CN101880680A - Gene for coding beta-glucosaccharase and application thereof - Google Patents
Gene for coding beta-glucosaccharase and application thereof Download PDFInfo
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- CN101880680A CN101880680A CN2010101012116A CN201010101211A CN101880680A CN 101880680 A CN101880680 A CN 101880680A CN 2010101012116 A CN2010101012116 A CN 2010101012116A CN 201010101211 A CN201010101211 A CN 201010101211A CN 101880680 A CN101880680 A CN 101880680A
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- 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
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- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
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Abstract
The invention relates to a gene bgl for coding beta-glucosaccharase, which contains nucleotide sequence of SEQ ID NO.1 or functional equivalent variant. The invention comprises beta-glucosaccharase (SEQ ID NO.2) in relation to the gene or the functional equivalent variant thereof as well as application of the glucosaccharase in degrading cellulose.
Description
Technical field
The present invention relates to a kind of gene of encoding beta-glucosidase, particularly relate to the gene of clone from a kind of new encoding beta-glucosidase of anaerobism bacterial strain Clostridium.sp WGC702, the protein of this genes encoding can be used for cellulosic degraded.
Background technology
Cellulase is that a kind of multi-component enzyme is, this enzyme refers to energy degraded cellulose β-1, the general name of the class of enzymes of 4-glucoside bond, therefore, cellulase has the title of cellulase mixture again, is that a complicated enzyme of being made up of multiple lytic enzyme is, mainly from fungi and bacterium (Gao Fengju, Li Chunxiang, the progress of fungi and bacteria cellulose enzyme, the pedagogical journal in Tangshan).In the major part of bacterium belongs to, all there is bacterial strain to degrade and utilizes Mierocrystalline cellulose, they can be divided into three monoids from physiology characteristic: anaerobic fermentation type, normally gram-positive microorganism, aerobic type gram-positive microorganism, aerobic sliding-type bacterial strain (Lynd L R, Weimer P J, ZylW.H.V, et al.Microbial cellulose utilization:fundamentals andbiotechnology.Microbiology and Molecular Biology Reviews, 2002,66 (3) .506-557).The cellulase of natural cellulose of degrading all is complicated multi-enzyme system, high-efficiency decomposition of cellulose needs the synergy of three kinds of enzymes: 1. endoglucanase (endo-1,4-β-D-glucanase, EC3.2.1.4, be called for short EG) act on the noncrystalline domain of cellulosic molecule inside, hydrolysis β-1 immediately, 4-glycosidic link, with the cellulosic molecule brachymemma, produce the small molecules Mierocrystalline cellulose of a large amount of band non reducing ends; 2. circumscribed dextran cellobiohydrolase (exo-1,4-β-D-glucanase, EC3.2.1.91) claim cellobiohydrolase (cellobiohydrolase again, be called for short CBH), it contains two kinds of components: CBH I and CBH II act on Mierocrystalline cellulose linear molecule end, hydrolysis β-1, the 4-glycosidic link is cut next cellobiose molecule at every turn; 3. beta-glucosidase (β-1,4-glucosidase, EC3.2.1.21, be called for short BG), this fermentoid is hydrolyzed into glucose molecule (Tomme P, Warrenraj with cellobiose, Gilkes N R.Cellulosehydrolysis by bacteria and fungi.Adv Microbiol Physiol,, 1995,37.1-81).
Three kinds of cellulases of aerobic bacteria be with separately independently form to be secreted into cell external cellulolytic, three kinds of Mierocrystalline celluloses of anaerobic bacterium are then united a kind of fiber corpusculum of formation, be combined on the cell walls Mierocrystalline cellulose (the Lynd L R that degrades, Weimer P J, Zyl W.H.V, et al.Microbial cellulose utilization:fundamentals and biotechnology.Microbiology and Molecular Biology Reviews, 2002,66 (3) .506-557).
Beta-glucosidase is a kind of in the cellulase system, is the rate-limiting enzyme of cellulose degradation.Beta-glucosidase belongs to hydrolase, extensively is present in the natural various organism, and from the bacterium to the plant, animal all contains this enzyme, and the different function of travelling therein.In microorganism (yeast, aspergillus tubigensis, trichoderma and bacterium) cell, it mainly is used as a kind of of cellulase, hydrolysis is incorporated into the β-D-glycosidic link of end, irreducibility, discharge β-D-glucose and corresponding aglucon (Wang Huafu, You Xiaoqing simultaneously, the mensuration of activity of beta-glucosidase [J] in the tealeaves. Chinese tea, 1996 (3) .16-17).In cellulosic saccharification, the beta-glucosidase function is that cellobiose and Mierocrystalline cellulose oligosaccharides are hydrolyzed into glucose.The hydrolysis of beta-glucosidase and saccharification can both be used in industrial production.But in cellulosic component, beta-glucosidase content is few, vigor is low, becomes the bottleneck (Shao Jinhui of cellulose degradation, Han Jinxiang, Zhu Youming, Wu's screen, Wu Jianmei, beta-glucosidase cure the application in field workers and peasants. biological chemistry, 2005,25 (1) .22-24).BGL is in cellulase preparation simultaneously, and protein content is minimum, only accounts for about 1%.
Main application of beta-glucosidase is exactly to be used for cellulose degradation.Mierocrystalline cellulose is that glucose is with 1,4 glycosidic link bonded polymkeric substance, be the constituent of cell walls, account for the 1/2-1/3 of plant dry weight, the Mierocrystalline cellulose that the whole world was produced by photosynthesis in a year reaches 1,000 hundred million tons, be the abundantest renewable resources (Pan Lihua, Luo Jianping, the research of beta-glucosidase and application progress, Food science, 2006,27 (12) .803-806).If Mierocrystalline cellulose can be applied to the fermented food industry raw material, can make us break away from absolute dependence to cereal grain, alleviate world energy sources (Xu Jing in short supply,, Zhang Yongzhong, Sun Yanmei, the progress of beta-glucosidase, food research and development, 2005,26 (6) .183-186).Make up engineering bacteria by gene recombination technology, secreting, expressing high reactivity beta-glucosidase has great importance to cellulosic effective degraded.
Most to the research report of beta-glucosidase at present the aerobic fungi, mainly be the mould aspect.In recent years, though there is research the bacterium aspect, study considerably less.Reason has: one is relevant with the position of cellulase in cell of Production by Bacteria.Two is relevant with the secretory volume of bacterium extracellular enzyme.Simultaneously because the cellulase that bacterium the produced overwhelming majority is extracellular enzyme, vigor is also lower, therefore, directly extracting cellulase from bacterium secretory product is feasible (Pierre Beguin et al.Bacterial celluase.Biochemical society Transaction, 1992 (20) .42-46) substantially.But, increase (Gao Fengju, Li Chunxiang, fungi and bacteria cellulose enzyme progress, the pedagogical journal in Tangshan, 2005,27 (2) .7-10) day by day with the feasibility of bacterium cellulase-producing in next life along with the development of genetic engineering technique.From otherwise application such as environment protection and anaerobically fermentings, be very important to the research of bacterium especially anerobe and facultative anaerobe.Along with research, will further widen the cellulosic scope of utilizing to anerobe and facultative anaerobe cellulase-producing.Therefore, from bacterium, especially extracting cellulase from anerobe and facultative anaerobe will and become an important directions extracting cellulase with its high efficiency.
Summary of the invention
The present invention relates to a kind of gene bgl (SEQ ID NO.1) of new encoding beta-glucosidase, it is to separate the genomic library of the strain anaerobism bacterial strain Clostridium.sp WGC702 that preserves from this chamber to obtain.A positive colony foreign DNA in this bacterial strain library (SEQ ID NO.1) is by 1344bp based composition, open reading frame (the Open Reading Frame that contains complete beta-glucosidase gene bgl, ORF), the initiation codon of bgl gene is ATG, and terminator codon is TAG.
The protein of SEQ ID NO.2 is the beta-glucosidase product B GL of gene bgl coding, form by 448 amino acid, 2-446 amino acids from the N end is family's 1 glycosyl hydrolase (glycosylhydrolase) functional domain, the catalysis territory (e-value be 0.0) for the hypothetical protein of clostridium (Clostridium.celerecrescens AJ295659) the highest with bgl catalysis territory homology, both similaritys are 84%.
Gene bgl can degraded cellulose at the recombinant products BGL of expression in escherichia coli.
The invention still further relates to and contain expression carrier of the present invention, and be used to transform the host of gene of the present invention.
The invention provides a kind of new beta-glucosidase gene, the beta-glucosidase of this coded by said gene has purposes widely in cellulosic degraded.
Embodiment
Following implementation method is for better explanation the present invention, and the purpose that should not be construed as limiting the invention.
Used in an embodiment of the present invention material comprises: intestinal bacteria (Escherichia coli) strain is Tuner (DE3), DH5 α and expression vector pET-blue2 (belonging to the component available from the protein expression and the purification kit of Novagen company); Cloning vector is the cloning vector pGEM3zf (+) available from Promega company; Reclaim test kit available from the DNA purifying of Quigen company, available from reagent such as the restriction enzyme of Promega, MBI, dephosphorylation enzymes.
Below by embodiment the present invention is described in detail:
1) genomic library of structure Clostridium.sp WGC702
Extract total DNA of Clostridium.sp WGC702, use different restriction endonuclease (BamHI, HindIIII respectively, EcoRI, PstI) enzyme is cut total DNA, and the endonuclease reaction system is: enzyme 10u, total DNA 50ng, buffer 2uL, sterilized water mend to cumulative volume be 20uL.The enzyme tangent condition is reaction 3h under 37 ℃ of water-baths, after reacting completely, get the effect that its enzyme of 10uL electrophoresis detection is cut, selection can not be only enzyme for the enzyme of small segment too (most of for 3kb following) with dna fragmentation digestion, selects suitable enzyme to cut system again and enzyme is cut the time.DNA product size after discovery is cut with the BamHI enzyme at last is most of about 4-10Kb, so select the BamHI enzyme as making up the library enzyme.
Cut total DNA with the incomplete enzyme of BamHI, the enzyme system of cutting is: enzyme 20U; Total DNA 1ug; Buffer 30uL; Last water is added to cumulative volume 300uL, and reaction system is reacted 1h under 37 ℃ of water-baths, and electrophoresis, glue reclaim the endonuclease bamhi of 2-10Kb as the foreign DNA that makes up the library.
The BamHI enzyme is cut pGEM-3zf (+) carrier, and dephosphorylation is handled again, and treatment condition are: the carrier DNA of 0.5ug, and the dephosphorylation enzyme of adding 10U, buffer is 5uL, adding water to final volume is 50uL, reaction 1h 30min in 37 ℃ of water-baths.After having reacted, product carries out purifying (removing the impurity and the enzyme of the inside) with pure-cycle kit PCR purification column, again according to carrier: the system of external source=1: 4～1: 10 is done ligation, contain carrier DNA 100ng in the reaction system, foreign DNA 17ng, T4DNA ligase enzyme 0.1～1U, adding water, to make final volume be 10 μ l, connects down in 16 ℃ and spend the night.After will connecting product purification, getting 10uL is transferred among the E.coli DH5 α by electricity, coat X-gal and IPTG and contain and carry out overnight incubation in the white 37 ℃ of thermostat containers of screen plate of indigo plant on the LA flat board of acillin, picking white recon point Vitamin C2 screen plate then, in 37 ℃ of thermostat containers after the overnight incubation, picking can generate the clone of black hydrolysis circle on the Vitamin C2 plate, the present invention only relates to one of them clone, called after p-bgl, further this clone is inoculated in the liquid LB substratum, extracts plasmid after 37 ℃ of shaking table overnight incubation.Cut the plasmid of this positive colony with the BamHI enzyme, the electrophoresis detection enzyme is cut the result, and after cutting through the BamHI enzyme, what obtain is to be about 3.5Kb BamHI external source fragment.
2) mensuration of beta-glucosidase bgl sequence and sequential analysis
P-bgl is delivered Shanghai give birth to worker bio-engineering corporation mensuration dna nucleotide sequence.Software on the sequence usefulness NCBI (National Center for BiotechnologyInformation, http://www.ncbi.nlm.nih.gov) that records such as ORF finder (
Http:// www.ncbi.nlm.nih.gov/gorf/gorf.html),
Blast (
Http:// www.ncbi.nlm.nih.gov/BLAST) dna sequence dna is analyzed.The open reading frame of gene bgl (Open Reading Frame, ORF) by 1344bp based composition, contain complete beta-glucosidase gene bgl open reading frame (Open Reading Frame, ORF), the initiation codon of bgl gene is ATG, and terminator codon is TAG.
3) amino acid sequence analysis of the product B GL of beta-glucosidase gene bgl coding
One of beta-glucosidase gene bgl coding contains 448 amino acid whose protein, is 53978.6 dalton with this proteinic theoretical molecular size of DNAStar software prediction.
With simple assemblies structural research instrument (Simple Modular Architecture Research Tool, SMART, http://smart.embl-heidelberg.de) unit construction of analysis BGL, the result is that this aminoacid sequence does not have the clear signal peptide, and the 2-446 amino acids of holding from N is family's 1 glycosyl hydrolase (glycosyl hydrolase) functional domain.
4) clone of beta-glucosidase gene bgl and expression
Use upstream primer 5 '-CAGGATCCGATGGGACAGAGACAGTTTC-3 ' downstream primer 5 '-CGAAGCTTGTATACCGCATTCTGCTC-3 ', by polymerase chain reaction (PCR) amplification beta-glucosidase gene bgl, reaction conditions is 95 ℃ of 4min, 95 ℃ of 40sec; 55 ℃ of 55sec; 72 ℃ of 2min30sec; 30 circulations altogether, 72 ℃ of 5min30sec.After cutting beta-glucosidase gene bgl with restriction enzyme BamHI and HindIII enzyme, and be connected with the expression vector pET-blue2 that the HindIII enzyme is cut through BamHI.To connect product again and pass through CaCl
2Method is transformed among the intestinal bacteria Tuner (DE3), is applied to screening and cloning on the LA flat board that contains penbritin (100 μ g/mL).Further extract this clone's plasmid DNA and with its called after pET-bgl, cut pET-bgl with restriction enzyme HindIII and BamHI enzyme after, carry out 0.7% agarose gel electrophoresis analysis, pET-bgl contains a DNA external source fragment about 1.3kb as a result.
The recombination bacillus coli Tuner (DE3) that will contain plasmid pET-bgl is inoculated into 20mL and contains in the LB substratum of penbritin (100 μ g/mL), and 37 ℃ of shaking culture are treated OD
600Be 0.4 o'clock, it is 0.5mmol/L that adding IPTG makes its final concentration, induces 4 hours for 37 ℃.11000 rev/mins centrifugal 3 minutes, collect thalline, with the resuspended thalline of phosphoric acid buffer of 4mL pH7.0100mmol/L, the broken born of the same parents of ultrasonic wave 9 minutes.12000 rev/mins centrifugal 10 minutes, supernatant is the beta-glucosidase BGL crude enzyme liquid that contains.
5) beta-glucosidase BGL enzyme mensuration alive
1. drawing 0.2ml enzyme liquid, to join 2ml pH value be 4.8 0.2molL
-1Na
2HPO
4-0.1molL
-1In the citrate buffer solution, place 50 ℃ of water bath heat preservation 10min simultaneously with 2ml 5mM PNPG solution;
2. said two devices is mixed, 50 ℃ of water-baths accurately are incubated 10min;
3. the Na that adds 2ml 1M
2CO
3Stop enzyme reaction, room temperature leaves standstill 5min;
4. calculate production concentration according to the p-NP typical curve.
The enzyme activity unit definition: the enzyme activity that every min enzyme liquid hydrolysis PNPG generates 1 μ mol p-NP is the enzyme U of unit alive.
Sequence table
＜110〉Guangxi University
＜120〉a kind of gene of encoding beta-glucosidase and application thereof
<160>2
<170>PatentIn?Version?3.3
<210>1
<211>1344
<212>DNA
＜213〉fusobacterium (Clostridium.sp)
<220>
<221>gene
<222>(1)...(1344)
<223>
<400>1
atgggacaga?gacagtttcc?ggaaggcttt?gcatgggggg?cggctactgc?gtcataccag 60
attgaaggcg?cttgggatga?agacggaaga?ggagaaacga?tatgggacag?gtattgccag 120
gttcccggta?atgttttaaa?tggagatgat?ggaaaaatag?catgtgacca?ttatcacaga 180
tacaaagagg?atgtagctct?gatgaagcgt?atgggaatca?aggcttaccg?gttctccatt 240
gcctggtcca?ggatttttcc?aaagggatat?ggggaagtca?atgaaaaggg?gcttgaattt 300
tacagccgtc?ttattgatga?acttctggat?gcgggaatcg?aaccgtatat?cacgttatac 360
cactgggatc?ttcctcaggc?cctgcaggat?ataggcggat?gggccaatcc?ccagatgccg 420
gaatattttt?tggaatacgc?aaaggttgtt?atagatgcat?ttcatggcaa?ggtgaagaaa 480
tggatcacat?taaatgaacc?ttattgcgct?gcgtttctag?gcaattccga?ggggcgtcag 540
gctcccggta?tccgggattt?ttcgacagca?gtgcaggtgt?catactatct?ttatgtagga 600
cacggtcttg?cagtggatta?tttccgtaaa?aaaggctgtg?acggggaaat?tggaattacc 660
ttaaacctga?tgggaaggct?tcctcttacg?ggtaagccag?aggatataca?ggcagcagcc 720
agggcagatg?ggtatttaaa?ccgctggttt?gctgagccta?ttgtgtacgg?cagatatcca 780
caggaaatgt?tggagtttta?ccgttccaaa?ggtgtaagac?ttccggaatt?taaagaggag 840
cacatgaagc?tcattggaca?gaagctggat?ttcatcggcc?tgaattatta?caatgacttt 900
catgtgaagg?cggatgacag?cgtatggcct?cttgggttta?agatagagaa?tcccaaatac 960
gttcccgtca?atgacaggaa?ctggccggta?acagagcaag?gcctggtaac?gatgctttta 1020
cggatgaaga?atgagtatgg?aatagacaag?atctacataa?cggaaaatgg?cacttccttc 1080
ccggatgtgg?tttccatgga?aggaaaagta?gaagacggcg?caaggaagga?ttatctccac 1140
aggcatttaa?cagcactttg?ggaggccatt?tcccagggag?ttaatgttca?gggatatttt 1200
cagtggtctt?tgtatgacaa?ttttgagtgg?tcattcggat?atagcagccg?tttcggaatc 1260
gtattcgtgg?acttcaatac?ccaggaaaga?attattaagg?aaagcggaca?ctggtattca 1320
aatgtaattg?agcagaatgc?ggta 1344
<210>2
<211>448
<212>PRT
＜213〉fusobacterium (Clostridium.sp)
<400>2
Met?Gly?Gln?Arg?Gln?Phe?Pro?Glu?Gly?Phe?Ala?Trp?Gly?Ala?Ala
1 5 10 15
Thr?Ala?Ser?Tyr?Gln?Ile?Glu?Gly?Ala?Trp?Asp?Glu?Asp?Gly?Arg
16 20 25 30
Gly?Glu?Thr?Ile?Trp?Asp?Arg?Tyr?Cys?Gln?Val?Pro?Gly?Asn?Val
31 35 40 45
Leu?Asn?Gly?Asp?Asp?Gly?Lys?Ile?Ala?Cys?Asp?His?Tyr?His?Arg
46 50 55 60
Tyr?Lys?Glu?Asp?Val?Ala?Leu?Met?Lys?Arg?Met?Gly?Ile?Lys?Ala
61 65 70 75
Tyr?Arg?Phe?Ser?Ile?Ala?Trp?Ser?Arg?Ile?Phe?Pro?Lys?Gly?Tyr
76 80 85 90
Gly?Glu?Val?Asn?Glu?Lys?Gly?Leu?Glu?Phe?Tyr?Ser?Arg?Leu?Ile
91 95 100 105
Asp?Glu?Leu?Leu?Asp?Ala?Gly?Ile?Glu?Pro?Tyr?Ile?Thr?Leu?Tyr
106 110 115 120
His?Trp?Asp?Leu?Pro?Gln?Ala?Leu?Gln?Asp?Ile?Gly?Gly?Trp?Ala
121 125 130 135
Asn?Pro?Gln?Met?Pro?Glu?Tyr?Phe?Leu?Glu?Tyr?Ala?Lys?Val?Val
136 140 145 150
Ile?Asp?Ala?Phe?His?Gly?Lys?Val?Lys?Lys?Trp?Ile?Thr?Leu?Asn
151 155 160 165
Glu?Pro?Tyr?Cys?Ala?Ala?Phe?Leu?Gly?Asn?Ser?Glu?Gly?Arg?Gln
166 170 175 180
Ala?Pro?Gly?Ile?Arg?Asp?Phe?Ser?Thr?Ala?Val?Gln?Val?Ser?Tyr
181 185 190 195
Tyr?Leu?Tyr?Val?Gly?His?Gly?Leu?Ala?Val?Asp?Tyr?Phe?Arg?Lys
196 200 205 210
Lys?Gly?Cys?Asp?Gly?Glu?Ile?Gly?Ile?Thr?Leu?Asn?Leu?Met?Gly
211 215 220 225
Arg?Leu?Pro?Leu?Thr?Gly?Lys?Pro?Glu?Asp?Ile?Gln?Ala?Ala?Ala
226 230 235 240
Arg?Ala?Asp?Gly?Tyr?Leu?Asn?Arg?Trp?Phe?Ala?Glu?Pro?Ile?Val
241 245 250 255
Tyr?Gly?Arg?Tyr?Pro?Gln?Glu?Met?Leu?Glu?Phe?Tyr?Arg?Ser?Lys
256 260 265 270
Gly?Val?Arg?Leu?Pro?Glu?Phe?Lys?Glu?Glu?His?Met?Lys?Leu?Ile
271 275 280 285
Gly?Gln?Lys?Leu?Asp?Phe?Ile?Gly?Leu?Asn?Tyr?Tyr?Asn?Asp?Phe
286 290 295 300
His?Val?Lys?Ala?Asp?Asp?Ser?Val?Trp?Pro?Leu?Gly?Phe?Lys?Ile
301 305 310 315
Glu?Asn?Pro?Lys?Tyr?Val?Pro?Val?Asn?Asp?Arg?Asn?Trp?Pro?Val
316 320 325 330
Thr?Glu?Gln?Gly?Leu?Val?Thr?Met?Leu?Leu?Arg?Met?Lys?Asn?Glu
331 335 340 345
Tyr?Gly?Ile?Asp?Lys?Ile?Tyr?Ile?Thr?Glu?Asn?Gly?Thr?Ser?Phe
346 350 355 360
Pro?Asp?Val?Val?Ser?Met?Glu?Gly?Lys?Val?Glu?Asp?Gly?Ala?Arg
361 365 370 375
Lys?Asp?Tyr?Leu?His?Arg?His?Leu?Thr?Ala?Leu?Trp?Glu?Ala?Ile
376 380 385 390
Ser?Gln?Gly?Val?Asn?Val?Gln?Gly?Tyr?Phe?Gln?Trp?Ser?Leu?Tyr
391 395 400 405
Asp?Asn?Phe?Glu?Trp?Ser?Phe?Gly?Tyr?Ser?Ser?Arg?Phe?Gly?Ile
406 410 415 420
Val?Phe?Val?Asp?Phe?Asn?Thr?Gln?Glu?Arg?Ile?Ile?Lys?Glu?Ser
421 425 430 435
Gly?His?Trp?Tyr?Ser?Asn?Val?Ile?Glu?Gln?Asn?Ala?Val
436 440 445 448
Claims (6)
1. the gene of an encoding beta-glucosidase is characterized in that, has SEQ ID NO.1 nucleotide sequence or its functional equivalent varient.
2. the gene of claim 1, wherein said functional equivalent varient is the mutant form of the nucleotide sequence of SEQ ID NO.1, mutant form comprises disappearance, nonsense, insertion, missense.
3. according to the protein of the coded by said gene of claim 1, its aminoacid sequence is shown in SEQ ID NO.2.
4. an expression vector is characterized in that, it contains the described gene of claim 1.
5. a host cell is characterized in that, it contains prokaryotic cell prokaryocyte or the eukaryotic cell that the described expression vector of claim 3 transforms.
6. the application during the described protein of claim 3 is handled at cellulose degradation with to cellulose-containing material.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010101012116A CN101880680A (en) | 2010-01-26 | 2010-01-26 | Gene for coding beta-glucosaccharase and application thereof |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102268421A (en) * | 2011-08-03 | 2011-12-07 | 山西恩泽生物技术有限公司 | Cloning, expression and application of beta-glucosaccharase gene |
| CN102517307A (en) * | 2011-12-29 | 2012-06-27 | 山东大学 | Beta-glucosidase Mut1b as well as expressed gene and application thereof |
| CN103525844A (en) * | 2013-09-27 | 2014-01-22 | 广西大学 | Gene bglG for coding beta-glucosaccharase and application of gene bglG |
-
2010
- 2010-01-26 CN CN2010101012116A patent/CN101880680A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102268421A (en) * | 2011-08-03 | 2011-12-07 | 山西恩泽生物技术有限公司 | Cloning, expression and application of beta-glucosaccharase gene |
| CN102517307A (en) * | 2011-12-29 | 2012-06-27 | 山东大学 | Beta-glucosidase Mut1b as well as expressed gene and application thereof |
| CN102517307B (en) * | 2011-12-29 | 2013-01-30 | 山东大学 | Beta-glucosidase Mut1b as well as expressed gene and application thereof |
| CN103525844A (en) * | 2013-09-27 | 2014-01-22 | 广西大学 | Gene bglG for coding beta-glucosaccharase and application of gene bglG |
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