CN111073877A - Xylanase with excellent temperature stability and pH tolerance and application thereof - Google Patents

Xylanase with excellent temperature stability and pH tolerance and application thereof Download PDF

Info

Publication number
CN111073877A
CN111073877A CN201911280407.3A CN201911280407A CN111073877A CN 111073877 A CN111073877 A CN 111073877A CN 201911280407 A CN201911280407 A CN 201911280407A CN 111073877 A CN111073877 A CN 111073877A
Authority
CN
China
Prior art keywords
xylanase
enzyme
activity
leu
glu
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
Application number
CN201911280407.3A
Other languages
Chinese (zh)
Inventor
信丰学
吕阳
姜岷
蒋羽佳
董维亮
章文明
方艳
马江锋
周杰
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Nanjing Tech University
Original Assignee
Nanjing Tech University
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Nanjing Tech University filed Critical Nanjing Tech University
Publication of CN111073877A publication Critical patent/CN111073877A/en
Pending legal-status Critical Current

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
    • C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
    • C12N9/2477—Hemicellulases not provided in a preceding group
    • C12N9/248—Xylanases
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
    • C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
    • C12P7/16—Butanols
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
    • C12P7/44—Polycarboxylic acids
    • C12P7/46—Dicarboxylic acids having four or less carbon atoms, e.g. fumaric acid, maleic acid
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
    • Y02E50/00—Technologies for the production of fuel of non-fossil origin
    • Y02E50/10—Biofuels, e.g. bio-diesel

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Zoology (AREA)
  • Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Enzymes And Modification Thereof (AREA)

Abstract

The invention discloses xylanase with excellent temperature stability and pH tolerance and application thereof. The enzyme can keep activity for more than 9 hours without reduction in a pH6.5 buffer system at the temperature of between 35 and 65 ℃. The xylanase can maintain activity in buffer solution with pH of 4.0-10.0 at 4 deg.c for over 12 hr, and may be obtained from anaerobic thermophilic bacteria and used in decomposing xylan at different temperature and pH. Can be used in the process of producing butanol by fermenting clostridium acetobutylicum at 37 ℃, enlarges the substrate universality, enhances the utilization of xylan and reduces the production cost.

Description

Xylanase with excellent temperature stability and pH tolerance and application thereof
Technical Field
The invention belongs to the technical field of biology, and particularly relates to xylanase with excellent temperature stability and pH tolerance.
Background
Xylan is an abundant renewable resource in nature, is most representative hemicellulose, occupies 1/3-1/2 of the hemicellulose, and is the most abundant polysaccharide in nature except cellulose. Compared with cellulose, hemicellulose is easier to degrade and convert by microorganisms.
The xylanase is the most key hydrolase in a xylanase hydrolase system, and can hydrolyze xylan into low xylans such as xylooligosaccharide and xylobiose and a small amount of xylose and arabinose by hydrolyzing a xylose molecule β -1, 4-glycosidic bond, so that the xylan can be degraded by treating corncobs with the xylanase, and energy is provided for microorganisms taking xylose as a carbon source.
Disclosure of Invention
The invention aims to provide xylanase with excellent temperature stability and pH tolerance and application thereof.
In order to achieve the aim, the invention separates and purifies xylanase from a thermophilic anaerobic bacterium.
A xylanase with excellent temperature stability and pH tolerance has an amino acid sequence shown in SEQ ID NO. 1.
The DNA sequence of the coding gene of the xylanase is shown as SEQ ID NO. 2.
The xylanase can keep activity for more than 9 hours at the temperature of 35-65 ℃ without reduction, has excellent temperature stability, can keep activity for more than 12 hours in a buffer solution with the pH value of 4.0-10.0, and has excellent pH tolerance.
The enzyme is derived from a strainThermoanaerobacterium thermosaccharolyticmM5。
An expression vector comprising said xylanase gene.
A recombinant bacterium obtained by transforming a host cell with the expression vector.
The xylanase can keep the activity for at least 9 hours without reduction at the pH of 6.5 and between 35 ℃ and 65 ℃; the activity of the enzyme can be maintained in a buffer solution with pH of 4.0-10.0 at 4 ℃ for more than 12 hours without reduction.
The application of the xylanase in degrading xylan.
The xylanase is applied to the application industries of producing biological butanol, brewing, feed, papermaking, food spinning and the like.
The expression vector and the expression system containing the xylanase coding gene.
Has the advantages that:
the xylanase provided by the invention has excellent properties, and is suitable for application industries of biological butanol, brewing, feed, papermaking, food spinning and the like. The xylanase of the invention has excellent temperature stability and pH tolerance. The enzyme can keep activity for more than 9 hours without reduction in a buffer system with pH6.5 and between 35 ℃ and 65 ℃. The activity of the enzyme can be maintained in a buffer solution with pH of 4.0-10.0 at 4 ℃ for more than 12 hours without reduction.
Drawings
FIG. 1 is an SDS-PAGE electrophoresis of xylanase;
FIG. 2 shows the variation of xylanase activity with temperature;
FIG. 3 shows the variation of xylanase activity with pH;
FIG. 4 is a graph of the change in xylanase activity thermostability;
FIG. 5 is a graph showing the change in pH tolerance of xylanase activity.
The patent requires that the application numbers filed in 2019, 5 months and 20 days are as follows: 201910417176X.
Detailed Description
The present invention will be further described with reference to the following specific examples.
The experimental procedures used in the following examples are all conventional procedures unless otherwise specified.
Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
Example 1 preparation and purification of xylanases
The xylanase is derived from bacterial strainThermoanaerobacterium thermosaccharolyticmM5, deposited in China center for type culture Collection with the date of preservation being 2017, 2 months and 27 days and the preservation number being CCTCC NO: M2017072, obtaining the xylanase sequence in the strain through a colony pcr, removing a terminator, connecting the xylanase sequence into pET29a (+) or pET28a (+) after removing the terminator, introducing the connected plasmid into Escherichia coli DH5 α, and then introducing the correctly verified plasmid into Escherichia coli BL21 (DE 3) for subsequent induced expression and purification of xylanase.
Composition of LB medium: 5g/L yeast extract (yeast extract), 5g/L NaCl and 10g/L peptone (peptone).
Shaking to OD at 37 deg.C600After the concentration is 0.6-0.8, adding IPTG (isopropyl thiogalactoside) with the final concentration of 0.5 mM, inducing for 16-24 hours at 16-37 ℃, recording all liquid in a shake flask as fermentation liquid, centrifuging the fermentation liquid at 6000 rpm for 10min at 4 ℃, collecting thalli, suspending and centrifuging the obtained thalli containing the target protein twice by using phosphate buffer solution (0.05mM PBS (pH6.5)) and adding 20ml of heavy suspension bacterial sludge for the third time, fully suspending each thalli cell and then crushing the cell by adopting an ultrasonic method. Centrifuging the obtained cell disruption solution at 4 deg.C and 8000rpm for 10min, and filtering the obtained supernatant with 0.22 μm filter membrane to obtain crude enzyme solution of thallus.
The protein purification is carried out by the following specific method: after ethanol elution from the Ni-NTA-Sefiniose column, the column was equilibrated by adding 5 column volumes of sterile water, phosphate buffer (50mM pH6.5 PBS). The crude enzyme solution was added, followed by sequentially adding elution buffers (50mM NaH2PO4, 300mM NaCl) having imidazole concentrations of 50mM, 100mM, 200mM, 250mM, 350mM, pH8.0, respectively, to elute, and collecting eluates in stages. The eluates of xylanase 200mM and 250mM imidazole were each replaced with PBS buffer (PBS buffer, 50mM, pH 6.5) using a 10kDa ultrafiltration tube to remove imidazole in the enzyme solution, thereby obtaining a xylanase enzyme solution. The protein solution obtained by the above purification was subjected to SDS-PAGE, and the purification results are shown in FIG. 1, wherein 1 is the purified enzyme solution, 2 is the crude enzyme solution, and the obtained xylanase has a molecular weight of 47.7 kDa.
Example 2 enzymatic characterization
The xylanase obtained by the method is subjected to the following reactions for enzyme characterization:
1. determination of enzymatic Activity of xylanase
Using the purified xylanase as described above, the following reactions were carried out: mu.l of the xylanase solution obtained above at a concentration of 0.646. mu.g/ml was diluted 5-fold and 100. mu.l was added to 100. mu.l of 1% xylan solution, respectively, wherein the xylan was dissolved in phosphate buffer (50mM pH6.5 PBS), and reacted at 55 ℃ for 10 minutes. The amount of reducing sugar produced after completion of the reaction was measured by the 3.5-dinitrosalicylic acid (DNS) method.
After the reaction was completed, 200. mu.l of DNS solution was added, boiled for 5 minutes, cooled, added with 2.1ml of water and mixed well, and the absorbance at 540nm was measured with a spectrophotometer.
One unit of enzyme activity is defined as the amount of enzyme required to hydrolyze a substrate to produce 1. mu. mol reducing sugars per minute.
Specific enzyme activity is defined as the ratio of enzyme activity to the amount of the corresponding protein. The enzyme activity reaction system is shown in table 1.
TABLE 1 enzyme activity reaction System
Components Blank space Experimental control group Sample set Sample control group
Buffer solution 200μl 100μl 100μl
Substrate 100μl 100μl
Enzyme solution 100μl 100μl
2. Determination of optimum reaction temperature and optimum reaction pH of xylanase
The xylanase purified in example 1 was subjected to enzymatic reaction in different pH ranges (50mM acetic acid-sodium acetate buffer at pH4.0, 4.5, 5.0, 5.5, 6.0; PBS buffer at pH 6.0, 6.5, 7.0, 7.5, 8.0; glycine-sodium hydroxide buffer at pH8.0, 8.5, 9.0, 9.5, 10.0) to determine the optimum reaction pH. The results show that the xylanase has an optimum pH of 6.5, and the results are shown in FIG. 3. The xylanase has enzyme activity in different temperature ranges (35, 40, 45, 50, 55, 60, 65, 70, 75 and 80 ℃) measured under the condition of the optimum reaction pH value, and the optimum reaction temperature is determined to be 65 ℃, and the result is shown in figure 2.
3. Enzyme temperature stability and pH tolerance
The pure enzyme solution of example 1 was diluted 5-fold with 50mM PBS buffer solution having a pH of 6.5, and the enzyme activity was measured using the diluted enzyme solution. The diluted enzyme solution was recorded as a diluted enzyme solution.
The diluted enzyme solution was placed in a water bath at 35 ℃, 40 ℃, 45 ℃, 50 ℃, 55 ℃, 60 ℃, 65 ℃, 70 ℃, 75 ℃ and 80 ℃ for 10, 20, 40 and 60 minutes, respectively, to determine the residual activity of the enzyme. The pH value in the enzyme activity determination reaction system is 6.5, the determination temperature is 55 ℃, and the rest conditions and steps are the same as those in experiment 1. The experiment was repeated 3 times. The diluted enzyme solution was left in an incubator at 35 ℃ for 9 hours, and the residual activity of the enzyme was measured every hour, except for the same conditions as above.
The results are shown in figure 4, and show that the enzyme activity is not reduced after the treatment for 60 minutes at the temperature of 35-65 ℃; the enzyme is shown to have good temperature stability. When the enzyme is placed in an incubator at 35 ℃ for 9 hours, the activity is not reduced, which shows that the enzyme is suitable for decomposing corn cobs at 37 ℃ and stably utilizes wastes.
The purified enzyme solution (prepared in example 1) was left at 4 ℃ for 1h and 12h under different pH conditions (extensive buffer pH 4.0-10.0), and then subjected to enzymatic reaction at 55 ℃ and pH7.0, with untreated enzyme solution as a control. Reacting for 10min by using xylan as a substrate to determine the enzyme activity. After being treated by the buffer solution with the pH value of 4.0-10.0 for 1 hour and 12 hours, the enzyme activity is not reduced. The results are shown in FIG. 5, which shows that the pH tolerance is excellent, and the activity can be kept for a long time under different pH conditions. Therefore, the method has good application prospect in industrial application.
Example 3 use of xylanases for the fermentative production of butanol
Clostridium acetobutylicum LJ4 (strain preservation number is CCTCC NO: M2017754) takes 30g/L xylan as a substrate, the fermentation temperature is 35-39 ℃, the fermentation pH is 5.0-5.5, the rotating speed is 0-120rpm, the xylanase inoculation amount is 2.5% v/v, the strain inoculation amount is 2.5% v/v, the fermentation period is 5-7 days, and the concentration of the product butanol is 4.6 g/L.
Example 4 application of xylanase in fermentative production of succinic acid
Actinobacillus succinogenes (strain number is 130Z), 30g/L xylan is used as a substrate, the fermentation temperature is 35-39 ℃, the fermentation pH is 6.0-7.5, the rotation speed is 100-200 rpm, the xylanase inoculation amount is 2.5% v/v, the strain inoculation amount is 5% v/v, the fermentation period is 48h, and the concentration of succinic acid produced is 30.5 g/L.
Sequence listing
<110> Nanjing university of industry
<120> xylanase with excellent temperature stability and pH tolerance and application thereof
<140>201910417176X
<141>2019-05-20
<160>2
<170>SIPOSequenceListing 1.0
<210>1
<211>413
<212>PRT
<213> xylanase (2 Ambystoma latex x Ambystoma jeffersonanum)
<400>1
Met Asn Ala Asp Ala Ala Asp Lys Leu Lys His Arg Lys Gly Ile Ala
1 5 10 15
Lys Ile Lys Leu Val Lys Lys Asp Gly Ser Pro Ile Lys Asp Ala Glu
20 25 30
Val Ala Val Ser Gln Val Lys His Lys Phe Leu Phe Gly Cys Gly Ala
35 40 45
Phe Asp Ser Leu Pro Leu Ala Asn Gly Glu Leu Lys Glu Asn Asp Lys
50 55 60
Glu Lys Ile Glu Asp Arg Phe Glu Lys Phe Phe Asp Leu Phe Asn Tyr
65 70 75 80
Ala Thr Ile Pro Phe Tyr Trp Gly Arg Phe Glu Pro Glu Lys Gly Lys
85 90 95
Pro Asp Thr Asn Arg Leu Lys Lys Ala Ser Glu Trp Leu Val Ser Lys
100 105 110
Gly Cys Leu Val Lys Gly His Pro Leu Cys Trp His Thr Val Thr Ala
115 120 125
Pro Trp Leu Leu Asp Met Asn Asn Glu Asp Ile Leu Lys Ala Gln Leu
130 135 140
Ser Arg Ile Lys Arg Glu Val Ser Asp Phe Lys Gly Leu Val Asn Ile
145 150 155 160
Trp Asp Val Ile Asn Glu Val Val Ile Met Pro Ile Phe Asp Lys Tyr
165 170 175
Asp Asn Gly Ile Thr Arg Ile Cys Lys Glu Leu Gly Arg Ile Arg Leu
180 185 190
Val Lys Glu Val Phe Asn Glu Ala Lys Lys Ala Asn Pro Glu Ala Val
195 200 205
Leu Leu Ile Asn Asp Phe Asn Thr Ser Ile Ser Tyr Glu Ile Leu Ile
210 215 220
Glu Gly Cys Leu Glu Ala Gly Ile Pro Ile Asp Ala Ile Gly Ile Gln
225 230 235 240
Ser His Met His Gln Gly Tyr Trp Gly Val Glu Lys Thr Leu Glu Val
245 250 255
Leu Glu Arg Phe Ser His Phe Asn Ile Pro Leu His Phe Thr Glu Asn
260 265 270
Thr Leu Leu Ser Gly His Leu Met Pro Pro Glu Ile Glu Asp Leu Asn
275 280 285
Asp Tyr Gln Ile Arg Asp Trp Pro Ser Thr Pro Asp Gly Glu Glu Arg
290 295 300
Gln Ala Met Glu Ile Val Gln His Tyr Lys Thr Leu Phe Ser His Pro
305 310 315 320
Met Val Glu Ser Ile Thr Trp Trp Asn Phe Cys Asp Glu Asn Ala Trp
325 330 335
Leu Gly Ala Pro Ala Gly Leu Leu Arg Arg Asp Asn Ser Cys Lys Pro
340 345 350
Ser Tyr Tyr Glu Leu Lys Lys Leu Ile Lys Asp Glu Trp Trp Thr His
355 360 365
Pro Thr Arg Leu Val Thr Ser Asn Thr Gly Glu Phe Glu Phe Thr Gly
370 375 380
PheLeu Gly Glu Tyr Glu Leu Val Ile Ser Asp Lys Arg Phe Tyr Phe
385 390 395 400
Thr Leu Asp Lys Asn Ser Thr Thr Ile Glu Ile Thr Ile
405 410
<210>2
<211>1242
<212>DNA
<213> xylanase (2 Ambystoma latex x Ambystoma jeffersonanum)
<400>2
atgaatgcgg atgctgcaga taaattaaaa caccgcaaag gaatcgccaa aataaagtta 60
gttaagaaag atggttcacc tataaaagat gcagaagtcg ctgtatctca ggtgaagcat 120
aaatttttat ttggttgtgg agcatttgat tctcttcctc ttgccaatgg tgaattgaaa 180
gaaaatgata aagaaaaaat tgaagaccgt tttgagaaat tttttgactt atttaactat 240
gctacgattc cattttattg gggcaggttc gagcctgaaa aaggaaagcc agacacaaat 300
agactcaaaa aggcttcaga atggcttgta tcaaaaggtt gccttgtaaa aggccatcca 360
ctttgctggc atactgtaac agcaccttgg cttttagata tgaacaatga agacatatta 420
aaggctcagc tatcccgcat aaaacgtgaa gtaagcgatt ttaaaggatt agtgaatata 480
tgggatgtaa taaatgaagt tgtcatcatg cctatttttg ataaatacga caatggaata 540
accaggatat gcaaagaatt aggccgtatt cgcctcgtaa aagaagtttt taatgaagct 600
aaaaaagcta atcctgaagc agttctcctt ataaatgact ttaatacatc aatttcatac 660
gaaatactca tagaaggatg ccttgaagct ggaatcccta ttgatgccat aggaattcag 720
tcacacatgc atcaaggata ttggggagtt gaaaaaactt tagaagtact tgaaagattt 780
tcgcatttca acattccatt gcattttaca gaaaacacat tattatcagg gcatttgatg 840
ccacctgaaa tagaagactt aaatgactac cagataaggg attggccttc aacacctgat 900
ggtgaagaac gacaagcgat ggaaatagta cagcattaca aaactctttt ttcacatcca 960
atggtagaat caattacgtg gtggaatttt tgtgatgaaa atgcatggct tggtgcacca 1020
gcaggtcttt tacgccgtga caattcatgc aaaccatcat attacgaatt aaaaaagctt 1080
ataaaagatg aatggtggac acatcctaca cgtcttgtca caagtaacac aggcgaattt 1140
gaatttacag gtttcttagg cgaatacgaa ttagtcatta gcgataaaag gttttacttt 1200
acccttgata aaaacagcac aacaattgaa atcacgattt aa 1242

Claims (8)

1. A xylanase with excellent temperature stability and pH tolerance, which is characterized in that the amino acid sequence is shown as SEQ ID NO. 1.
2. The gene of the xylanase of claim 1, wherein the DNA sequence of the gene is shown in SEQ ID NO. 2.
3. The xylanase according to claim 1, characterized in that the enzyme can keep activity for more than 9 hours between 35 ℃ and 65 ℃ without reduction, has excellent temperature stability, can keep activity for more than 12 hours in buffer solution with pH4.0-10.0, and has excellent pH tolerance.
4. The xylanase according to claim 1, characterized in that the enzyme is derived fromFrom bacterial strainsThermoanaerobacterium thermosaccharolyticmM5。
5. An expression vector comprising the xylanase gene of claim 2.
6. A recombinant bacterium obtained by transforming a host cell with the expression vector according to claim 5.
7. The xylanase according to claim 3, characterized in that the xylanase retains activity for at least 9 hours at a pH of 6.5, between 35 ℃ and 65 ℃; the activity of the enzyme can be maintained in a buffer solution with pH of 4.0-10.0 at 4 ℃ for more than 12 hours without reduction.
8. Use of a xylanase according to claim 1 for degrading xylan.
CN201911280407.3A 2019-05-20 2019-12-13 Xylanase with excellent temperature stability and pH tolerance and application thereof Pending CN111073877A (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201910417176X 2019-05-20
CN201910417176 2019-05-20

Publications (1)

Publication Number Publication Date
CN111073877A true CN111073877A (en) 2020-04-28

Family

ID=70314458

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201911280407.3A Pending CN111073877A (en) 2019-05-20 2019-12-13 Xylanase with excellent temperature stability and pH tolerance and application thereof

Country Status (1)

Country Link
CN (1) CN111073877A (en)

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104726480A (en) * 2013-12-18 2015-06-24 中国科学院过程工程研究所 High temperature xylanase gene and high temperature alpha-glucuronidase gene, and protein expression methods and applications thereof
CN106754827A (en) * 2016-11-16 2017-05-31 南京林业大学 A kind of thermostable xylanase and preparation method and application
CN106995790A (en) * 2017-06-02 2017-08-01 南京工业大学 Bacterial strain for directly producing butanol by using xylan as unique carbon source and application thereof
CN107760753A (en) * 2017-12-07 2018-03-06 南京工业大学 Method for producing butanol by co-culture fermentation of high-temperature anaerobe for pyrolyzing sugar and clostridium acetobutylicum
CN108330091A (en) * 2018-03-30 2018-07-27 南京工业大学 Clostridium acetobutylicum and application thereof
CN109536565A (en) * 2018-12-20 2019-03-29 南京工业大学 Method for producing succinic acid by utilizing mixed fermentation of high-temperature anaerobic bacteria for pyrolyzing sugar and actinobacillus succinogenes

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104726480A (en) * 2013-12-18 2015-06-24 中国科学院过程工程研究所 High temperature xylanase gene and high temperature alpha-glucuronidase gene, and protein expression methods and applications thereof
CN106754827A (en) * 2016-11-16 2017-05-31 南京林业大学 A kind of thermostable xylanase and preparation method and application
CN106995790A (en) * 2017-06-02 2017-08-01 南京工业大学 Bacterial strain for directly producing butanol by using xylan as unique carbon source and application thereof
CN107760753A (en) * 2017-12-07 2018-03-06 南京工业大学 Method for producing butanol by co-culture fermentation of high-temperature anaerobe for pyrolyzing sugar and clostridium acetobutylicum
CN108330091A (en) * 2018-03-30 2018-07-27 南京工业大学 Clostridium acetobutylicum and application thereof
CN109536565A (en) * 2018-12-20 2019-03-29 南京工业大学 Method for producing succinic acid by utilizing mixed fermentation of high-temperature anaerobic bacteria for pyrolyzing sugar and actinobacillus succinogenes

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
NCBI: "Accession No：WP_094043097.1", Retrieved from the Internet <URL:GenBank> *
NCBI: "WP_094043097.1" *
YUJIA JIANG ET AL.: "The Draft Genome Sequence of Thermophilic Thermoanaerobacterium thermosaccharolyticum M5 Capable of Directly Producing Butanol from Hemicellulose", pages 620 - 623 *
李琦 等: "耐热木聚糖酶基因的克隆表达及其在酶解玉米芯木聚糖中的应用", pages 63 - 69 *

Similar Documents

Publication Publication Date Title
CN112481240B (en) GH16 family heat-resistant glucanase mutant and construction method and application thereof
CN109628433B (en) Pullulanase with high secretion capacity and application thereof
JP2019503688A (en) Lactic acid-utilizing bacteria genetically modified to secrete polysaccharide degrading enzymes
CN109385413B (en) Glucoamylase TlGA1931 and gene and application thereof
JP4998991B2 (en) Method for producing cellulase
US20200347371A1 (en) Variants of exoglucanases having improved actvity and uses thereof
CN112592914A (en) Special green alga polysaccharide lyase and production process thereof
CN116064484B (en) Mutant chitinase SsChi18A-2 and application thereof
ES2885684T3 (en) Exoglucanase variants with enhanced activity and their uses
CN105154417A (en) Acidic cellulose from fungus and gene and application thereof
CN111073877A (en) Xylanase with excellent temperature stability and pH tolerance and application thereof
CN106978410B (en) Bifunctional glucanase with chitosan hydrolysis activity, gene, vector, engineering bacterium and application thereof
WO2020057476A1 (en) Application of trehalase in fermentative production
WO2021153587A1 (en) Filamentous fungus trichoderma mutant strain
CN102676476B (en) Dextranase with improved enzyme activity and thermal stability
JPWO2019188980A1 (en) Trichoderma Risei mutant strain and protein production method
US10000780B2 (en) Endoglucanase variants having improved activity, and uses of same
CN116064485B (en) Mutant chitinase SsChi18A-3 and application thereof
US20150218593A1 (en) Polypetide with Reinforced Beta-Glucosidase Activity at Low Temperature
JP6518107B2 (en) Transcription factor mutant
CN117925577B (en) Method for improving xylanase activity, xylanase and application thereof
CN118222549B (en) High-temperature-resistant xylanase mutant and application thereof
CN114807095B (en) Chitinase mutant and application thereof
CN118240804B (en) Xylanase mutant and application thereof
JP5777128B2 (en) Novel cellulase

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
RJ01 Rejection of invention patent application after publication
RJ01 Rejection of invention patent application after publication

Application publication date: 20200428