CN105543125B - One plant of jute streptomycete for producing jinggangmeisu and its application - Google Patents
One plant of jute streptomycete for producing jinggangmeisu and its application Download PDFInfo
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Abstract
The invention discloses jute streptomycete (Streptomyces corchorusii) KN-1625 that one plant produces jinggangmeisu, are deposited in China typical culture collection center, and deposit number is CCTCC NO:M2015710.The bacterium can improve the content of jinggangmycin A in jinggangmeisu fermentation liquid, reduce the content of jinggangmeisu B, compared with starting strain, jinggangmycin A content, which improves 47%, B/A, reduces 37.56%.And jinggangmeisu stable yield, more than 30000 μ g/mL, the genetic stability of KN-1625 bacterial strain is good, and it is suitable with the first generation to reach four generations production jinggangmeisu level.
Description
Technical field
The invention belongs to field of biotechnology, and in particular to the jute streptomycete of one plant of production jinggangmeisu
(Streptomyces corchorusii), the invention further relates to the applications of the bacterial strain.
Background technique
Jinggangmeisu is a kind of farm antibiotics, can effectively prevent banded sclerotial blight, is " preventive effect is high, without phytotoxicity, pollution-free "
Eco-friendly pesticide is current yield and the maximum farm antibiotics of usable floor area.Jinggangmeisu belongs to aminoglycoside, mainly by
A, the groups such as B, C, D, E, F are grouped as, and find the compositions such as G and H again later, but only component A is most strong to the preventive and therapeutic effect of banded sclerotial blight,
High-content jinggangmycin A acts not only as Pesticide use, is also used as the intermediate of production medicine, application prospect is very
It is wide.However the jinggangmeisu impurity content at present using microbial fermentation processes production is higher, especially jinggangmeisu B's contains
Amount is higher, and product quality is generally poor.
Summary of the invention
The purpose of the present invention is by jute streptomyces strain carry out induced mutations, thus obtain one plant can be used not only for giving birth to
Jinggangmeisu is produced, and jinggangmycin A content can be improved, reduces the jute streptomycete of jinggangmeisu B content.
Applicant for starting strain, is used 1000Gy dosage 60CO- gamma-rays to carry out with jute streptomycete (KN-11)
First round mutagenesis, obtains one plant of KN-16 bacterial strain, which is higher by than the potency (content for producing jinggangmycin A) of starting strain
21%, the second wheel mutagenesis then is carried out to KN-16 bacterium with ultraviolet light again, obtains one plant of KN-162 bacterial strain, the potency than KN-16 mentions
It is high by 9.8%.KN-162 bacterial strain is subjected to resistance screening again, it should with the low own metabolism object processing of high-purity, impurity content
Bacterial strain kills or resists the breeding of many cells big absolutely, selects the anti feedback mutant strain for largely generating the metabolin, obtain 1 plant
Direct mutation KN-1625 bacterial strain, the content ratio KN-162 bacterial strain which produces jinggangmycin A is high by 18%, produces containing for jinggangmeisu B/A
Amount lower than KN-162 bacterial strain about 37.56%.
The bacterial strain is identified as jute streptomycete (Streptomyces corchorusii), is named as KN-1625, application
The bacterium is preserved in the China typical culture collection in the Wuhan University of Wuhan City, Hubei Province on November 26th, 2015 by people
Center (CCTCC), deposit number are CCTCC NO:M 2015710.
The performance of jinggangmeisu is produced for verifying KN-1625 bacterium, KN-1625 bacterial strain is put into production and carried out by applicant
The pilot scale research of two batches first activates KN-1625 bacterial strain, and the bacteria suspension after activation carries out secondary seed culture, training
Nutrient solution enters back into fermentor and ferments, and is detected with high performance liquid chromatography to fermentation liquid after putting tank, the results showed that:Fermentation
The content of jinggangmycin A is up to 3.065% in liquid, and the content ratio (B/A) of jinggangmeisu B and jinggangmycin A is only 11.52%,
Compared with starting strain (KN-11), jinggangmycin A content, which improves 47.00%, B/A, reduces 37.56%, and jinggangmeisu
Stable yield also increases more than 30000 μ g/mL, than CK bacterial strain.Test result is also shown that the heredity of KN1625 bacterial strain is steady
It is qualitative good, it is suitable with the first generation to reach four generations production jinggangmeisu level.
Specific embodiment
The present invention is described in detail below in conjunction with specific embodiment.
The mutagenesis and screening of 1 bacterial strain of embodiment
1. using 60CO- gamma-rays processing bacteria suspension to obtain mutant strain
The inclined-plane cell or appropriate spore for taking jute streptomycete KN-11, are placed in the sterile water containing bead, keep it thin
Born of the same parents or spore concentration sufficiently shake up and break up in 106~108/milliliter.Since γ firing line penetration capacity is very strong, cell is caused
Dead effect is more stronger than general chemical mutagen.Induced dosage, which rests in lethality 90-99%, to be advisable, and is to set out with KN-16
Bacterial strain, by its spore bacteria suspension various dose 60CO- gamma-ray irradiation, dosage 300Gy, 500Gy, 700Gy, 1000Gy,
When 1300Gy, 1500Gy, dead rate of making a speech respectively is 75.7%, 85.3%, 89.1%, 95.0%, 98.2%, 99.10%.
What processing spore death rate rose in 300Gy is very fast, and it is relatively slow that 500-1500Gy handles the variation of spore death rate.
Induced dosage is different, also different to the mutation rate of KN-11.Induced dosage is excessive, and direct mutation is very low, by 700Gy,
Bacteria suspension dilution after 1000Gy, 1300Gy, 1500Gy mutagenesis carries out plate count, is added 2 milliliters in the bacterial plaque after mutagenesis
Sterile water is impregnated 0.5 hour, purges bacterial plaque repeatedly with 1 milliliter of pipettor and be transferred in sterile test tube afterwards for several times, managed with this and be
10-1 pipe, takes turns doing 10 times of dilution spreads.28 DEG C constant temperature incubation 5 days, observe growing way, choosing colony diameter is big, growing way is fast, water suction
Fast, the full intermediate projections of lawn bacterium colonies carry out shaking flask screening, and one plant of KN-16 bacterial strain is obtained in 1000Gy Induced dosage,
For yield compared with starting strain (KN-11), potency is higher by 21%.
2. using ultraviolet mutagenesis processing screening mutant strain, to obtain the bacterial strain of a small number of function admirables
Bacterial strain after 60CO- gamma-ray and mutagenesis, most cells are dead, but there have the DNA in a few cell to occur to be prominent
Become, the bacterial strain after these are mutated carries out ultraviolet mutagenesis again, can influence the repair of DNA damage, is allowed to occur more
Mistake, improve induced mutation rate, mutagenic frequency is high and is not easy to reply, and ultraviolet light can cause the DNA mutation of microbial cells, is formed
Pyrimidine dimer.Illumination will disconnect dimer, so ultraviolet light carries out under the conditions of being protected from light when luring.Have previously been thought that lethality exists
90-99.9% effect is good.But current report thinks that lethality is conducive to the generation of direct mutation type in 70-80%, and screens
Bacterial strain genetic stability it is good.
10 × 108/ml monospore suspension is made in KN-16 bacterium, is taken in the aseptic flat board that 2ml is 90mm to diameter,
Ultraviolet mutagenesis (wavelength 253.7nm, power 15W, irradiation distance 30cm) is carried out under magnetic agitation, irradiation time is respectively 0,2,
5,8,10,15,20,25min, then take 28 DEG C of constant temperature on the bacteria suspension 0.1mL to plating medium by various dose irradiation
Culture 5 days, carries out count plate, and draw destruction curve after bacterium colony is grown, after obtaining destruction curve, is existed with lethality
The irradiation time of 80-90% irradiates bacterial strain, again 28 DEG C constant temperature incubation 5 days.Lethality is diluted and applied in the bacterial strain of 90-95%
On cloth to the bacterial strain containing maximum tolerance jinggangmycin A concentration, 28 DEG C constant temperature incubation 5 days, by the bacterium colony number shaking flask grown survey
Determine the content of jinggangmycin A and the content of B component, obtain one plant of KN-162 bacterial strain, shaking flask ratio KN-16 bacterial strain produces jinggangmycin A
Content is high by 9.8%.
3. obtaining the low bacterial strain of high yield jinggangmycin A, B component by resistance screening
Jinggangmycin A is aminoglycoside antibiotics, is the secondary metabolite of actinomyces, and metabolic pathway is complicated, grinds
Study carefully show antibiotic fermentation by feedback inhibition clearly, especially aminoglycoside antibiotics.Secondary metabolite is certain
One kind that biology generates in order to avoid detrimental effect caused by intermediate accumulations certain during primary metabolite is advantageous
In the metabolic type of existence.Since cometabolism and primary metabolite have common point poor intermediate, primary metabolite can be both synthesized
Product, and secondary metabolite can be synthesized.When nutriment is unbalanced in the environment of thalli growth, i.e., certain nutrition
Surplus, certain nutritional deficiency, thallus cannot effectively take in nutrition, under the action of metabolic system, the decline of growth and breeding rate,
And superfluous nutriment is transformed by the secondary metabolite unrelated with growth and breeding by the change of metabolic pathway.Therefore,
In bacterium, we just to eliminate feedback regulation of the metabolite to those common intermediates, are allowed to largely accumulate,
It is advantageous for the synthesis of jinggangmycin A in this way, reduces impurity content.
KN-162 bacterial strain is subjected to resistance screening again, is low itself of 18% impurity content with 64A high-purity, B/A
Metabolin jinggangmycin A (standard items) is put into growth training by various concentration (effective content 0.2,0.4,0.6,0.8,1.0,1.2)
Support and co-cultured in base, to kill or resist the breeding of many cells big absolutely, select largely generate the metabolin anti feedback it is prominent
Mutant.0.1ml is taken to be coated on the anti-plate of 4.0A after bacterium solution is diluted suitable concentration, 37 DEG C are cultivated 5-6 days, and picking 100 prominent
Mutant expands on normal inclined-plane, screens by shake flask fermentation, obtains 1 plant of positive mutating strain KN-1625 bacterial strain, which produces Jing Gang
The content ratio KN-162 bacterial strain of mycin A is high by 18%, produces the content ratio KN-162 bacterial strain low about 37.56% of jinggangmeisu B.
4. the identification of bacterial strain
The bacterial strain starting strain be jute streptomycete, gram-positive bacteria, after complex mutation, the strain morphology spore
Silk spiral shape, spore oval, surface is smooth, and bacterium colony is Slate grey, and pigment penetrates in culture medium, and base silk is dark brown, gas silk
Slate grey.Form is identical as starting strain, does not occur the change of Pseudomonas, and bacterial strain is identified still as jute streptomycete.
5. the genetic stability of bacterial strain
Improvement strain KN-1625 is connect into inclined-plane and carries out passage assays, to examine the genetic stability of improved strain, through passing 4
Show that the strain stability is good for confirmatory experiment.
The mitotic stability of 1 KN-1625 bacterial strain of table is examined
Data show that it is suitable with the first generation that KN-1625 bacterial strain reaches four generations production jinggangmeisu level, it was demonstrated that bacterial strain heredity
Stablize, can be used as engineering bacteria and put into production.
Embodiment 2 produces the experimental study of jinggangmeisu using bacterial strain
1) bacterial strain activates
The KN-1625 bacterial strain of preservation is activated on Gause I slant medium, 28 DEG C are cultivated 7 days, will with sterile water
Lawn is washed down, and seed bacteria suspension is obtained;
2) secondary seed culture
Seed bacteria suspension obtained by step 1) is accessed into seed fermentation tank, 200rpm, 40 DEG C of culture 16-18hr, seed fermentation
Medium component is:Rice meal 4.66%, corn flour 2.52%, dregs of beans 1.033%, NaCl 0.03%, CaCO3 0.05%,
KH2PO4 0.03%, surplus are water, and the content of each ingredient presses mass volume ratio;
3) fermented and cultured
The seed fermentation liquid obtained after step 2) fermentation is moved into bulk fermentation tank and is cultivated, setting fermentation parameter is:It is logical
Gas is than 1:1, mixing speed 420rpm, 40 DEG C of fermentation temperature, incubation time 30-32hr is obtained after putting tank containing jinggangmeisu
The ingredient of fermentation liquid, fermentation medium is:Rice meal 6.5%, starch 4%, corn flour 2.5%, dregs of beans 2.5%, NaCl
0.03%, CaCO3 0.03%, KH2PO4 0.03%, defoaming agent 0.01%, surplus are water, and the defoaming agent is XP-100F system
Column fermentation organic silicon defoamer, the content of each ingredient press mass volume ratio.
4) the HPLC detection of tank effective content and impurity content is put
HPLC testing conditions
Mobile phase:0.05mol/L disodium hydrogen phosphate-methanol=97 phosphoric acid (PH7.0):3
Chromatographic column:TC-C18/5.8 200X4.6mm
Detector:Ultraviolet 210nm
Flow velocity:1.0ml.min-1
Retention time:About 5.0min
By jinggangmycin A standard items be made into 25,50,75,100, the standard solution of five kinds of different contents of 125ug/ml,
It determines and is examined under chromatographic condition, be that ordinate draws working curve using jinggangmycin A amount as abscissa, peak area, obtain a line
The good straight line of sexual intercourse, equation of linear regression Y=7.6196X+36.675, suitable coefficients R 2=0.9995.Illustrate that well ridge is mould
Plain A concentration in 25~125ug/ml range, peak area and jinggangmycin A is dense cross between have good linear relationship.
5) data are analyzed
The standard sample spectrum of 3 kinds of concentration is recalled in chromatographic work station, carries out calibrated, drafting standard curve.According to jinggangmeisu
A, the content ratio of the calculated by peak area B/A of B, while according to the content of control and standard curve calculating jinggangmycin A, detect two batches
The data of secondary production, while being control with starting strain, as a result it see the table below.
Liquid phase peak area and the data statistics of 2 KN-1625 bacterial strain of table and control strain
It is shown by 2 data of table, jinggangmycin A content average out to 2.085% in the fermentation liquid of CK bacterial strain production, B/A is average
It is 18.45%;Jinggangmycin A content average out to 3.065 in the fermentation liquid of KN-1625 bacterial strain production, B/A average out to 11.52%;
The jinggangmycin A content ratio CK bacterial strain of KN-1625 bacterial strain, which improves 47.00%, B/A, reduces 37.56%.
The jinggangmeisu stable yield of KN-1625 bacterial strain production also increases more than 30000 μ g/mL than CK bacterial strain.
Claims (4)
1. jute streptomycete (Streptomyces corchorusii) KN-1625 of one plant of production jinggangmeisu, is deposited in China
Type Tissue Collection, deposit number are CCTCC NO:M 2015710.
2. application of the jute streptomycete KN-1625 described in claim 1 in jinggangmeisu production.
3. a kind of jinggangmeisu production method, it is characterised in that:It is hair with jute streptomycete KN-1625 described in claim 1
Yeast-like fungi strain.
4. jinggangmeisu production method as claimed in claim 3, it is characterised in that include the following steps:
1) bacterial strain activates:The jute streptomycete KN-1625 bacterial strain is activated on Gause I slant medium, 28 DEG C of cultures
7 days, lawn is washed down with sterile water, obtains seed bacteria suspension;
2) secondary seed culture:Seed bacteria suspension is accessed into seed fermentation tank, 200rpm, 40 DEG C of 16~18hr of culture, seed hair
Ferment medium component is:Rice meal 4.66%, corn flour 2.52%, dregs of beans 1.033%, NaCl 0.03%, CaCO3
0.05%, KH2PO40.03%, surplus is water, and the content of each ingredient presses mass volume ratio;
3) fermented and cultured:The seed fermentation liquid obtained after step 2) fermentation is moved into bulk fermentation tank and is cultivated, setting fermentation ginseng
Number is:Ventilation ratio 1:1, mixing speed 420rpm, 40 DEG C of fermentation temperature, 30~32hr of incubation time, the ingredient of fermentation medium
For:Rice meal 6.5%, starch 4%, corn flour 2.5%, dregs of beans 2.5%, NaCl 0.03%, CaCO30.03%, KH2PO4
0.03%, defoaming agent 0.01%, surplus is water, and the content of each ingredient presses mass volume ratio, is obtained after fermentation containing well
The fermentation liquid of ridge mycin.
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Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5662898A (en) * | 1990-08-20 | 1997-09-02 | Ciba-Geigy Corporation | Genes for the synthesis of antipathogenic substances |
CN102010835A (en) * | 2010-07-27 | 2011-04-13 | 江苏丘陵地区镇江农业科学研究所 | Streptomyces corchorusii strain NF0919, purpose and preparation method of active zymotic fluid thereof |
CN103937856A (en) * | 2014-04-21 | 2014-07-23 | 浙江大学 | Fermentation method capable of enhancing validamycin yield |
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- 2015-12-21 CN CN201510963611.0A patent/CN105543125B/en active Active
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5662898A (en) * | 1990-08-20 | 1997-09-02 | Ciba-Geigy Corporation | Genes for the synthesis of antipathogenic substances |
CN102010835A (en) * | 2010-07-27 | 2011-04-13 | 江苏丘陵地区镇江农业科学研究所 | Streptomyces corchorusii strain NF0919, purpose and preparation method of active zymotic fluid thereof |
CN103937856A (en) * | 2014-04-21 | 2014-07-23 | 浙江大学 | Fermentation method capable of enhancing validamycin yield |
Non-Patent Citations (2)
Title |
---|
Butalactin: a new butanolide antibiotic from streptomyces corchorusii;Sugata Chatterjee等;《Tetrahedron Letters》;19910101;第32卷(第1期);141-144 * |
黄麻链霉菌NF0919菌株发酵提取液HPLC 检测分析及稳定性研究;张文文 等;《中国农学通报》;20141231;第30卷(第33期);145-149 * |
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