides of the embodiment of the present invention to adsorption rate;
Fig. 5 is the influence curve of the temperature that provides of the embodiment of the present invention to adsorption rate.
Embodiment
In order to make object of the present invention, technical scheme and advantage clearer, below in conjunction with drawings and Examples, the present invention is further elaborated.Should be appreciated that specific embodiment described herein, only in order to explain the present invention, is not intended to limit the present invention.
Fig. 1 shows the method flow that the separated also coupled fermentation of a kind of resin absorption original position of the present invention is produced prodigiosin, comprises the following steps:
S101: adopt second order fermentation to carry out the fermentation culture of prodigiosin: first carry out flat board activation, next carries out seed culture, last enlarged culturing;
S102: carry out single factor experiment for applied sample amount, adsorption liquid pH, adsorption time, adsorption temp, and measure adsorption rate;
S103: carry out orthogonal test according to the experimental factor of the definite orthogonal test of single factor experiment.
Further, the laboratory apparatus that the separated also coupled fermentation of described resin absorption original position is produced the method for prodigiosin comprises: 722s visible spectrophotometer, electronic balance AL204, high speed centrifugation VS-15000N, HD-930 built-up type total temperature concussion incubator, pH meter UB-7.
The test reagent that the separated also coupled fermentation of described resin absorption original position is produced the method for prodigiosin comprises: D101 macroporous adsorbent resin, hydrochloric acid, acetone analytical pure, peptone, glycerine, NaCl, MgSO4, glycine etc.
Further, to carry out the concrete steps of fermentation culture of prodigiosin as follows for the employing second order fermentation described in step S101:
First dull and stereotyped activation: the bacterial classification in subzero 70 degree preservations, at ready flat lining out, is cultivated 12 hours for 37 ℃;
Secondly seed culture: select in single bacterium colony access shaking flask that growing way on flat board is good and cultivate, 37 ℃, 200r/min, 12 hours;
Last enlarged culturing: by the accurate shaking flask preparing of seed culture fluid access, 28 ℃, 200r/min, 48 hours.
Further, the concrete grammar that the adsorption rate described in step S102 is measured is as follows:
To test gained prodigiosin acetone extract rotary evaporation to constant weight, accurately take product 0.0400g, then with acetone, be settled in 50ml volumetric flask, make 0.8g/L storing solution, with liquid-transfering gun, accurately draw 150ul, 200ul, 250ul, 300ul, 350ul storing solution is in 10ml volumetric flask, with acetone constant volume, make 0.012g/L, 0.016g/L, 0.020g/L, 0.024g/L, 0.028g/L solution, under 535nm, survey its OD value, take OD value as X-coordinate, product concentration is ordinate zou curve plotting;
D101 to the adsorption equation of prodigiosin is
(1)
O is adsorption rate (g/g), and C1 is the content (g/L) of prodigiosin in fermented liquid, and C2 is the content (g/L) of prodigiosin in absorption secondary fermentation liquid, and M is the D101 quality (g) of putting into fermented liquid, and V is fermentating liquid volume (L).
Further, the concrete grammar of the single factor experiment described in step S102 is as follows:
Test for applied sample amount: accurately take six parts of about D101 macroporous adsorbent resin 0.5g that activated with ethanol, putting into respectively fermentating liquid volume is 10,20,30,40,50,60ml shaking flask, puts into 28 ℃, shakes approximately 20 hours in the constant-temperature table of 200r/min, dilute in the lump 100 times with the fermented liquid not adsorbing afterwards and measure OD value, according to formula (1), survey adsorption rate.
Test for pH: accurately take six parts of about D101 macroporous adsorbent resin 0.5g that activated with ethanol, put into respectively and contain 10ml fermented liquid pH and be respectively 1,2,3,4, in 5,6,7 shaking flask, put into 28 ℃, in the constant-temperature table of 200r/min, shake approximately 20 hours, dilute in the lump 100 times with the fermented liquid not adsorbing afterwards and measure OD value, according to formula (1), survey adsorption rate.
Test for adsorption time: accurately take six parts of about D101 macroporous adsorbent resin 0.5g that activated with ethanol, put into respectively the shaking flask that contains 10ml fermented liquid, put into 28 ℃, in the constant-temperature table of 200r/min, shake, earthquake 1 respectively, 5,10,15,20, after 25h, dilute 100 times and measure OD value, the fermented liquid not adsorbing dilutes in the lump 100 times and measures OD value, according to formula (1), surveys adsorption rate.
Test for adsorption temp: accurately take five parts of about D101 macroporous adsorbent resin 0.5g that activated with ethanol, put into respectively contain 10ml fermented liquid shaking flask 15,20,25,30,35 ℃, in shaking table, shake and dilute 100 times after 20h and measure OD value, the fermented liquid not adsorbing dilutes in the lump 100 times and measures OD value, according to formula (1), surveys adsorption rate.
Below in conjunction with drawings and the specific embodiments, the present invention will be further described.
As seen from Figure 3, along with the adsorption rate of the increase macroporous resin of applied sample amount is also rising, so applied sample amount while reaching 40ml ascensional range not obvious, reach 40,50,60ml applied sample amount adsorption rate and significantly do not increased, therefore select 40ml, be best applied sample amount.
Result in Fig. 4 can be found out in the lifting of front ten hours adsorption rates of absorption comparatively obvious, 15h, and 20h promotes not obvious to adsorption rate, may be that resin absorption approaches saturated, but still have increase adsorption rate.So select 20h to be adsorbed as the optimal adsorption time.
As seen from Figure 5 pH respectively 1 and the adsorption effect of 7 o'clock poor, and be 2,3,4 at pH, 5,6 o'clock adsorption rates approach and at pH 2 with there is high value at 3 o'clock, this may be acid relevant with the optimum range of prodigiosin, so selection pH=3 is optimal adsorption pH.
Analysis can obtain: when 15,20 ℃ of lesser tempss, adsorption rate is lower, and is 25,30 in temperature, although adsorption rate increases and changes not obviously 35 ℃ time, 25 ℃ is that adsorption rate is higher, therefore select 25 ℃, is optimal adsorption temperature.
After selected orthogonal table, by the level of factor table of four factor three levels, test, and result analyzed, obtain following table:
From orthogonal test, the time has the greatest impact to adsorption rate, is secondly pH and applied sample amount, and adsorption temp is minimum on the impact of adsorption rate.
Adsorption rate along with the increase D101 resin of applied sample amount is also rising as seen from Figure 2, but ascensional range obviously reduces after 30ml applied sample amount, afterwards 40,50,60ml applied sample amount adsorption rate has not significantly increased, therefore select 40ml, is best applied sample amount; Result in Fig. 3 can be found out in the lifting of front ten hours adsorption rates of absorption comparatively obvious, 15h, and 20h promotes not obvious to adsorption rate, may be that resin absorption approaches saturated, but still have increase adsorption rate.So select 20h to be adsorbed as the optimal adsorption time; As seen from Figure 4 pH respectively 1 and the adsorption effect of 7 o'clock poor, and be 2,3,4 at pH, 5,6 o'clock adsorption rates approach and at pH 2 with there is high value at 3 o'clock, this may be acid relevant with the optimum range of prodigiosin, so selection pH=3 is optimal adsorption pH; When 15,20 ℃ of lesser tempss, adsorption rate is lower as seen from Figure 5, and is 25,30 in temperature, although adsorption rate increases and changes not obviously 35 ℃ time, from figure, 25 ℃ is that adsorption rate is higher, therefore select 25 ℃, is optimal adsorption temperature.
Therefore when D101 macroporous adsorbent resin quality is 0.5g left and right, optimal adsorption condition is pH=2, fermented liquid 50ml, 30 ℃, adsorption time 25h, adsorption rate is the prodigiosin of every gram of D101 resin absorption 0.1455.
Although above-mentioned, by reference to the accompanying drawings the specific embodiment of the present invention is described; but be not limiting the scope of the invention; one of ordinary skill in the art should be understood that; on the basis of technical scheme of the present invention, those skilled in the art do not need to pay various modifications that performing creative labour can make or distortion still within protection scope of the present invention.
Claims (5)
1. the method that resin absorption original position is separated and coupled fermentation is produced prodigiosin, is characterized in that, described research method comprises the following steps:
Step 1, employing second order fermentation carry out the fermentation culture of prodigiosin: first carry out flat board activation, next carries out seed culture, last enlarged culturing;
Step 2, for applied sample amount, adsorption liquid pH, adsorption time, adsorption temp, carry out single factor experiment, and measure adsorption rate;
Step 3, according to the experimental factor of the definite orthogonal test of single factor experiment, carry out orthogonal test.
2. the method that resin absorption original position as claimed in claim 1 is separated and coupled fermentation is produced prodigiosin, it is characterized in that, the laboratory apparatus that the separated also coupled fermentation of described resin absorption original position is produced the method for prodigiosin comprises: 722s visible spectrophotometer, electronic balance AL204, high speed centrifugation VS-15000N, HD-930 built-up type total temperature concussion incubator, pH meter UB-7;
The test reagent that the separated also coupled fermentation of described resin absorption original position is produced the method for prodigiosin comprises: D101 macroporous adsorbent resin, hydrochloric acid, acetone analytical pure, peptone, glycerine, NaCl, MgSO4, glycine.
3. the method that resin absorption original position as claimed in claim 1 is separated and coupled fermentation is produced prodigiosin, is characterized in that, the concrete steps of fermentation culture that the employing second order fermentation described in step 1 carries out prodigiosin are as follows:
First dull and stereotyped activation: the bacterial classification in subzero 70 degree preservations, at ready flat lining out, is cultivated 12 hours for 37 ℃;
Secondly seed culture: select in single bacterium colony access shaking flask that growing way on flat board is good and cultivate, 37 ℃, 200r/min, 12 hours;
Last enlarged culturing: by the accurate shaking flask preparing of seed culture fluid access, 28 ℃, 200r/min, 48 hours.
4. the method that resin absorption original position as claimed in claim 1 is separated and coupled fermentation is produced prodigiosin, is characterized in that, the concrete grammar that the adsorption rate described in step 2 is measured is as follows:
Gained prodigiosin acetone extract rotary evaporation, to constant weight, is taken to product 0.0400g, then with acetone, be settled in 50ml volumetric flask, make 0.8g/L storing solution, with liquid-transfering gun, accurately draw 150ul, 200ul, 250ul, 300ul, 350ul storing solution is in 10ml volumetric flask, with acetone constant volume, make 0.012g/L, 0.016g/L, 0.020g/L, 0.024g/L, 0.028g/L solution, under 535nm, survey its OD value, take OD value as X-coordinate, product concentration is ordinate zou curve plotting;
D101 to the adsorption equation of prodigiosin is
O is adsorption rate, and unit is g/g, and C1 is the content of prodigiosin in fermented liquid, and unit is g/L, C2 is the content of prodigiosin in absorption secondary fermentation liquid, and unit is g/L, and M is the D101 quality of putting into fermented liquid, unit is g, and V is fermentating liquid volume, and unit is L.
5. the method that resin absorption original position as claimed in claim 1 is separated and coupled fermentation is produced prodigiosin, is characterized in that, the concrete grammar of the single factor experiment described in step 2 is as follows:
Test for applied sample amount: take six parts of the D101 macroporous adsorbent resin 0.5g that activated with ethanol, putting into respectively fermentating liquid volume is 10,20,30,40,50,60ml shaking flask, puts into 28 ℃, shakes 20 hours in the constant-temperature table of 200r/min, dilute in the lump 100 times with the fermented liquid not adsorbing afterwards and measure OD value, according to formula
survey adsorption rate;
Test for pH: take six parts of the D101 macroporous adsorbent resin 0.5g that activated with ethanol, put into respectively and contain 10ml fermented liquid pH and be respectively 1,2,3,4, in 5,6,7 shaking flask, put into 28 ℃, in the constant-temperature table of 200r/min, shake 20 hours, dilute in the lump 100 times with the fermented liquid not adsorbing afterwards and measure OD value, according to formula
survey adsorption rate;
Test for adsorption time: take six parts of the D101 macroporous adsorbent resin 0.5g that activated with ethanol, put into respectively the shaking flask that contains 10ml fermented liquid, put into 28 ℃, in the constant-temperature table of 200r/min, shake, earthquake 1 respectively, 5,10,15,20, after 25h, dilute 100 times and measure OD value, the fermented liquid not adsorbing dilutes in the lump 100 times and measures OD value, according to formula
Survey adsorption rate;
Test for adsorption temp: take five parts of the D101 macroporous adsorbent resin 0.5g that activated with ethanol, put into respectively contain 10ml fermented liquid shaking flask 15,20,25,30,35 ℃, in shaking table, shake and dilute 100 times after 20h and measure OD value, the fermented liquid not adsorbing dilutes in the lump 100 times and measures OD value, according to formula
survey adsorption rate.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410010765.3A CN103757069A (en) | 2014-01-09 | 2014-01-09 | Method for producing prodigiosin by coupling resin adsorption in-situ separation with fermentation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410010765.3A CN103757069A (en) | 2014-01-09 | 2014-01-09 | Method for producing prodigiosin by coupling resin adsorption in-situ separation with fermentation |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103757069A true CN103757069A (en) | 2014-04-30 |
Family
ID=50524385
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410010765.3A Pending CN103757069A (en) | 2014-01-09 | 2014-01-09 | Method for producing prodigiosin by coupling resin adsorption in-situ separation with fermentation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103757069A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104480224A (en) * | 2014-12-13 | 2015-04-01 | 河南天冠企业集团有限公司 | Method for regulating PH value in microbial fermentation process |
| CN104711295A (en) * | 2015-03-14 | 2015-06-17 | 国家海洋局第一海洋研究所 | Method for utilizing macro-porous adsorption resin to adsorb coloring generated from fungi through dynamic fermentation |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1563400A (en) * | 2004-04-19 | 2005-01-12 | 华东理工大学 | Method for separating and preparing prodigiosin |
| CN102002469A (en) * | 2010-09-28 | 2011-04-06 | 嘉兴学院 | Bacterial strain for producing prodigiosin and method thereof |
-
2014
- 2014-01-09 CN CN201410010765.3A patent/CN103757069A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1563400A (en) * | 2004-04-19 | 2005-01-12 | 华东理工大学 | Method for separating and preparing prodigiosin |
| CN102002469A (en) * | 2010-09-28 | 2011-04-06 | 嘉兴学院 | Bacterial strain for producing prodigiosin and method thereof |
Non-Patent Citations (4)
| Title |
|---|
| XUEDONG WANG ET AL.: "Development of an adsorption procedure for the direct separation and purification of prodigiosin from culture broth", 《BIOTECHNOL. APPL. BIOCHEM.》, vol. 40, no. 3, 31 December 2004 (2004-12-31) * |
| 何志伟等: "大孔吸附树脂纯化乌饭树果色素的研究", 《食品科技》, no. 9, 20 September 2007 (2007-09-20), pages 88 - 92 * |
| 王增等: "灵菌红素提取工艺的优化方案", 《嘉兴学院学报》, vol. 23, no. 6, 11 November 2011 (2011-11-11), pages 96 - 99 * |
| 黄永春等: "采用大孔吸附树脂D-101初步分离提取TJ430菌株发酵液中的抗生素", 《植物保护》, vol. 39, no. 4, 8 August 2013 (2013-08-08), pages 85 - 89 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104480224A (en) * | 2014-12-13 | 2015-04-01 | 河南天冠企业集团有限公司 | Method for regulating PH value in microbial fermentation process |
| CN104711295A (en) * | 2015-03-14 | 2015-06-17 | 国家海洋局第一海洋研究所 | Method for utilizing macro-porous adsorption resin to adsorb coloring generated from fungi through dynamic fermentation |
| CN104711295B (en) * | 2015-03-14 | 2018-05-25 | 国家海洋局第一海洋研究所 | A kind of method of macroporous absorbent resin dynamic fermentation absorption fungi pigment |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102084780B (en) | Method for culturing cordyceps militaris sporocarp with high-content bioactive substances | |
| CN104178564B (en) | The screening technique of brown paddy plant hopper reference gene and application under high temperature stress | |
| CN101671712B (en) | Method and device for optimizing and amplifying abamectin fermenting process | |
| CN103757069A (en) | Method for producing prodigiosin by coupling resin adsorption in-situ separation with fermentation | |
| CN204028066U (en) | A kind of normal pressure of soil release gas cultivates analogue means | |
| CN111134007B (en) | Method for improving flavone content in adventitious roots of northeast thorn ginseng | |
| CN101398413B (en) | Liquid phase analytical method for iminodiacetic acid | |
| CN103674929B (en) | Spectrum analysis is for the examination method of plant seedlings | |
| CN102937631A (en) | Method for screening active ingredients of traditional Chinese medicine by nano mesoporous material immobilized target protein | |
| Sun et al. | Analysis of metabolomic changes in xylem and phloem sap of cucumber under phosphorus stresses | |
| CN107629980A (en) | The Ludwig enterobacteria SXZ N5 bacterial strains and purposes of one plant of production huperzine and Huperzine B | |
| CN102417888B (en) | Clostridium acetobutylicum for producing butanol by utilizing manihot as raw materials and application thereof | |
| CN103911407A (en) | Marine fungus-derived Azaphilone dimer compound, and preparation method and application thereof | |
| CN105628631A (en) | Rapid detection method of biological catalytic conversion rate of hydrocortisone | |
| CN109060995A (en) | A kind of screening technique of almond perfume (or spice) Tea Germplasm | |
| CN103323551A (en) | Method for detecting content of medlar acid | |
| CN107523522A (en) | The bacillus amyloliquefaciens SXZ N2 bacterial strains and purposes of one plant of production huperzine and Huperzine B | |
| CN103642691B (en) | Deep-hole cell culture plate | |
| CN103509737B (en) | Morganella morganii and application thereof to fermentation detoxification of barbadosnut cake | |
| CN101974435A (en) | Neomycin strain breeding and rapid screening method | |
| CN104569185B (en) | A kind of utilize the method for citric acid in ion exclusion chromatography's post detection edible fungi culture fluid | |
| CN104569182B (en) | A kind of method utilizing ion exclusion chromatography's post detection edible fungi culture fluid mesotartaric acid | |
| CN104569183B (en) | A kind of method utilizing ion exclusion chromatography's post detection edible fungi culture fluid mesoxalic acid | |
| CN104350949A (en) | Cordyceps sinensis liquid-state fermentation process | |
| Marques et al. | Assessment of the potential of sunflower grown in metal-contaminated soils for production of biofuels |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20140430 |