CN103667151A - Bacillus thermophilus capable of producing high-temperature and alkali resistant xylanase and application of bacillus thermophilus - Google Patents
Bacillus thermophilus capable of producing high-temperature and alkali resistant xylanase and application of bacillus thermophilus Download PDFInfo
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Abstract
The invention discloses a bacillus thermophilus 701 capable of producing high-temperature and alkali resistant xylanase and belongs to the technical field of microbial fermentation. The bacillus thermophilus is obtained through the steps that a soil sample is collected from a paper mill, a bacillus thermophilus is separated from the soil sample, and ultraviolet mutagenesis and nitrosoguanidine mutagenesis, and screening are performed repeatedly. The xylanase produced by the bacillus thermophilus has the characteristics of strong high-temperature resistance and alkali resistance. The bacillus thermophilus is preserved in the China Center for Type Culture Collection on October 12, 2013, the preservation No. is CCTCC M2013537. The high-temperature and alkali resistant xylanase prepared from the bacillus thermophilus 701 is 350-420 IU/ml in specific activity, 40-75 DEG C in applicable temperature range, 65 DEG C in optimum reaction temperature, completely stable in enzyme activity at the temperature of 75 DEG C, 5.0-11.0 in applicable reaction pH value, completely stable in enzyme activity at the pH value of 11.0, and is 10.0 in optimum reaction pH value.
Description
Technical field
The invention belongs to microbial fermentation technology field, specifically a kind of bacillus acidocldarius and application thereof of producing high temperature resistant alkali body acidic xylanase.
Background technology
Mierocrystalline cellulose, hemicellulose and xylogen are the main components of plant hemicellulose, their actings in conjunction have formed the support frame of plant cell wall, among three, hemicellulose has occupied the more than 35% of plant dry weight, hemicellulose is also the general name of multiple complex plycan, its main component is xylan, and it is a kind of poly five-carbon sugar of complexity.The main chain of xylan is by β-1-4 glycosidic link, a plurality of xylopyranosyl to be coupled together, and is connecting different substituting groups on side chain, has like this (Collins et al., 2005) such as ethanoyl, glucal acyl group, L-arabinose bases.So degradable common participation that just needs plurality of enzymes of xylan.Generally xylan is divided into hardwood xylan and cork xylan.Hardwood xylan is to be mainly polymerized by 0-acetyl-4-0-methyl glucoside aldehyde acyl wood sugar, with β-1-4 glycosidic link, is connected.Cork glycan is by Arabic 4-0-methyl glucoside aldehyde acyl xylose residues be polymerized (Beg et al, 2001).Some sorting techniques are also divided into soluble xylan and insolubility xylan xylan.
Zytase (Isosorbide-5-Nitrae-β-D-xylanase:EC3.2.1.8) is a kind of important industrial enzymes, when decomposing xylan, plays biocatalysis, xylan can be resolved into protein or the RNA of polysaccharide or monose.Zytase is extensively present in natural organism.Can produce zytase in bacterium, fungi, animal body etc.
Because zytase is in industrial potential using value, caused within the past ten years more attentions of people, wherein alkalescent xylanase is mainly used in paper pulp and papermaking enterprise.The application of alkalescent xylanase in paper industry mainly comprises slurrying, assists bleaching, changes the aspects such as fibrous property, deinking.
In industrial production, pulping process is under alkaline condition, by the method for high temperature steaming, remove delignification, for making to dissolve pulping fibre element purity, reach 98%, this just needs a large amount of sodium-hydroxide treatment paper pulp, caused serious environmental pollution, many scholars slurrying then trial is carried out a biological disposal upon for this reason, and should be heat-resisting and alkaline-resisting for the zytase of pulping bleaching.Feng Jianliang etc. (Kimura et al.2000) compare test with xylanase pretr eatment wheat straw and the slurrying of conventional chemical method, and xylanase pretr eatment has improved the delignification degree of raw material, can also reduce the Kappa number of slurrying.Zytase has compared with extensive and deep research aspect auxiliary bleaching, and has obtained very large achievement.Research is found, no matter is softwood pulp, hardwood pulp or bamboo pulp, straw pulp, and the zytase remaining xylogen in paper pulp of all can degrading, has improved whiteness (AL BALAA et al, 2006; Meek and Lipman, 1922).Zytase for association with pulp bleaching must have high temperature resistant feature, Khasin etc. have obtained one and have derived from alkaline bacterial strain Bacillus sterarothermophilus T26 zytase, this zytase has best bleaching effect (Khasin et al to paper pulp when pH9.0 and 65 ℃, 1993), a lot of zytases do not possess such feature, have limited commercial applications.The annual waste paper quantity producing in the whole world is surprising, and after waste paper reclaims, recycling work just becomes particularly important, can make also to have protected environment in resource recycling.1991, by Korea S scholar reported first application zytase ink can be removed from news paper waste.After this, people begin one's study and utilize zytase to carry out deinking, the environmental pollution (Cao Junwei etc., 2004) bringing to alleviate or to eliminate Chemical Deinking.
At present, the green cleaning and bleaching of paper pulp that element-free chlorine and Totally-chlorine-free bleaching technology be representative of take has become the inexorable trend of countries in the world paper-making industry association with pulp bleaching development, zytase enzyme process helps and floats novel process and in Europe and more than 30 large-scale paper plants of North America, be applied, and becomes biotechnology and applies the most successful example in paper industry.Wherein have an appointment 10% sulphate process pulp mill of Canada has adopted this novel process.Denmark Novozymes Company and U.S.'s mountain pass Deng Duojia of this chemical company zymin manufacturer, released one after another and be specifically designed to zytase and the cellulase product innovation that slurrying is processed, but up to the present, the industrial zytase that is applied to association with pulp bleaching is neutral meta-acid mostly, optimal reactive temperature is mostly 50 ℃ of left and right, as everyone knows, pulp cooking and bleaching are substantially all to carry out under the condition of high temperature and highly basic, make the application of existing low temperature acidic xylan enzyme product in this field be subject to great restriction, in addition, at feed and field of food, zytase is applied in the granulation of feed and baking in operation of food, still require zytase used under hot conditions, can keep higher enzyme to live, therefore, research and develop resistant to elevated temperatures zytase product and will bring good economic benefit and social benefit.
The xylanase bacterium that sifting property is more good from natural microbial is means the most effective that effectively obtain high yield zytase bacterial strain.Since zytase has bio-bleaching function, people have invested alkalescent xylanase more sight, have screened the production bacterial strain of the alkalescent xylanase much with application prospect, separated a lot of alkalescent xylanases.1973, Horikoshi etc. found that purifying obtains zytase from alkaline bacterium Bacillus sp.No.C25922 from the zytase in alkaline bacterium (Horikoshi and Atsukawa, 1973) first, and its optimal pH is 6-8.Afterwards, 2 zytases from alkaline bacterium Bacillus No.C2125 have been found: zytase A and zytase N, wherein zytase A also has certain enzyme (Honda et al, 1985) alive when pH12.The basophilic thermophile bacteria Bacillus sp. (NCIM59) that Dey etc. (Dey et al, 1992) are separated, can produce 2 zytases that there is no cellulase activity, and they have good alkali resistance, and optimal pH is 10.0.Nakamura etc. have reported that outside the born of the same parents that derive from alkaline bacterial strain Bacillus sp.41M21, its optimal pH of zytase J is 9.0(Nakamura et al, 1993).Most of mycetogenetic zytases are generally acidic xylanase, but also there is exception, Taneja etc. screen a strain basophilic fungi AspergillusnidulansKK299, the optimal pH of the zytase that it produces is 8(Taneja et al, 2002), and this enzyme hardwood xylan is had to higher affinity than cork xylan.
Chinese patent CN101701205A discloses a kind of alkali resistance zytase XynE2 and gene and application, the alkali resistance zytase optimum pH 7.8 of producing, 65 ℃ of optimum temperutures; Chinese patent CN102586134B discloses green look streptomycete bacterial strain and application thereof, the zytase optimum pH 6.0 of producing, 70 ℃ of the optimum temperutures of producing in ocean that alkaline-resisting salt tolerant zytase is produced in a strain; Chinese patent CN102321558A discloses a kind of high-yield strains of fire resistant xylanase and has utilized this bacterium fermentation to produce the method for fire resistant xylanase and the enzyme obtaining, the fire resistant xylanase optimum pH 7.0 of producing, 60 ℃ of optimum temperutures.
In view of this, international and domestic all multi-experts, scholar and Scientific Research Workers are not stinted fund, manpower and time, strengthened R&D intensity high temperature resistant, alkali resistance zytase, be intended to by biotechnology the application in the fields such as papermaking, food, feed push to one higher, upgrade, stronger New Times, no matter be the acquisition of novel bacterial, still adopt gene recombination technology, although expended big financial resources like this and energy, but result is unsatisfactory, be difficult to reach R&D target and requirement high temperature resistant, alkali resistance zytase, or heatproof is not alkaline-resisting; Alkaline-resisting not heatproof; Alkaline-resisting high temperature resistant scope is not wide in range; Zytase is impure and have a cellulase activity; Effect substrate discomfort etc., a kind of strain excellent that produces high temperature resistant alkali body acidic xylanase of isolation and selection is a kind of necessity.
Summary of the invention
The object of this invention is to provide a kind of bacillus acidocldarius (Bacillus thermophilus) 701 that produces high temperature resistant alkali body acidic xylanase.
The Xue Li paper mill, bacillus acidocldarius 701Shi Cong Hunan Province of product high temperature resistant alkali body acidic xylanase provided by the invention gathers soil sample and isolates a strain bacillus acidocldarius HYX0021, through ultraviolet mutagenesis repeatedly and nitrosoguanidine mutagenesis screening, obtain, characteristic be high temperature resistant, alkali resistance is strong.
The bacterial strain of product high temperature resistant alkali body acidic xylanase provided by the invention is specially bacillus acidocldarius (Bacillus thermophilus) 701.This bacterial strain is preserved in Chinese Typical Representative culture collection center on November 3rd, 2013 and (is called for short CCTCC, address is: Wuchang District, Wuhan City, Hubei Province Luo Jia Shan Wuhan University Life Science College, postcode: 430072), preserving number is CCTCC NO:M2013537, and Classification And Nomenclature is bacillus acidocldarius (Bacillus thermophilus) 701.
Bacillus acidocldarius 701 provided by the invention is Gram-positives, and bacillus is aerobic, produces gemma, on LB solid medium, cultivates 24h for 37 ℃, and bacterium colony is rounded, and colony diameter is about 1.0-2.0 ㎜, oyster white, and smooth surface, opaque, the irregular expansion in edge.Peritrichous, can move, without pod membrane, have gemma, thalline size is (0.7～0.9) μ m * (3～3.5) μ m, gemma (0.6～0.9) μ m * (1.0～1.5) μ m, ellipticity, is positioned at thalline central authorities or slightly inclined to one side, and after sporulation, thalline does not expand.While growing in liquid nutrient medium, form wrinkle mould.Casein hydrolysis experiment, Starch Hydrolysis experiment, gelatine liquefication experiment, nitrate reduction are tested all positive; Thalline can utilize glucose, fructose, sucrose, can not utilize lactose and N.F,USP MANNITOL; H
2s, catalase experiment, indole test are tested all negative.
Bacillus acidocldarius 701 provided by the invention carries out 16sRNA gene sequencing, resulting sequence is carried out to homology comparison by existing 16SrDNA sequence in Blast software and Genbank, utilize software MEGA4.0 to build evolutionary tree, result shows that it is one that bacterial strain of the present invention and bacillus acidocldarius (Bacillus thermophilus) gather, sequence similarity is 98%, the morphological feature of comprehensive 701 bacterial strains, physiological and biochemical property and 16S rDNA nucleotide sequence homology analytical results, bacterial strain 701 is accredited as to bacillus acidocldarius (Bacillus thermophilus).
The prepared high temperature resistant alkali body acidic xylanase of bacillus acidocldarius 701 provided by the invention is 350-420IU/mL than vigor,, Applicable temperature scope is 40-75 ℃, 65 ℃ of optimal reactive temperatures, at 75 ℃ of enzymes complete stability alive; Applicable pH value in reaction scope is 5.0-11.0, in pH value, is 11.0 o'clock enzyme complete stabilities alive, and optimal reaction pH value is 10.0.
Press mutagenesis screening scheme, mutant strain step-sizing is eliminated, finally to strain excellent through leavening property test screen, obtain a strain and produce the bacterial strain bacillus acidocldarius 701 of high temperature resistant alkali body acidic xylanase, fermented liquid alkalescent xylanase is 350-420IU/mL than vigor, thermostability to enzyme is analyzed, and crude enzyme liquid is placed in respectively at 40 ℃, 45 ℃, 50 ℃, 55 ℃, 60 ℃, 65 ℃, 70 ℃, 75 ℃, every 10 minutes sampling and measuring enzymes, lives.At 40 ℃, 45 ℃, 50 ℃, 55 ℃, 60 minutes enzymes are lived and are not declined.At 60 ℃ and 65 ℃, what within 30 minutes, drop to constitutive enzyme work drops to 85% in 95%, 60 minute.At 70 ℃, what within 30 minutes, drop to constitutive enzyme work drops to 80% in 85%, 60 minute.At 75 ℃, what within 30 minutes, drop to constitutive enzyme work drops to 70% in 80%, 60 minute.
Another object of the present invention is the application of bacillus acidocldarius 701 in preparing high temperature resistant alkali body acidic xylanase.
Beneficial effect:
The prepared high temperature resistant alkali body acidic xylanase of bacillus acidocldarius 701 of the present invention is 350-420IU/mL than vigor,, Applicable temperature scope is 40-75 ℃, 65 ℃ of optimal reactive temperatures, at 75 ℃ of enzymes complete stability alive; Applicable pH value in reaction scope is 5.0-11.0, in pH value, is 11.0 o'clock enzyme complete stabilities alive, and optimal reaction pH value is 10.0.
Embodiment
Below by specific embodiment narration the present invention.Unless stated otherwise, in the present invention, technique means used is method known in those skilled in the art.In addition, embodiment is interpreted as illustrative, but not limits the scope of the invention, and the spirit and scope of the invention are only limited by claims.To those skilled in the art, do not deviating under the prerequisite of essence of the present invention and scope various changes that the material component in these embodiments and consumption are carried out or change and also belong to protection scope of the present invention.
Embodiment 1: the screening of starting strain
(1) from the waste paper pulp of Xue Li paper mill, Hunan Province, gather 2g soil sample, be added in the little triangular flask that contains the cooling sterilized water of 50mL, shaking table 200r/min, shake 20min, then be placed in 80 ℃ of water-baths, water-bath 10min, constantly shakes triangular flask and mixes soil sample, standing 5min draws supernatant liquor 100 μ L, successively gradient dilutions to 10
-1～10
-9concentration, chooses 10
-3, 10
-4, 10
-5, 10
-6concentration coating beef extract-peptone is dull and stereotyped, cultivates 24h for 37 ℃, continues 30 ℃ and cultivates 24h.
(2) according to the shape of microbe colony, size, surface tissue, marginal texture, quality, gloss, transparency, color and generation can lysochrome etc. the feature of aspect, with transfering loop, isolated single bacterium colony is chosen, move on on screening culture medium flat board, and numbering, 48h cultivated for 34 ℃.
Described screening culture medium consists of: extractum carnis 3g, sodium-chlor 5g, peptone 10g, agar powder 15g, distilled water l000mL, pH7.2,121 ℃ of sterilizing 20min.
(3) until bacterial strain, on flat board, cultivate after 48h, the dominant colony that difference is numbered on flat board is inoculated on screening culture medium inclined-plane, and preservation is used for multiple sieve.
(4) bacterial strain of the difference numbering of preservation is inoculated into respectively after slant activation 2 times, bacterial classification activate is accessed to seed culture medium, 180r/min, 37 ℃, cultivation 18h.By 5% inoculum size, be inoculated into fermention medium again, 200r/min, 28 ℃, fermentation 48h.By the centrifugal thalline that goes of gained fermented liquid 10000r/min, fermented liquid supernatant liquid carries out the experiment of alkalescent xylanase vitality test.
Described seed fermentation substratum consists of: glucose 2g, peptone 1g, NaH
2pO
42H
2o0.2g, Na
2hPO
42H
2o0.4g, MgSO
47H
2o0.05g, distilled water 100mL, pH7.4,121 ℃ of sterilizing 20min.
Described fermention medium consists of: glucose 2g, peptone 2g, NaH
2pO
42H
2o0.02g, Na
2hPO
42H
2o0.04g, MgSO
47H
2o0.01g, CaCl
20.01g, MnSO
40.002g, distilled water l00mL pH7.4,121 ℃ of sterilizing 20min.
(5) select the strong bacterial strain of product alkalescent xylanase enzyme activity and carry out glycerine preservation, obtain the starting strain HYX0021 of bacterial strain of the present invention.
The mutagenesis screening of embodiment 2 bacterial strains
(1) ultraviolet mutagenesis
Draw starting strain HYX0021 and cultivate 10-12h bacterium liquid, in injection plane ware, the bacterium liquid measure of each plane ware is 10mL.The plate that fills bacterium liquid is placed in mutagenesis case on the magnetic stirring apparatus under ultraviolet lamp tube, and plate is 30cm apart from the vertical range of ultraviolet lamp tube.Open ultraviolet lamp, preheating 30min, makes light wave stable.Regulate the rotating speed of stirrer, after stabilization of speed, open plate lid and irradiate, and start timing, the irradiation time of plate is respectively 1,2,3,4,5,6,7,8,9,10min.From each plate, take out the bacteria suspension of 0.1mL, do suitable dilution, obtain different dilution bacteria suspensions.Bacteria suspension 0.3mL coating isolation medium after absorption dilution is dull and stereotyped, is placed in thermostat container and cultivates (for avoiding photoreactivation plate to wrap up with black paper or kraft paper, whole mutagenesis operating process is all carried out under ruddiness).
(2) nitrosoguanidine mutagenesis
A little powder of nitrosoguanidine is placed in to plate culture medium central authorities, in culturing process, nitrosoguanidine progressively spreads, on flat board, forms the concentration gradient outside mediad, through cultivating the concentric(al) circles bacterium colony group can occur take that dull and stereotyped central authorities are the center of circle, dull and stereotyped central close region colony number is zero, and plate edge region bacterium colony increases successively.The bacterial classification that sets out produces the bacterial classification that makes a variation in various degree under different lethality rates.
(3) dull and stereotyped primary dcreening operation
Utilize transparent circle size to carry out primary dcreening operation.The bacterium colony dibbling growing after mutagenesis, on separating plate, is cultivated after 10-15h for 34-35 ℃, observe its transparent circle size around, choose bacterium colony that birch xylan transparent circle diameter and colony diameter ratio are larger and carry out shaking flask and sieve again.
Described separating plate substratum consists of: yeast powder 0.2%, birch xylan 0.2%, peptone 0.5%, extractum carnis 0.7%, dipotassium hydrogen phosphate 1.0%, sodium-chlor 0.1%, agar 1.8%, insufficient section pure water is supplied, pH7.0-8.0,121-123 ℃ of sterilizing 30-40min.
(4) shaking flask is sieved again
The colony inoculation that primary dcreening operation is obtained, in multiple sieve substratum, is cultivated 48 hours for 34 ℃, detects alkalescent xylanase enzyme and lives.
The described substratum that sieves again consists of: yeast powder 0.2%, and glucose 1%, peptone 0.4%, Semen Maydis powder 0.6%, dipotassium hydrogen phosphate 0.8%, insufficient section pure water is supplied, pH7.0-8.0,121-123 ℃ of sterilizing 30-40min.
(5) strain stability detects:
To sieve again the cultivation of going down to posterity of gained live high-enzyme strain, and follow the tracks of shaking flask and detect its enzyme and live.Through shaking flask, sieve again, finally determined a strain production bacterium, and by its called after 701, the characteristic of this bacterial strain is that product alkalescent xylanase efficiency is high, good stability.
The shaking flask result that superior strain 701 continuous passage is 6 times is as shown in table 1:
Table 1. bacterial strain 701 Detection of Stability results
| ? | Shaking flask enzyme activity (IU/mL) | Enzyme activity (%) |
| F1 | 380 | 100 |
| F2 | 382 | 100.6 |
| F3 | 388 | 102.0 |
| F4 | 378 | 99.5 |
| F5 | 388 | 102.3 |
| F6 | 387 | 101.9 |
Embodiment 3: identification of strains
Bacterial strain Physiology and biochemistry is identified: according to < < uncle Jie Shi Bacteria Identification handbook > > (the 9th edition) method, carry out the tests such as utilization of carbon source test, catalase test, hydrogen sulfide production test, indole test, nitrate and nitrite reduction test; Morphological Identification: dibbling is on beef-protein medium solid plate respectively by bacterial strain to be identified, and 37 ℃ of cultivation 2d, describe and record bacterium colony cultural characteristic and morphological features.Physiology and biochemistry identification mark is as shown in table 2.In table, result contrast < < uncle Jie Shi Bacteria Identification handbook > > belongs to feature with thermophilic bacillus species and conforms to.The extraction of molecular biology identification: bacterial genomes DNA is with reference to the method for < < molecular cloning experiment guide > > (the 3rd edition).According to the most conservative primers in bacterial 16 S rDNA.Primers F: 5'-AGA GTT TGA TCM TGGCTC AG-3'; Primer R:5'-AAG GAG GTG WTCCAR CC-3' is synthetic by Shanghai Sheng Gong biotechnology company limited.PCR reaction conditions: 94 ℃ of 5min, 94 ℃ of 30s, 55 ℃ of 40s, 72 ℃ of 2min, 30 circulations, 72 ℃ are extended 10min eventually.Pcr amplification product is delivered to the order-checking of Beijing San Bo polygala root Bioisystech Co., Ltd.Sequencing result carries out after two-way splicing, and the 16S rDNA sequence obtaining is submitted to and in NCBI nucleic acid database, carries out BLAST on-line analysis.From the GenBank nucleic acid database of NCBI, choose at random afterwards the part genus bacillus 16S rDNA sequence in report, application Clustal X 1.81 and Phylodraw082 software building systematic evolution tree are also analyzed.In conjunction with the physiology and morphology biochemical character of bacterial strain, preliminary evaluation fixes tentatively 701 bacterial strains into bacillus acidocldarius.
Table 2 bacterial strain 701 morphology and physio-biochemical characteristics
Note: result "+" represents positive; “ – " expression feminine gender.
Embodiment 4: the thermal stability analysis of alkalescent xylanase
Thermostability to enzyme is analyzed, and embodiment 2 crude enzyme liquids are placed in respectively at 40 ℃, 45 ℃, 50 ℃, 55 ℃, 60 ℃, 65 ℃, 70 ℃, every 10 minutes sampling and measuring enzymes, lives.At 40 ℃, 45 ℃, 50 ℃, 55 ℃ of crude enzyme liquids, 60 minutes enzymes are lived and are not declined.At 60 ℃ and 65 ℃, what within 30 minutes, drop to constitutive enzyme work drops to 85% in 95%, 60 minute.At 70 ℃, what within 30 minutes, drop to constitutive enzyme work drops to 80% in 85%, 60 minute.At 75 ℃, what within 30 minutes, drop to constitutive enzyme work drops to 70% in 80%, 60 minute, compared with prior art reaches same enzyme and lives, and tolerable temperature is higher.
Embodiment 5: alkalescent xylanase pH stability analysis
PH stability to enzyme is analyzed, and embodiment 2 crude enzyme liquids are placed in respectively to pH value 4.0,5.0,6.0,7.0,8.0,9.0,10.0,11.0 times, every 10 minutes sampling and measuring enzymes, lives.Crude enzyme liquid pH value 6.0,7.0,8.0,9.0 times, 60 minutes enzymes are lived and are not declined.PH value 5.0 and 10.0 times, what within 30 minutes, drop to that constitutive enzyme lives drops to 85% in 95%, 60 minute.PH value 4.0 and 11.0 times, what within 30 minutes, drop to that constitutive enzyme lives drops to 70% in 80%, 60 minute.It is obviously more wide in range than prior art bacterial strain that similarity condition is issued to same enzyme resistance to pH value alive.
Claims (2)
1. a bacillus acidocldarius (Bacillus thermophilus) 701 that produces high temperature resistant alkali body acidic xylanase, this bacterial strain is preserved in Chinese Typical Representative culture collection center on November 3rd, 2013, and preserving number is CCTCC NO:M2013537.
2. the application of bacillus acidocldarius as claimed in claim 1 (Bacillus thermophilus) 701 in preparing high temperature resistant alkali body acidic xylanase.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104031860A (en) * | 2014-05-27 | 2014-09-10 | 南京工业大学 | Bacillus sphaericus with high yield and high-temperature xylanase resistance and application thereof |
| CN105802880A (en) * | 2016-04-06 | 2016-07-27 | 安徽工程大学 | Alkalophilic bacillus and application thereof |
| CN111254094A (en) * | 2020-01-21 | 2020-06-09 | 深圳大学 | Bacillus alcalophilus, alkaline xylanase produced by bacillus alcalophilus and application of alkaline xylanase |
Citations (1)
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| CN101701205A (en) * | 2009-10-30 | 2010-05-05 | 中国农业科学院饲料研究所 | Alkali-resistant xylanase XynE2, genes thereof and application thereof |
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2013
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101701205A (en) * | 2009-10-30 | 2010-05-05 | 中国农业科学院饲料研究所 | Alkali-resistant xylanase XynE2, genes thereof and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| ALEXANDER KHASIN, ET AL: "Purification and Characterization of a Thermostable Xylanase from Bacillus stearothermophilus T-6", 《APPLIED AND ENVIRONMENTAL MICROBIOLOGY》 * |
| 解复红,等: "耐碱和耐热木聚糖酶研究进展", 《中国生物工程杂志》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104031860A (en) * | 2014-05-27 | 2014-09-10 | 南京工业大学 | Bacillus sphaericus with high yield and high-temperature xylanase resistance and application thereof |
| CN104031860B (en) * | 2014-05-27 | 2016-05-18 | 南京工业大学 | A kind of spherical bacillus of high-yield thermostable zytase and application thereof |
| CN105802880A (en) * | 2016-04-06 | 2016-07-27 | 安徽工程大学 | Alkalophilic bacillus and application thereof |
| CN105802880B (en) * | 2016-04-06 | 2019-07-30 | 安徽工程大学 | One plant of Bacillus alcalophilus and its application |
| CN111254094A (en) * | 2020-01-21 | 2020-06-09 | 深圳大学 | Bacillus alcalophilus, alkaline xylanase produced by bacillus alcalophilus and application of alkaline xylanase |
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