CN105754884A - Strain capable of efficiently expressing alkaline pectinase and application of strain - Google Patents

Strain capable of efficiently expressing alkaline pectinase and application of strain Download PDF

Info

Publication number
CN105754884A
CN105754884A CN201610170070.0A CN201610170070A CN105754884A CN 105754884 A CN105754884 A CN 105754884A CN 201610170070 A CN201610170070 A CN 201610170070A CN 105754884 A CN105754884 A CN 105754884A
Authority
CN
China
Prior art keywords
alkaline
strain
genetic engineering
engineering bacterium
alkaline pectinase
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
CN201610170070.0A
Other languages
Chinese (zh)
Inventor
刘松
陈双全
陈坚
堵国成
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.)
Jiangnan University
Original Assignee
Jiangnan 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 Jiangnan University filed Critical Jiangnan University
Priority to CN201610170070.0A priority Critical patent/CN105754884A/en
Publication of CN105754884A publication Critical patent/CN105754884A/en
Pending legal-status Critical Current

Links

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
    • C12N9/88—Lyases (4.)
    • 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
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/80—Vectors or expression systems specially adapted for eukaryotic hosts for fungi
    • C12N15/81—Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y402/00—Carbon-oxygen lyases (4.2)
    • C12Y402/02—Carbon-oxygen lyases (4.2) acting on polysaccharides (4.2.2)
    • C12Y402/02002—Pectate lyase (4.2.2.2)
    • D—TEXTILES; PAPER
    • D06—TREATMENT OF TEXTILES OR THE LIKE; LAUNDERING; FLEXIBLE MATERIALS NOT OTHERWISE PROVIDED FOR
    • D06M—TREATMENT, NOT PROVIDED FOR ELSEWHERE IN CLASS D06, OF FIBRES, THREADS, YARNS, FABRICS, FEATHERS OR FIBROUS GOODS MADE FROM SUCH MATERIALS
    • D06M16/00—Biochemical treatment of fibres, threads, yarns, fabrics, or fibrous goods made from such materials, e.g. enzymatic
    • D—TEXTILES; PAPER
    • D21—PAPER-MAKING; PRODUCTION OF CELLULOSE
    • D21C—PRODUCTION OF CELLULOSE BY REMOVING NON-CELLULOSE SUBSTANCES FROM CELLULOSE-CONTAINING MATERIALS; REGENERATION OF PULPING LIQUORS; APPARATUS THEREFOR
    • D21C5/00—Other processes for obtaining cellulose, e.g. cooking cotton linters ; Processes characterised by the choice of cellulose-containing starting materials
    • D21C5/005—Treatment of cellulose-containing material with microorganisms or enzymes
    • 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
    • C12N2800/00—Nucleic acids vectors
    • C12N2800/10—Plasmid DNA
    • C12N2800/102—Plasmid DNA for yeast
    • 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
    • C12N2800/00—Nucleic acids vectors
    • C12N2800/22—Vectors comprising a coding region that has been codon optimised for expression in a respective host

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biochemistry (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Biomedical Technology (AREA)
  • General Health & Medical Sciences (AREA)
  • Biotechnology (AREA)
  • Molecular Biology (AREA)
  • Mycology (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Plant Pathology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Textile Engineering (AREA)
  • Medicinal Chemistry (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Enzymes And Modification Thereof (AREA)

Abstract

The invention discloses a strain capable of efficiently expressing alkaline pectinase and an application of the strain and belongs to the technical field of gene engineering. Alkaline pectinase genes are expressed in Pichia pastoris GS115 with a gene recombination technology, and the strain GS115-PIC9K-PGL is obtained and has a remarkably improved yield by comparison with the yield before sequence optimization. According to the strain, the enzyme activity of the alkaline pectinase fermented for 72 h in a shaking flask is 301.9 U/ml and is improved by about 25%, and a good basis is laid for large-scale production of the alkaline pectinase. The alkaline pectinase produced with the strain can catalyze alpha-1,4 glycosidic bonds of polygalacturonic acid to be pyrolysed through trans-elimination under the alkaline condition and is widely applied to food, textile, papermaking and other industries.

Description

The bacterial strain of a kind of high efficient expression alkaline pectase and application thereof
Technical field
The present invention relates to bacterial strain and the application thereof of a kind of high efficient expression alkaline pectase, belong to gene engineering technology field.
Background technology
Pectase is a kind of compound enzyme, it is possible to Pectin polymers resolves into unsaturated oligogalacturonans.This enzyme is widely distributed, It is found in section parasitic nematicide, plant and microorganism.Pectase is widely used, the commercial Application history of existing more than 40 year. Pectase is divided into acid pectase and alkaline pectase PGL by the difference according to optimal reaction pH.Wherein acid pectase is main It is applied to the aspects such as clear juice fruit wine, extraction fruit and vegetable juice, fruit decortication.PGL application is mainly used in weaving, food, makes Paper industry and environmental area.Enzyme process effect above-mentioned field correlated response has environmental protection, economize in raw materials consumptive material and reaction condition temperature in application With etc. advantage.But it is less that PGL carries out molecular modification research at present, the PGL carrying out commercialization is the most little.
The host being presently expressed by alkaline pectase is mainly Pichia sp., bacillus subtilis and escherichia coli.Although by fermentation The means such as optimization can be effectively improved the yield of alkaline pectase, but when the yield reaching certain limit alkaline pectase can not enter one Step improves, and limits the industrialized production of alkaline pectase, therefore need to solve to limit the factor of alkaline pectin expression of enzymes from source.
Although Pichia sp. host has expressing protein is prone to the advantages such as purification, but current alkaline pectase is in Pichia sp. During expression, have that expression is the highest or exogenous gene original nucleotide sequences is not very suitable for asking of Pichia sp. host Topic, thus limit the alkaline pectase high efficient expression in Pichia sp..
Therefore, it is necessary to improve alkaline pectase further to produce the production capacity of bacterial strain more to adapt to industrialized needs.
Summary of the invention
In order to solve the problems referred to above, the invention provides the alkaline pectase of a kind of optimization, and express finishing of this alkaline pectase Red yeast gene engineering bacteria.
Described alkaline pectinase gene nucleotide sequence sequence as shown in SEQ ID NO.1.
Described pichia yeast genetic engineering bacteria, is nucleotide sequence alkaline pectinase gene as shown in SEQ ID NO.1 to be connected On Expression vector pPIC9K, then it is transformed into and Pichia sp. host strain obtains.
In one embodiment of the invention, described expression vector is pPIC9K.
In one embodiment of the invention, described Pichia sp. host is Pichiapastoris GS115.
The present invention also provides for the construction method of a kind of described pichia yeast genetic engineering bacteria, and step is as follows:
(1) use chemistry complete synthesis codon optimized after alkaline pectinase gene PGL；
(2) alkaline pectinase gene that step (1) obtains is connected on yeast expression vector pPIC9K, is recombinated Plasmid pPIC9K-PGL；
(3) it is excellent that recombiant plasmid pPIC9K-PGL conversion Pichia pastoris GS115 step (2) obtained obtains codon The engineering strain expressing alkaline pectase changed.
The present invention also provides for a kind of method utilizing described engineering bacteria fermentation to produce alkaline pectase, is to be lived by genetic engineering bacterium Be seeded to after change in growth medium BMGY in 30 DEG C, cultivate 24h under the conditions of 220rpm, transfer the most again into inducing culture In base BMMY, in 23 DEG C, shake flask fermentation, the expression of inducible alkaline pectase under the conditions of 220rpm.
In one embodiment of the invention, described activation is to be inoculated in seed culture medium YPD cultivation 14h；Again with 5% Inoculum concentration proceed to growth medium BMGY.
In one embodiment of the invention, during described induction, every 24h adds the methanol of 1.5% (v/v).
The present invention is also claimed the application in terms of food, weaving or papermaking of the described genetic engineering bacterium.
Beneficial effects of the present invention:
For the original engineering strain before gene optimization, the engineering strain Pichia pastoris of the present invention GS115-pPIC9K-PGL enzyme is lived and is improve 25%, and the enzyme in shaking flask level is lived and be can reach 301.9U/ml (72h), is existing Top level in the Pichia sp. having been reported that, higher enzyme work can be realized by tank fermentation and medium optimization on fermentation tank, Large-scale production for alkaline pectase is had laid a good foundation.The alkaline pectase that the present invention obtains can be urged in the basic conditions Change and cracked by the α of trans-elimination polygalacturonic acid-Isosorbide-5-Nitrae glycosidic bond, be widely used in the industry such as food, textile and paper.
Figure of description
Fig. 1: clonal expression plasmid map；
Fig. 2: alkaline pectase SDS-PAGE analyzes；1-3 swimming lane represents GS115-pPIC9K, GS115-pPIC9K-PGL respectively After optimization, before GS115-pPIC9K-PGL optimization；
The performance comparison of the alkaline pectase that strain fermentation produces before and after the optimization of Fig. 3: alkaline pectinase gene.
Detailed description of the invention:
Culture medium:
Seed culture medium YPD (1L): tryptone 20g, yeast powder 10g, glucose 20g.
Growth medium BMGY (1L): peptone 20g, yeast powder 10g, glycerol 40g, YNB 13.4g, pH6.0's The phosphate buffer of 0.1M.
Inducing culture BMMY (1L): peptone 20g, yeast powder 10g, methanol 9%, YNB 13.4g, pH6.0's The phosphate buffer of 0.1M.
Alkaline pectase enzyme activity determination:
Use spectrophotometry.Unit enzyme is lived and is defined: unit interval cracking polygalacturonic acid produces the unsaturation of 1 μm ol Enzyme amount used by polygalacturonic acid.Enzyme activity determination condition is: enzyme activity detects: fermentation liquid 8000rpm is centrifuged 10min, born of the same parents Outer PGL is i.e. contained among fermented supernatant fluid, takes a certain amount of detection.PGL reaction system: containing 0.2% polygalacturonic acid Glycine-NaOH buffer (the 0.2mol L of (substrate)-1, 0.44mmol L-1CaCl2, pH9.4) 2mL, to be measured Sample 20 μ L, inactive enzyme liquid is blank.PGL reaction condition is: reaction system is placed in water-bath 15min at 45 DEG C, With 3mL phosphoric acid solution (0.03mol L-1) terminate reaction, at 235nm, measure absorbance.
Embodiment 1: the structure of recombinant bacterium and qualification
Alkaline pectinase gene PGL of chemistry complete synthesis sequence such as SEQ ID NO.1, is redesigned primer, is obtained by the method for PCR Alkaline pectinase gene PGL that must optimize, is cloned on Expression vector pPIC9K, it is thus achieved that recombiant plasmid pPIC9K-PGL (clonal expression plasmid map is shown in accompanying drawing 1), converts Pichiapastoris GS115 by recombinant vector, obtains restructuring through Screening and Identification Bacterial strain Pichiapastoris GS115-pPIC9K-PGL.
Primer is following (sequence is respectively as shown in SEQ ID NO.2 and SEQ ID NO.3):
PGL upstream: GCTGAAGCTTACGTAGAATTCGCTGATTTGGGTCATCAAACACTTG
PGL downstream: AAGGCGAATTAATTCGCGGCCGCTTAGTTCAATTTTCCAGCACCTGCT
The conversion of Pichia sp. uses electrotransformation.
Specifically comprise the following steps that single colony inoculation of picking yeast recipient bacterium in 25mLYPD fluid medium, 30 DEG C of shaking table mistakes Night；With 5% inoculum concentration switching 50mL YPD fluid medium, 30 DEG C of shaking tables are cultivated to OD600=1.3-1.5；4 DEG C centrifugal 5000 Rpm, 5min, abandon supernatant；With 50mL ice pre-cooling sterilized water, thalline is resuspended；4 DEG C of centrifugal 5000rpm, 5min, abandon supernatant； With 25mL ice pre-cooling sterilized water, thalline is resuspended；4 DEG C of centrifugal 5000rpm, 5min, abandon supernatant；Again with 5mL 1mol L-1's The sorbitol washes of ice pre-cooling 1 time, resuspended, 4 DEG C, 5000rpm is centrifuged 5min, abandons supernatant；Add appropriate volume 1mol L-1 The sorbitol of ice pre-cooling, resuspended；In subpackage extremely aseptic EP pipe, often pipe 80ul, in case converting.The coexpression vector that will extract PPIC9K-PGL enzyme Sal I linearisation, adds linearizing plasmid 1-5 μ g in 80 μ l competent yeast cells and places on ice 15 minutes, being rapidly added in 0.2cm electric shock cup (ice pre-cooling), 1500v shocks by electricity, is rapidly added the sorbitol of 1mL ice pre-cooling, Being coated with MD flat board, after cultivating 3-4 days, picking monoclonal carries out next step screening.
Embodiment 2: the alkaline pectase enzyme activity determination of engineering strain and protein electrophoresis
Cultural method: bacterial strain is inoculated into basic fermentation medium YPD after seed activation, in 30 DEG C, cultivate under the conditions of 220rpm 14h, be forwarded to the growth medium BMGY after optimizing cultivate based on 30 DEG C, cultivate 24h under the conditions of 220rpm, then by bacterial strain Proceeding to 23 DEG C, 220rpm in inducing culture BMMY, every 24h adds the methanol of 1.5%, the expression of inducible alkaline pectase.
Selecting green skies PAGE gel electrophoresis kit to prepare 12% separation gel and 5% and concentrate glue, concrete operation method is shown in product Product description.Sample and 5 × sample-loading buffer mix with volume ratio 4:1, boiling water bath 10min, loading after cooling.During electrophoresis, 80V Constant-voltage, after to be instructed dose enters separation gel, voltage is adjusted to 150V, and to be instructed dose to terminating electrophoresis time at the bottom of glue.Use coomassie Gel is dyeed by light blue dyeing liquor, dyeing 1h rear decoloring (accompanying drawing 2 is shown in by SDS-PAGE collection of illustrative plates).
Determine the alkaline pectase enzyme in the fermentation liquid that above-mentioned shake-flask culture method obtains to live simultaneously.Result shows, expresses SEQ The recombinant bacterium Pichiapastoris GS115-pPIC9K-PGL of the alkaline pectinase gene of ID NO.1, the enzyme work in shaking flask is 301.9U/ml (72h), than the recombinant bacterium enzyme of the alkaline pectinase gene (sequence is as shown in SEQ ID NO.4) expressed before optimizing Live 242.6U/ml (72h) improve about 25% and Biomass be basically unchanged (Fig. 3).The present invention is the big of alkaline pectase Large-scale production is had laid a good foundation.
Additionally, inventor has also tested the alkaline pectinase gene after other multiple sequence optimisation, with Pichiapastoris GS115, PPIC9K is vector construction recombinant bacterium, it was found that expression effect is not as the present invention, such as expresses the base of SEQ ID NO.5 The recombinant yeast pichia pastoris of cause, alkaline pectase enzyme under the same conditions is lived and is only 253.2U/ml (72h), is far smaller than this Bright.
Embodiment 3: the purification of alkaline pectase
Recombinant bacterium fermentation liquid 8000r/min is centrifuged 20min, takes supernatant, add ammonium sulfate and carry out gradient and saltout, low-temperature centrifugation Collection 30～50% ammonium sulfate precipitation part, be dissolved in the enzyme of salt precipitation in Glycine-NaOH buffer solution (pH7.5), By 20mmol/L Glycine-NaOH buffer solution dialysis treatment 24h.Centrifugal gained supernatant enters through cation-exchange chromatography Row is the most isolated and purified.

Claims (8)

1. a pichia yeast genetic engineering bacteria, it is characterised in that described genetic engineering bacterium expresses nucleotide sequence such as SEQ ID Alkaline pectinase gene shown in NO.1.
Genetic engineering bacterium the most according to claim 1, it is characterised in that described genetic engineering bacterium is with Pichia pastoris GS115 is that host builds and obtains.
Genetic engineering bacterium the most according to claim 1, it is characterised in that described genetic engineering bacterium is with pPIC9K as table Reach carrier, Pichia pastoris GS115 is host expresses alkaline pectinase gene.
4. the method for a fermentation production of alkaline pectic enzyme, it is characterised in that be to utilize the genetic engineering bacterium described in claim 1.
Method the most according to claim 4, it is characterised in that described method is raw by being seeded to after genetic engineering bacterium activation In long culture medium BMGY in 30 DEG C, cultivate 24h under the conditions of 220rpm, transfer in inducing culture BMMY the most again, In 23 DEG C, the expression of inducible alkaline pectase under 220rpm.
Method the most according to claim 5, it is characterised in that in Induction Process, every 24h adds the methanol of 1.5%.
7. the genetic engineering bacterium of claim 1 produces the alkaline pectase obtained.
8. the alkaline pectase of claim 7 application in terms of food, weaving, environment or papermaking.
CN201610170070.0A 2016-03-23 2016-03-23 Strain capable of efficiently expressing alkaline pectinase and application of strain Pending CN105754884A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201610170070.0A CN105754884A (en) 2016-03-23 2016-03-23 Strain capable of efficiently expressing alkaline pectinase and application of strain

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201610170070.0A CN105754884A (en) 2016-03-23 2016-03-23 Strain capable of efficiently expressing alkaline pectinase and application of strain

Publications (1)

Publication Number Publication Date
CN105754884A true CN105754884A (en) 2016-07-13

Family

ID=56345636

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201610170070.0A Pending CN105754884A (en) 2016-03-23 2016-03-23 Strain capable of efficiently expressing alkaline pectinase and application of strain

Country Status (1)

Country Link
CN (1) CN105754884A (en)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106399286A (en) * 2016-10-18 2017-02-15 江南大学 Method for promoting Pichia pastoris to produce alkaline pectinase
CN106399337A (en) * 2016-10-18 2017-02-15 江南大学 Alkaline pectinase producing recombinant bacteria and application thereof
CN106520587A (en) * 2016-10-18 2017-03-22 江南大学 Recombinant strain producing alkaline polygalacturonate lyase and application thereof

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101157900A (en) * 2007-09-04 2008-04-09 江南大学 Alkaline pectic enzyme producing engineering strain and its construction and method for producing alkaline pectic enzyme with the same
CN102559638A (en) * 2010-12-17 2012-07-11 武汉新华扬生物股份有限公司 Alkaline pectinase poly lactic acid (PLA) and gene and application thereof
CN102604977A (en) * 2011-11-22 2012-07-25 湖北大学 Optimized nucleotide sequence of alkaline pectinase pell68s and high-level expression method thereof
CN103966195A (en) * 2014-05-07 2014-08-06 江南大学 Alkaline pectinase mutant with improved specific and enzyme activity

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101157900A (en) * 2007-09-04 2008-04-09 江南大学 Alkaline pectic enzyme producing engineering strain and its construction and method for producing alkaline pectic enzyme with the same
CN102559638A (en) * 2010-12-17 2012-07-11 武汉新华扬生物股份有限公司 Alkaline pectinase poly lactic acid (PLA) and gene and application thereof
CN102604977A (en) * 2011-11-22 2012-07-25 湖北大学 Optimized nucleotide sequence of alkaline pectinase pell68s and high-level expression method thereof
CN103966195A (en) * 2014-05-07 2014-08-06 江南大学 Alkaline pectinase mutant with improved specific and enzyme activity

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
张成杰: "Bacillus subtilis 168碱性果胶酶基因在毕赤酵母中的优化、表达、特性以及应用的研究", 《中国优秀硕士学位论文全文数据库 基础科学辑》 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106399286A (en) * 2016-10-18 2017-02-15 江南大学 Method for promoting Pichia pastoris to produce alkaline pectinase
CN106399337A (en) * 2016-10-18 2017-02-15 江南大学 Alkaline pectinase producing recombinant bacteria and application thereof
CN106520587A (en) * 2016-10-18 2017-03-22 江南大学 Recombinant strain producing alkaline polygalacturonate lyase and application thereof
CN106399286B (en) * 2016-10-18 2019-12-24 江南大学 Method for promoting pichia pastoris to produce alkaline pectinase

Similar Documents

Publication Publication Date Title
AU2011214324A1 (en) Host cell capable of producing enzymes useful for degradation of lignocellulosic material
CN104130951A (en) Recombinant pichia pastoris engineering bacteria and metabolic recombinant xylanase as well as preparation of metabolic recombinant xylanase
Kadowaki et al. Xylanase production by Aspergillus tamarii
CN107142225A (en) A kind of pichia yeast recombinant bacterium of overexpression Streptomyces sp. FA1 sources zytase
CN105754884A (en) Strain capable of efficiently expressing alkaline pectinase and application of strain
CN105018448A (en) Heat-resisting acidic cellulase of fungus source and gene and application thereof
CN105886484A (en) Thermophilic cellulase, encoding gene thereof and application of thermophilic cellulase
CN105838728B (en) Polygalacturonase optimization gene and its expression vector and application
CN105802867A (en) Alkaline pectinase secretion-enhanced strain and application thereof
CN105316310A (en) Alkaline pectinase mutant with improved specific enzyme activity and heat stability
CN107384899A (en) The acid protease g412 and its gene of a kind of originated from fungus and application
CN105950491A (en) Strain for efficiently expressing alkaline pectate and construction and application thereof
CN105713850A (en) Pichia pastoris strain capable of constitutively expressing alkaline pectinase and application thereof
CN110373403A (en) High temperature resistant neutrality Pullulanase and its application
CN106047840B (en) A kind of acidity exo polygalacturonase and its gene and application
CN106520587B (en) It is a kind of produce alkaline pectase recombinant bacterium and its application
CN109694859A (en) A kind of thermophilic pectase and its expressing gene and application
CN106884030A (en) The method for improving the fermentation yield of the fusion protein of containing cellulose binding domain
CN103966195A (en) Alkaline pectinase mutant with improved specific and enzyme activity
CN102363772B (en) Acidic cellulase EGI, gene thereof and application thereof
CN111893107A (en) Pichia pastoris engineering strain for heterologous expression of cellulase gene EG IV and application
US20210284983A1 (en) Beta glucosidase with high glucose tolerance, high thermal stability and broad ph activity spectrum
Fiedurek et al. Seeds as natural matrices for immobilization of Aspergillus niger mycelium producing pectinases
CN106399337B (en) It is a kind of produce alkaline pectase recombinant bacterium and its application
CN113913410B (en) Construction and application of yak rumen anaerobic fungus xylanase gene engineering bacteria

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into 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: 20160713