CN102586134A - Marine streptomyces viridochromogenes strain for producing alkali-tolerant and salt-tolerant xylanase and application of marine streptomyces viridochromogenes strain - Google Patents
Marine streptomyces viridochromogenes strain for producing alkali-tolerant and salt-tolerant xylanase and application of marine streptomyces viridochromogenes strain Download PDFInfo
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Abstract
A marine streptomyces viridochromogenes strain for producing alkali-tolerant and salt-tolerant xylanase and application of the marine streptomyces viridochromogenes strain belong to the field of biotechnology. The strain is collected in the China General Microbiological Culture Collection Center, and the collection number is 5565. The strain can normally grow under the extreme alkaline environment of pH11, shows a good oligotrophy-tolerant characteristic, can degrade a variety of hemicellulose-containing biomasses including corn straws, rice straws, Jerusalem artichoke straws, kelp fibers and the like, and has a broad application prospect in utilizing cheap biomass resources to produce bio-based chemicals. The strain can produce xylanase under optimum conditions, and the enzymatic activity of the xylanase can be as high as 68.9U/ml. The produced xylanase has the following properties: the optimum pH value is 6.0, the optimum temperature is 70 DEG C, the optimum substrate is beech xylan, and the produced xylanase does not have cellulase activity, has high pH stability and thermal stability, and can tolerate high-concentration NaCl. As an enzyme preparation coming from the ocean, the xylanase can be widely used in papermaking, food, feed, wine making, energy and other industries.
Description
Technical field
The present invention relates to a strain and produce ocean green product look streptomycete (M11) bacterial strain and the application thereof of alkaline-resisting salt tolerant zytase, be specially adapted to the alkaline-resisting salt tolerant zytase of large-scale industrial production, belong to biological technical field.
Background technology
Utilizing cheap biomass resource to produce high value added product is the research content of biotechnology.Xylan is a kind of abundant renewable resources of occurring in nature, is the most representative hemicellulose components, accounts for the 1/3-1/2 of semicellulose, is the abundantest polysaccharide of occurring in nature except that Mierocrystalline cellulose.Therefore, how to utilize xylan to become at present the focus of research both at home and abroad.
Xylan is (with β-1 by D-wood sugar main chain; 4 keys link to each other) and L-arabinose branch (α-1; 2 and α-1,3 link to each other) polymkeric substance formed, link to each other with 4-O-methyl-D-glucuronic acid key with Mierocrystalline cellulose with xylogen; Formed the structure of crosslinked complicacy, for degradation of hemicellulose is brought difficulty.At present, the method that industrial degradation of xylan is commonly used has acid system, alkaline process and enzyme process, and wherein acid system, alkaline process are used comparatively extensively, but contain more toxic substance in the acid and alkali hydrolysis liquid, for the later stage microbial fermentation restraining effect are arranged; In addition, adopt acid system or alkaline process degradation of xylan, it is very big to produce environmental protection pressure, therefore receives the restriction of country, so from the long-term production effect, the enzymic degradation xylan will be the main mode of following xylan degrading.
Zytase is the lytic enzyme of most critical in the xylan hydrolysis enzyme system.When xylan was degraded, the enzyme that plays a major role was β-1,4-endo-xylanase and β-D-xylosidase.β-1, the 4-endo-xylanase acts on inner β-1, the 4 wood sugar glycosidic bond of xylan backbone with internal-cutting way, and its main hydrolysate is xylooligosaccharides, wood oligose, xylo-bioses etc.; β-D-xylosidase comes catalysis to discharge xylose residues through the non reducing end of hydrolysis xylooligosaccharides, wood oligose etc.
Zytase has huge using value and wide application prospect in various fields such as fodder industry, foodstuffs industry, paper industry, energy industries.For example, in the starting stage of association with pulp bleaching, in paper pulp, adding zytase can increase the luminance brightness and the degree of hydrolysis of paper pulp, thereby has reduced muriatic consumption in the later stage bleaching, has reduced the pollution to environment; In the bread baking process, in dough, add zytase, can shorten the fermentation time of dough, improve the elasticity and the hardness of crumb, increase the volume and the specific volume of bread, improve the bread baking quality effectively; In feed, add the production performance that zytase can significantly improve livestock and poultry.Zytase can destroy the vegetable cell wall construction, improves the utilization ratio of all kinds feed, reduces livestock and poultry intestinal canal diseases, promotes livestock birds health, improves the livestock and poultry surviving rate; Reduce sticking excrement, reduce ammonia and sulfide concentration in the air, reduce environmental pollution, reduce sticking excrement and discharge and dirty egg, make the livestock and poultry body weight even.Therefore, be accompanied by human attention for sustainable development and environment, the application of zytase in industry will receive people's attention more.
Marine actinomycete mainly is distributed in marine bottom sediment, sea life surface and the seawater.The singularity of ocean environment has brought up unique metabolic way and the meta-bolites of marine actinomycete.People constantly search out from various ocean environment and can produce microbiotic, zymin (proteolytic enzyme, glycase and cellulase etc.), VITAMINs (B12) and the organic acid actinomycetes with novel mechanism of action in recent years.In the limit of life environment of oceanic high, low temperature, high salt, marine actinomycete has unique pathways metabolism and synthesis capability, and this not only guarantees it and in extreme environment, survives, and the potentiality that produce special meta-bolites also are provided.The enzyme of marine microorganism production generally has the characteristic relevant with its living environment, for example, salt tolerance, high thermal resistance, suitable to cold etc., these characteristics have broad prospect of application under industrial extreme condition.In addition, actinomycetic genetic manipulation is simple, is convenient to carry out genetic modification, compares with mould to have more biological safety.
Summary of the invention
The object of the present invention is to provide a strain to produce ocean green product look streptomycete (M11) bacterial strain and the application thereof of alkaline-resisting salt tolerant zytase, this bacterial strain has oligotrophic, pH, NaCl tolerance character, can be used for the alkaline-resisting salt tolerant zytase of fermentative prepn; Unique zymologic property that above-mentioned alkaline-resisting salt tolerant zytase is had makes it and can be widely used in the industrial production.
The present invention relates to a strain and produce green look streptomycete (M11) bacterial strain that produces in ocean of alkaline-resisting salt tolerant zytase; Classification called after Streptomyces viridochromogenes; Registering on the books of said bacterial strain is numbered CGMCC No.5565, is preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center, and the depositary institution address is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City; Institute of Microorganism, Academia Sinica, preservation date is on December 9th, 2011.
Green look streptomycete (the following omission M11) bacterial strain that produces in said ocean can be produced alkaline-resisting salt tolerant zytase, is adapted at using in papermaking, feed, food, wine brewing and the energy industry.
When said bacterial strain was grown on the Bennett solid medium, bacterium colony was circular, and densification goes out lime look spore through bacterium colony surface growth in 2-3 days, and substrate is blackish green, produces black pigment, and the speed of growth is very fast.This bacterial strain has oligotrophic property, pH, NaCl tolerance preferably, is that bacterial strain is well-grown still under 11 the extreme alkaline condition in the pH value; Growth is best in 1% NaCl.Bacterial strain of the present invention can be grown in the substratum that with the xylan is sole carbon source, and the fermented liquid that is produced has the ability of degradation of xylan.The fermenting enzyme that ferments 7 days the time is lived the highest, can reach 68.9U/ml.
The zytase that said bacterial strain is produced has characteristics such as pH stability, NaCl tolerance and thermostability preferably simultaneously.The zytase that the green color-producing streptomycete that the present invention screens is produced, its optimum pH are 6.0, in the scope of pH5-9, keep the enzymic activity more than 80%; Optimum temperuture is 70 ℃, handles 60min down at 45 ℃, and enzymic activity maintains more than 88%; Handle 60min down at 60 ℃, enzymic activity maintains more than 75%; The righttest substrate of zytase among the present invention is the beech wood glycan, the cellulose-less enzymic activity; This zytase has good NaCl tolerance, and when the NaCl final concentration was 1mol/L, xylanase activity maintained 86.6%; When the NaCl final concentration was 2M, residual enzyme lived 70.4%; When the NaCl final concentration was 5mol/L, residual enzyme lived 46.8%.The described zytase with these characteristics of this patent does not also appear in the newspapers.
Zytase degradable various agricultural waste and cheap biomass that said bacterial strain is produced, best to the degradation effect of jerusalem artichoke stalk.In addition, the sea-tangle fiber of marine source is also had degradation property preferably, show that this zytase has using value aspect the high value product of ocean kelp production utilizing.
Said zytase is 9 o'clock at pH; Residual enzyme is lived and is 83.3% of maximum enzyme activity; Showed alkali resistance preferably, and kept higher enzyme activity down, can satisfy the bio-bleaching technology in the papermaking at 60 ℃; Semicellulose in the hydrolysis paper pulp increases paper quality, hardness, quality to a certain extent.The more important thing is that replacement or part have reduced muriatic usage quantity, reduce the pollution to environment.Zytase also can be applicable to wine industry among the present invention, reduces material viscosity, improves the utilization ratio of starch, increases the productive rate of alcohol.Zytase among the present invention also demonstrates huge application potential in energy industry; This enzyme can be kept high enzyme for a long time and live under 45 ℃; Consistent with present synchronous saccharification technique temperature; And the semicellulose in the degradable agricultural wastes is a monose, supplies the downstream process fermentation to utilize, and produces clean reproducible energy.
The invention has the beneficial effects as follows: this strains separation was preserved in Chinese common micro-organisms DSMZ on December 9th, 2011 in the ooze sample of Dalian, preserving number is 5565.This bacterial strain is normal growth under 11 the extreme alkaline environment in the pH value; And showed the oligotrophic characteristic of good tolerance; The multiple biomass that contain semicellulose of ability degraded; Comprise corn straw, straw, jerusalem artichoke stalk, sea-tangle fiber etc., have broad application prospects aspect the cheap biomass resource production bio-based chemical utilizing.This bacterial strain is produced zytase under optimum condition, xylanase activity reaches as high as 68.9U/ml.The zytase of producing has following character: optimum pH 6.0, and 70 ℃ of optimum temperutures, the righttest substrate is the beech wood glycan, the zytase cellulose-less enzymic activity of producing has good pH stability, thermostability, and can tolerate high density NaCl.As a kind of zymin of marine source, can be widely used in industry such as papermaking, food, feed, wine brewing, the energy.
Description of drawings
Fig. 1 is the green look streptomycete oligotrophic tolerance of producing in ocean.A. contrast; B. dilute 2 times; C. dilute 3 times; D. dilute 5 times.
Fig. 2 is the green look streptomycete pH tolerance of producing in ocean.a.pH＝5；b.pH＝7；c.pH＝9；d.pH＝11。
Fig. 3 is the green look streptomycete NaCl tolerance of producing in ocean.A. contrast; B.1%NaCl; C.2%NaCl; D.3%NaCl.
Fig. 4 is that the green look streptomycete liquid fermenting that produces in ocean is produced the zytase result.
Fig. 5 is the green look streptomycete zytase optimum temperuture result that produces in ocean.
Fig. 6 is the green look streptomycete zytase thermostability result that produces in ocean.
Fig. 7 is the green look streptomycete zytase ph optimum result that produces in ocean.
Fig. 8 is the green look streptomycete zytase pH stability result of producing in ocean.
Fig. 9 is the green look streptomycete zytase NaCl tolerance result that produces in ocean.
Embodiment
Experiment material and reagent
1. bacterial strain: the ocean is green produces the look streptomycete and from the Xiao Pingdao ooze sample of Dalian, was separated in 2006 by the inventor and obtain.
2. biochemical reagents:
Xylan: beech wood glycan, birch xylan, oat xylan (U.S. SIGMA company);
The sea-tangle fiber is available from the kind edge in Shaanxi Bioisystech Co., Ltd.
Sepharose DNA reclaims test kit (Solarbio company);
Order-checking is accomplished by the precious biotechnology in Dalian ltd;
TaKaRa rTaq Dalian precious biotechnology ltd;
10xPCR Buffer Dalian precious biotechnology ltd;
DNTP Mixture Dalian precious biotechnology ltd.
Extract strain gene group DNA reagent:
Washings: 50mmol/L Tris, 25mmol/L EDTA, 0.1%PVP, pH are 8.0.
Lysate: 50mmol/L Tris, 25mmol/L EDTA, 1.2%PVP, 3%SDS.
Extract liquid: 10mmol/L Tris, 1mmol/L EDTA, 0.3mmol/L NaAC, pH are 8.0.
Extracting solution: phenol: chloroform: primary isoamyl alcohol=25: 24: 1.
The TE damping fluid: 50mmol/L Tris, pH are 8.0.
The DNS agent prescription:
First liquid: dissolving 6.9g crystalline phenol adds 6.9g NaHSO with distilled water diluting to 69mL again in the NaOH of 15.2mL 10%
3
Second liquid: take by weighing the 255g Seignette salt, be added among the 10%NaOH of 300mL, again to wherein adding 880mL1%3,5-dinitrosalicylic acid solution;
First and second liang of liquid phases are mixed promptly obtaining yellow DNS reagent, be stored in the brown reagent bottle.At normal temperatures, place and use after 7-10 days, effective in half a year.
3. substratum
Bennett solid medium: glucose 10g, peptone 2g, yeast powder 1g, Carnis Bovis seu Bubali cream 1g, agar 20g, pH7.0.
Xylan solid medium: beech wood glycan 10g, NH
4NO
34g, K
2HPO
43H
2O 1g, MgSO
47H
2O 0.5g, NaCl 0.3g, agar 20g, pH6.5-7.0.
Enrichment medium: (NH
4)
2SO
45g, KH
2PO
41g, MgSO
47H
2O 0.5g, FeSO
47H
2O 0.01g, CaCl
20.2g, beech wood glycan 10g, pH 7.0.
TSB liquid nutrient medium: Tryptones 17g, soy peptone 3.0g, glucose 2.5g, K
2HPO
43H
2O 2.5g, NaCl 5g, pH 7.0.
Zytase produces enzyme substratum: K
2HPO
43H
2O 1.11g, KH
2PO
43.65g, (NH
4)
2NO
33g, MgSO
47H
2O 2.2g, CaCl
20.3g, glucose 1.5g, peptone 2.5g, Carnis Bovis seu Bubali cream 3g, beech wood glycan 10g, pH 6.5.
Explain: make the experimental methods of molecular biology specify in following examples, all carry out, perhaps carry out according to test kit and product description with reference to the listed concrete grammar of " molecular cloning experiment guide " (third edition) J. Sa nurse Brooker one book.
Embodiment 1: the acquisition of bacterial strain
To derive from Dalian Xiao Pingdao ooze sample after enrichment culture; Coat on the xylan flat board by after the routine dilution, cultivated 5-6 days for 30 ℃, picking produces the separation of on the xylan solid medium, ruling of transparent circle bacterium colony; The sepn process 3 that repeats to rule is taken turns, and makes the bacterial strain purifying.Screen the bacterial strain of this secretion zytase through this method.
Bacterial strain of the present invention was cultivated 7 days under 30 ℃ on the Bennett flat board, and bacterium colony is rounded, and diameter is about 3-4cm, and substrate is blackish green, and the bacterium colony protrusion of surface produces lime look spore, discharges a large amount of black pigments.Its ph optimum is 7.0-9.0, and optimum temperuture is 30 ℃.
Embodiment 2: extract the green look streptomyces gene group DNA that produces in ocean, with and the acquisition of 16SrDNA
Extract the green look streptomyces gene group DNA that produces in ocean:
1) with inoculation in the TSB of 50mL liquid nutrient medium, cultivated 72 hours at 30 ℃ of following rotating speed 170rpm, be in logarithmic phase this moment.Collect fermented liquid 1mL, abandon supernatant behind the centrifugal 5min of 8000r/min, washings cleans thalline, repeats once to clean after centrifugal.
2) in the thalline after washing, add 100 μ L lysate suspension thalline, 100 ℃ are boiled the 15min postcooling to room temperature.
3) add 65 ℃ of extract 0.5ml that preheating is good, slight concussion.
4) add the saturated phenol/chloroformic solution of equal-volume Tris, shake 1min strongly, the centrifugal 5min of 10000r/min carefully draws supernatant, is transferred to clean centrifuge tube.
5) in supernatant, add isopyknic Virahol, the reversing mixing was left standstill 2 hours for twice ,-20 ℃ gently.
6) the centrifugal 5min of 12000r/min, 70% washing with alcohol once, the centrifugal 5min of 10000r/min, opening evaporate into does not have the alcohol flavor, is dissolved in after the drying in the 20 μ LTE damping fluids, makes total DNA.
The acquisition of 16SrDNA sequence:
16SrDNA PCR primer is following:
A P1:5 ' ATTCGGCACACAGAAAC 1 '
A P2:5 ' AGAGGAGAACCGIAGAC 1 '
The PCR reaction conditions is as shown in table 1:
Table 1PCR reaction conditions
The PCR reaction system is as shown in table 2:
Table 2PCR reaction system
Embodiment 3: green oligotrophic tolerance, pH tolerance and the NaCl tolerance of producing the look streptomycete in ocean
With 1,2,3,5 times of Bennett solid medium dilution, in order to measure the green oligotrophic tolerance of producing the look streptomycete in ocean; Regulating Bennett solid medium pH is 5,7,9,11, to measure the pH tolerance of bacterial strain; In the Bennett solid medium, add 0%, 1%, 2%, 3% NaCl, to measure the NaCl tolerance of bacterial strain.(concentration is about 10 to get 50 μ L spore solution
6Individual/as mL) evenly to coat above-mentioned each flat board respectively, cultivated 6 days down in 30 ℃, observe the colony growth state.Result's (seeing Fig. 1-3) shows bacterial strain well-grown on the Bennett flat board of 5 times of dilutions, and its oligotrophic property tolerance preferably is described; Bacterial strain is grown relatively poor on the flat board of pH 5, is still can normal growth on 11 the flat board at pH, explains that bacterial strain alkali resistance of the present invention is stronger, and the righttest growth pH is 7-9; Bacterial strain is grown best on 1% NaCl flat board, and pigment production is maximum, shows that bacterial strain of the present invention has salt tolerance preferably.
Embodiment 4: ocean green product look streptomycete carries out liquid fermenting and produces zytase in producing the enzyme substratum
The streak inoculation ocean is green produces the look streptomycete on the Bennett solid medium, cultivates 3-5 days down at 30 ℃.When treating the spore well-grown, with the rifle of 1000 μ L end to end nose circle get 4 ferfas and fall to being inoculated in the TSB liquid nutrient medium, cultivate 48h in 30 ℃ of following rotating speed 170rpm, carry out seed culture.Draw the cultured seed substratum, the inoculum size with 5% inserts the 50mL zytase and produces in the enzyme substratum, is 170rpm at rotating speed, and temperature is to cultivate under 30 ℃ of conditions.Result (Fig. 4) shows, ferments that zytase output reaches the highest after 7 days, and enzyme work is about 68.9U/mL.
Embodiment 5: measure xylanase activity in the green product look streptomycete fermentation liquid of ocean
With the centrifugal 10min under 8000rpm of the fermented liquid among the embodiment 3, collect supernatant as crude enzyme liquid.Crude enzyme liquid is accurately drawn crude enzyme liquid 0.2mL (3 parallel test tubes of each sample) through suitably dilution, adds substrate (1% beech wood glycan) 1.8mL.Mix the back and in 60 ℃ of water-baths, react 5min, reaction finishes the back and adds DNS liquid 3.0mL termination reaction rapidly, in boiling water, boils 5min then, takes out postcooling to room temperature.Add zero(ppm) water to 25mL, shake up.Measure the OD value down in 540nm, according to wood sugar typical curve conversion wood sugar content.Blank, substrate 1.8mL add DNS reagent 3.0mL after in 60 ℃ of water-baths, handling 5min, and to the crude enzyme liquid that wherein adds after 0.2mL dilutes, other processing is consistent with sample again.With the blank is reference working sample absorbance A.1 enzyme unit alive (U) is defined as under above-mentioned condition, and PM discharges the required enzyme amount of 1 μ mol wood sugar.
Embodiment 6: the green mensuration of producing look streptomycete zytase zymologic property in ocean
1. green optimum temperuture and the thermostability of producing look streptomycete zytase in ocean
Enzymatic reaction is carried out in being determined as under phosphate buffered saline buffer (pH6.0) system and differing temps of the optimum temperuture of zytase.Temperature tolerance is determined as zytase and handles down 10-60min at 45,60,70 ℃ respectively, be cooled to room temperature after, under 60 ℃, carry out enzyme assay again.The enzyme reaction optimum temperuture is measured result (Fig. 5) and is shown that its optimum temperuture is 70 ℃.The thermostability experimental result of zytase shows (Fig. 6), and after handling 60min under 45 ℃, enzyme is lived and all remained on more than 88%; After handling 60min under 60 ℃, enzyme is lived residual more than 75%.
2. the ph optimum and the pH of the green product in ocean look streptomycete zytase are stable
The crude enzyme liquid of dilution among the embodiment 3 is carried out enzymatic reaction to measure its ph optimum under different pH.The beech wood glycan is with the enzyme reaction buffer solution configuration of different pH, 50mM citrate buffer 4.0-6.0,50mM phosphate buffered saline buffer 6.0-8.0,50mM Tris-HCl damping fluid 8.0-9.0.Under 60 ℃, carrying out xylanase activity measures.Result (Fig. 7) shows that the ph optimum of zytase that bacterial strain of the present invention produces is 6.0, and in the scope of pH5-8, enzymic activity all surely is held in more than 40% of maximum enzyme activity.Crude enzyme liquid in the damping fluid of above-mentioned various different pH 50 ℃ handle 30min, be cooled to room temperature after acid base titration regulate pH to 6.0, again in the pH6.0 buffer solution system 60 ℃ measure enzymic activity down, with the pH stability of research zytase.Result (Fig. 8) shows, zytase is stable in the scope of pH5-9, is that residual xylanase activity explains that 83.3% of maximum enzyme activity this enzyme has alkali resistance preferably under 9 the condition at pH.
3. produce the influence that the righttest substrate of enzyme and different metal ion and chemical reagent are lived to enzyme
Be substrate with 1% beech wood glycan, birch xylan, oat xylan, filter paper respectively, under pH6.0,60 ℃ of systems of temperature, measure xylanase activity.Result's (seeing table 3) shows that the righttest substrate of zytase that bacterial strain of the present invention produces is the beech wood glycan, and agricultural wastes corn cob xylan is also had good degradation property, this zytase cellulose-less enzymic activity.In enzymatic reaction system, add the different metallic ion and the chemical reagent of different concns, study its influence to enzymic activity, various material final concentrations are 1mmol/L and 10mmol/L.At 60 ℃, measure enzymic activity under the pH6.0 condition.Result's (table 4) shows, the Mn of lower concentration
2+, SDS and EDTA have certain restraining effect to xylanase activity; The Zn of high density
2+Work has certain restraining effect to enzyme with EDTA, the Cu of high density
2+, Fe
2+Work has had strong inhibitory effects to enzyme with SDS.The Ca of high density
2+, Mg
2+, K
+Plasma is lived influence not quite to enzyme.
4. the green NaCl tolerance of producing look streptomycete zytase in ocean
In enzymatic reaction system, add NaCl, final concentration is 0.5-5mol/L, studies its influence to xylanase activity, at 60 ℃, measures enzymic activity under the pH6.0 condition.Result (Fig. 9) shows that when the NaCl final concentration was 1mol/L, xylanase activity maintained 86.6%; When the NaCl final concentration was 2M, residual enzyme lived 70.4%; When the NaCl final concentration was 5mol/L, residual enzyme lived 46.8%.Under extreme hypersaline environment, zytase that bacterial strain of the present invention produces still keeps the high enzyme vigor, explains that this zytase has NaCl tolerance preferably.
5. the green degraded of producing look streptomycete zytase in ocean to cheap biomass
With the jerusalem artichoke stalk, corn straw, straw are crossed 60 mesh sieves after crushed, handle 1h with 1: 12 usefulness 4% dilute sulphuric acid of solid-to-liquid ratio down at 121 ℃, are cooled to use after the room temperature deionized water to wash repeatedly to pH to be neutrality.After the dried overnight, take by weighing above-mentioned material of 1g and sea-tangle fiber respectively and be dissolved in 20mL50mM phosphate buffered saline buffer (pH6.0).Behind 60 ℃ of preheating 30min, add crude enzyme liquid with the 50U/g ratio, 60 ℃ are reacted 1h down, and reaction finishes the back and draws supernatant, press method mensuration concentration of reduced sugar among the embodiment 4.Result's (table 5) shows that zytase that bacterial strain of the present invention produces has degradation property preferably to jerusalem artichoke stalk and sea-tangle fiber.Contain a certain proportion of NaCl in the sea-tangle fiber of marine source, the zytase among the present invention has NaCl tolerance preferably, can efficient degradation sea-tangle fiber.Zytase among this result of study explanation the present invention is having broad application prospects aspect the high-valued bio-transformation of biomass resource.
The green the righttest substrate of look streptomycete zytase that produces in table 3 ocean
Attach: the enzyme work when being substrate with the beech wood glycan is 100%.
Table 4 metals ion and chemical reagent are to the green influence of producing look streptomycete zytase in ocean
Attach: the sample enzyme work not add ion processing is 100%.
The green degraded of producing the look streptomycete in table 5 ocean to cheap biomass
Attach: with the degradation rate to the jerusalem artichoke stalk is 100%.
Claims (2)
1. the green look streptomycete bacterial strain (Streptomyces viridochromogenes) that produces in ocean of alkaline-resisting salt tolerant zytase is produced in a strain; It is characterized in that: registering on the books of said bacterial strain is numbered CGMCC No.5565; Be preserved in Chinese culture presevation management committee common micro-organisms center, preservation date is on December 9th, 2011.
2. the green application of producing the look streptomycete bacterial strain in ocean of alkaline-resisting salt tolerant zytase is produced in a strain according to claim 1, it is characterized in that: the green look streptomycete bacterial strain that produces in said ocean is applied to the alkaline-resisting salt tolerant zytase of industrial production.
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| CN103215240A (en) * | 2013-02-26 | 2013-07-24 | 云南大学 | Two Ag<+> and SDS resistant xylanases |
| CN104263667A (en) * | 2014-06-13 | 2015-01-07 | 湖北工业大学 | Streptomyces althioticus and applications thereof |
| CN104450584A (en) * | 2014-12-12 | 2015-03-25 | 杨凌农科大无公害农药研究服务中心 | Actinomycetes and application thereof |
| CN104560915A (en) * | 2013-10-11 | 2015-04-29 | 镇江拜因诺生物科技有限公司 | A high-density fermentation production method of alkaline xylanase |
| CN106459941A (en) * | 2014-04-01 | 2017-02-22 | 韩国生命工学研究院 | Novel xylanase produced from streptomyces sp. strain hy-14 |
| CN108220189A (en) * | 2017-12-29 | 2018-06-29 | 厦门大学 | One plant of marine bacteria for producing zytase and application |
| CN108277176A (en) * | 2018-01-31 | 2018-07-13 | 深圳大学 | Alkalescent xylanase and the application of a kind of basophilic streptomycete and its generation |
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| CN103215240A (en) * | 2013-02-26 | 2013-07-24 | 云南大学 | Two Ag<+> and SDS resistant xylanases |
| CN103215240B (en) * | 2013-02-26 | 2014-04-02 | 云南大学 | Two Ag<+> and SDS resistant xylanases |
| CN104560915A (en) * | 2013-10-11 | 2015-04-29 | 镇江拜因诺生物科技有限公司 | A high-density fermentation production method of alkaline xylanase |
| CN106459941A (en) * | 2014-04-01 | 2017-02-22 | 韩国生命工学研究院 | Novel xylanase produced from streptomyces sp. strain hy-14 |
| CN106459941B (en) * | 2014-04-01 | 2020-01-10 | 韩国生命工学研究院 | Novel xylanase produced from streptomyces strain HY-14 |
| CN104263667A (en) * | 2014-06-13 | 2015-01-07 | 湖北工业大学 | Streptomyces althioticus and applications thereof |
| CN104450584A (en) * | 2014-12-12 | 2015-03-25 | 杨凌农科大无公害农药研究服务中心 | Actinomycetes and application thereof |
| CN108220189A (en) * | 2017-12-29 | 2018-06-29 | 厦门大学 | One plant of marine bacteria for producing zytase and application |
| CN108277176A (en) * | 2018-01-31 | 2018-07-13 | 深圳大学 | Alkalescent xylanase and the application of a kind of basophilic streptomycete and its generation |
| CN108277176B (en) * | 2018-01-31 | 2020-12-04 | 深圳大学 | Alkaliphilic streptomyces, alkaline xylanase produced by same and application of alkaline xylanase |
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