CN106868026A - For producing the gene with temperature tolerance lipase - Google Patents
For producing the gene with temperature tolerance lipase Download PDFInfo
- Publication number
- CN106868026A CN106868026A CN201710177365.5A CN201710177365A CN106868026A CN 106868026 A CN106868026 A CN 106868026A CN 201710177365 A CN201710177365 A CN 201710177365A CN 106868026 A CN106868026 A CN 106868026A
- Authority
- CN
- China
- Prior art keywords
- lipase
- gene
- temperature tolerance
- ttg
- gtc
- 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
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
- C12N9/14—Hydrolases (3)
- C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
- C12N9/18—Carboxylic ester hydrolases (3.1.1)
- C12N9/20—Triglyceride splitting, e.g. by means of lipase
-
- A—HUMAN NECESSITIES
- A21—BAKING; EDIBLE DOUGHS
- A21D—TREATMENT, e.g. PRESERVATION, OF FLOUR OR DOUGH, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS; PRESERVATION THEREOF
- A21D2/00—Treatment of flour or dough by adding materials thereto before or during baking
- A21D2/08—Treatment of flour or dough by adding materials thereto before or during baking by adding organic substances
- A21D2/24—Organic nitrogen compounds
-
- A—HUMAN NECESSITIES
- A21—BAKING; EDIBLE DOUGHS
- A21D—TREATMENT, e.g. PRESERVATION, OF FLOUR OR DOUGH, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS; PRESERVATION THEREOF
- A21D8/00—Methods for preparing or baking dough
- A21D8/02—Methods for preparing dough; Treating dough prior to baking
- A21D8/04—Methods for preparing dough; Treating dough prior to baking treating dough with microorganisms or enzymes
- A21D8/042—Methods for preparing dough; Treating dough prior to baking treating dough with microorganisms or enzymes with enzymes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y301/00—Hydrolases acting on ester bonds (3.1)
- C12Y301/01—Carboxylic ester hydrolases (3.1.1)
- C12Y301/01003—Triacylglycerol lipase (3.1.1.3)
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Food Science & Technology (AREA)
- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- Enzymes And Modification Thereof (AREA)
Abstract
The present invention relates to biological technical field, specifically disclose a kind of for producing the gene with temperature tolerance lipase and the temperature tolerance lipase as expressed by the gene.Lipase temperature tolerance expressed by gene of the invention is good, with potentiality to be exploited, with extensive and actual application value, such as can be used for brightening for Flour product and baking goodses.
Description
Technical field
The invention belongs to biological technical field, and in particular to a kind of for producing the gene with temperature tolerance lipase.
Background technology
Lipase (Lipase, GEH) is under the jurisdiction of carboxylic ester hydrolase class, can be progressively by triglycerides
It is hydrolyzed into glycerine and aliphatic acid.Lipase is present in animal and plant and microorganism (such as mould, bacterium) tissue containing fat
In.Including phosphate, sterol enzyme and carboxy-lesterase.Aliphatic acid is widely used in food, medicine, leather, daily-use chemical industry etc.
Aspect.
However, in the application of lipase, its active temperature influence is larger, and when temperature is higher, the enzyme activity urgency of lipase
Play declines, and influences its commercialization to promote or apply.
The content of the invention
In order to solve the above problems, an object of the present invention there are provided a kind of for producing with temperature tolerance fat
The gene of enzyme；The second object of the present invention is to provide a kind of temperature tolerance lipase as expressed by the gene.
The present invention is achieved through the following technical solutions：
It is a kind of for producing the gene with temperature tolerance lipase, the sequence of the gene is as follows：
A kind of temperature tolerance lipase as expressed by said gene.
It is preferred that enzyme activity of the temperature tolerance lipase after being processed 50~90 minutes at 50~60 DEG C for 87.5%~
90.6%.
Using upper, the temperature tolerance lipase can be used to brighten Flour product, such as noodles, or brighten baking goodses, such as steamed bun
Deng.
Compared with prior art, the invention has the advantages that：Lipase heatproof expressed by gene of the invention
Property it is good, with potentiality to be exploited, with extensive and actual application value, such as can be used for brightening for Flour product and baking goodses.
Brief description of the drawings
Fig. 1 is the agarose electrophoresis detection of linearization plasmid APYLL-pPICZ α A of the present invention, wherein, line M, DL15000bp
marker；Line 2, linearisation APYLL-pPICZ α A.
Fig. 2 is the schematic diagram that BMMY- rhodamine Bs flat band method screens recon.
Specific embodiment
With reference to specific embodiment, the present invention is described in further detail, is managed to help those skilled in the art
The solution present invention.
The present invention is as follows for producing the sequence of the gene (being represented with APYLL-pPICZ α A) with temperature tolerance lipase：
First, the expression of gene
Host：Pichi strain X-33 and GS115.Certainly, other hosts that can express lipase gene can apply
To the present invention.
Gene chemical synthesis：The present invention is synthesized for producing the gene with temperature tolerance lipase by Shanghai JaRa company.
1.1 expression strain constructions
1.1.1 plasmid extraction
With reference to the Guangzhou small extraction reagent kit operation instructions of Dongsheng plasmid.
1.1.2 the linearisation and recovery of recombinant plasmid APYLL-pPICZ α A
The recombinant plasmid APYLL-pPICZ α A of gained are carried out into linearization process with Sac I, endonuclease reaction 1h at 37 DEG C,
Reaction system is as follows：
37 DEG C of water-baths are positioned over after endonuclease reaction solution is mixed, are taken out after 1h, carried out with gel-purified QIAquick Gel Extraction Kit
Reclaim (referring to Tiangeng glue reclaim kit operation instructions).
1.1.3 the preparation of Pichi strain competence and electricity conversion
Referring to pPICZ α A, B, and C user's operation manuals of Invitrogen.
1.1.4 result
Correct recombinant plasmid APYLL-pPICZ α A will be linearized with Sac I after digestion verification and sequence verification
Treatment, digestion products condense electrophoresis after reclaiming with 1% agarose to be identified, as a result as shown in Figure 1.
The screening of the transformant of 1.2 tool lipase actives
1.2.1 primary dcreening operation
Most fast monoclonal is grown on picking YPDS flat boards, and point is connected to YPD (for conservation) and BMMY- rhodamine Bs respectively
Flat board (for inducing) flat board, 30 DEG C are inverted incubated 3~5d；
100 μ L methyl alcohol are added on the lid of BMMY- rhodamine B flat boards every 24h carries out induced expression to recon,
In incubation, the relative size according to hydrolysis circle selects 10 most strong transformants of lipase enzymatic activity, and number consecutively is 1#,
2#, 3#~10#, plate screening result are as shown in Figure 2.
1.2.2 secondary screening
10 selected transformants are inoculated with (100mL specifications shaking flask), 30 DEG C in the fluid nutrient mediums of BMGY containing 10mL respectively
250rpm is cultivated to OD600=6.0；
3,000g centrifugation 5min collects thallines, with being transferred to 50mLBMMY liquid after 2mLBMMY fluid nutrient medium re-suspended cells
In body culture medium, make its OD600=1.0；
30 DEG C of 250rpm continue Fiber differentiation, are added after a methyl alcohol makes its final concentration of 0.5%, 48h every 24h and started
Enzymatic activity is surveyed in sampling, and sampling time point is 48h, 72h, 96h, 120h, and samples taken collects supernatant after 3,000g centrifugations 3min,
Lipase enzymatic activity is surveyed using titration, it is determined that active highest transformant and its inductive condition.
1.2.3 result
10 transformants that the lipase active of selection primary dcreening operation acquisition is higher are further screened, and observe its triangular flask water
Flat expression, using the lipase active in alkaline titration measuring fermented liquid supernatant, the enzyme activity of APYLL-pPICZ α A is surveyed
Surely the results are shown in Table 1.
The lipase enzymatic activity of each transformant in the horizontal triangular flask of the shaking table of table 1
| Induction time (h) | APYLL-pPICZ α A enzymatic activitys (U/mL) |
| 48 | 42 |
| 72 | 55 |
| 96 | 57 |
| 120 | 55 |
1.3 lipase activities are determined
This experiment uses alkali formula titration measuring lipase activity.
1.3.1 principle
Lipase under certain condition, can make triglyceride be hydrolyzed into aliphatic acid, diglyceride, monoglycerides and glycerine,
The aliphatic acid available standards aqueous slkali for being discharged carries out acid-base titration, is counted with PH or phenol peptide indicator solution Indicator Reaction terminal, according to
The alkali number of consumption, calculates its enzyme activity.Reaction equation is：
RCOOH+NaOH→RCOONa+H2O
1.3.2 operating procedure
1) two 50ml triangular flasks are taken, substrate solution 4.00ml olives is added respectively at each in blank tube (A) and sample cell (B)
Olive oil emulsion (olive oil：PVA=1:3, v/v) and 5.00ml phosphate buffers (100mM, pH 7.0), added in A pipes
95% ethanol 15.00ml, preheats 5min in 40 DEG C of water-baths, and enzyme liquid 1.00ml to be measured is then respectively added in A, B pipe, mixes immediately
Timing, the accurate response 10min in 40 DEG C of water-baths adds 95% ethanol 15.0ml terminating reactions immediately in B pipes, takes out；
2) respectively add phenol peptide indicator solution 2 to drip in blank and sample solution, titrated with 50mM NaOH standard liquids, until micro-
It is red and to keep 30s colour-fast be titration end-point, the volume of record consumption 50mM NaOH standard liquids.
1.3.3 enzyme activity is calculated
Enzyme activity is defined:1g solid enzyme powders (or 1mL liquid enzymes), under the conditions of 40 DEG C of temperature and pH 7.0,1min hydrolysis olives
Olive oil (olive oil) produces 1 μm of titratable aliphatic acid of ol, as 1 enzyme activity unit to be represented with U/g (U/ml).
Computational methods：The enzyme activity of lipase preparation is calculated as follows
X1=(V1-V2) * c*50*n1/0.05*1/10
In formula：
The enzyme activity of X1----- samples, U/g (U/ml)
The volume of consumption standard solution of sodium hydroxide during V1---- titration samples, unit is milliliter (ml)；
The volume of consumption standard solution of sodium hydroxide during V2---- titration blank, unit is milliliter (ml)
C---- Concentration of Sodium Hydroxide Solution Standard, unit is mole every liter (mol/L)；
50---0.05mol/L sodium hydroxide solutions 1.00ml is equivalent to 50 μm of ol of aliphatic acid；
The extension rate of n1---- samples；
0.05---- Concentration of Sodium Hydroxide Solution Standard conversion coefficients；
1/10---- reaction time 10min, in terms of 1min.
Acquired results are represented to integer.
1.3.4 temperature tolerance test
With above-mentioned Determination Methods for Lipase Activity test result.
Comparative example：LBK-B4000 (the special lipase of steamed bun, green micro- health)：46.5mg albumen/g powder (protein contents
4.65%)；Lipozyme TL 100L (are used for fats and oils processing, Novozymes)：24.2mg albumen/mL original enzyme liquid (protein contents
2.42%)；CaL-B (is used for organic synthesis, Novozymes)：9.24mg albumen/mL original enzyme liquids (protein content 0.924%).
By APYLL-pPICZ α A fermented supernatant fluids, the dilution (100 times of dilution) of LBK-B4000, Lipozyme TL
The dilution (dilution 100 times) of 100L and the original enzyme liquid of CaL-B be placed in 50 DEG C~60 DEG C at warm bath (60 DEG C are taken in the present embodiment),
Sample is gone to survey remnant enzyme activity respectively at 10,20,30,40,50,60,90 and 135min, the enzyme activity with Preliminary fermentation supernatant is
100%.Test result is as shown in table 2 below：
Table 2：Each lipase enzyme exists:Stability at 60 DEG C
As shown in Table 2：The temperature tolerance of APYLL-pPICZ α A is greatly improved, and the stability at 60 DEG C is more than 135 points
Clock, LBK-B4000 is 90 minutes in the stability at 60 DEG C, and Lipozyme TL 100L are 60 points in the stability at 60 DEG C
Clock, CaL-B is 60 minutes in the stability at 60 DEG C.In terms of temperature tolerance, our the lipase A PYLL-pPICZ α A in research
With potentiality to be exploited.
2nd, apply
By the way that Gene A PYLL-pPICZ α A of the invention are obtained into durothermic lipase on yeast after high efficient expression,
Two sample Fs NL-25000 and FSL-20000 are obtained by existing formulation optimization again, can be applied to respectively Flour product with
Baking goodses are brightened.Wherein, lipase optimization is belonged into prior art with suitable for Flour product or baking goodses, herein not
Repeat again.
2.1 lipase FNL-25000 noodles whitening effects are tested
2.1.1, experiment purpose
Whitening effects of the test lipase FNL-25000 in white salt noodles
2.1.2 primary raw material is tested
Flour：Muscle basis powder in Shandong
Lipase：FNL-25000
2.1.3 experimental formula and technique
Table 3, noodle formula
| Raw material | Consumption/g |
| Flour | 100 |
| Salt | 1 |
| Water | 33 |
Noodles technique：Add water and face；It is lax；Oodle maker pressure surface；Integer, is made noodles and dough sheet.
2.1.4 experimental result
Table 4, FNL-25000 test results
As can be known from the above table, with the increase of FNL-25000 concentration, noodles and dough sheet whiteness are incremented by successively.
2.1.5 experiment conclusion
Muscle basis powder adds lipase FNL-25000 as flour material with Shandong, and noodles whiteness substantially increases, and adds
Amount is bigger, and noodles whiteness is bigger.
2.2 lipase FSL-20000 steamed buns whitening effects are tested
2.2.1, experiment purpose
Test lipase FSL-20000 whitening effects in north steamed bread
2.2.2 primary raw material is tested
Flour：Muscle basis powder in Shandong；Muscle basis powder in Hebei
Lipase：FSL-20000
2.2.3 experimental formula and technique
Table 5, steamed bun formula
| Raw material | Consumption/g |
| Flour | 100 |
| Yeast | 0.8 |
| Water | 46 |
2.2.4 steamed bun technique
Add water to be rubbed with the hands with face → oodle maker pressure surface → craft and justify → proof → boiling
2.2.5 experimental result
Table 6, FSL-20000 test results
Muscle basis powder adds the increase of concentration with FSL-20000 in muscle basis powder and Hebei in Shandong, and steamed bun epidermis is white
Degree increases successively.
2.2.6 conclusion
With in Shandong muscle basis powder and Hebei in muscle basis powder as flour material, with the addition of lipase FSL-20000, steamed bun
Epidermis whiteness substantially increases, and with the increase of addition, steamed bun epidermis whiteness is in rising trend.
Applicant：Dongguan Fanyatai Biological Sci-Tech Co., Ltd.
Denomination of invention：For producing the gene with temperature tolerance lipase
Sequence signature：Gene complete sequence for producing the gene with temperature tolerance lipase
Sequence description：
GAA TTC GTC TAC ACC TCT ACC GAG ACC TCT CAC ATC GAC CAG GAG TCT TAC AAC
TTC TTC GAG AAG TAC GCC AGA TTG GCC AAT ATC GGT TAC TGC GTC GGA CCT GGA ACC
AAG ATT TTC AAG CCA TTC AAC TGC GGT TTG CAA TGC GCA CAC TTC CCA AAC GTC GAG
TTG ATC GAG GAG TTC CAT GAC CCT AGA TTG ATC TTT GAC GTC TCT GGT TAC TTG GCC
GTC GAC CAC GCA TCT AAG CAA ATC TAC TTG GTC ATC AGA GGA ACA CAC TCT TTG GAG
GAT GTC ATC ACC GAC ATT AGA ATC ATG CAG GCA CCA TTG ACC AAC TTC GAC TTG GCA
GCC AAC ATC TCT TCT ACA GCA ACC TGC GAC GAC TGT TTG GTC CAC AAC GGT TTC ATC
CAG TCT TAC AAC AAC ACC TAC AAT CAA ATC GGT CCA AAG TTG GAC TCT GTC ATC GAG
CAG TAC CCT GAT TAC CAA ATC GCT GTT ACT GGT CAC TCT TTG GGT GGT GCT GCC GCC
TTG TTG TTC GGT ATC AAC TTG AAG GTC AAC GGT CAC GAC CCT TTG GTC GTT ACC TTG
GGA CAA CCA ATT GTC GGA AAC GCT GGT TTC GCC AAC TGG GTC GAC AAG TTG TTC TTC
GGT CAA GAA AAC CCT GAC GTT TCT AAG GTC TCT AAG GAC AGA AAG TTG TAC AGA ATC
ACC CAC AGA GGA GAT ATC GTC CCA CAG GTC CCT TTT TGG GAC GGA TAC CAG CAT TGC
TCT GGT GAG GTT TTC ATC GAC TGG CCT TTG ATT CAC CCT CCT TTG TCT AAC GTC GTC
ATG TGC CAG GGT CAG TCT AAC AAG CAA TGT TCT GCC GGT AAC ACA TTG TTG CAG CAG
GTT AAT GTC ATC GGT AAC CAC TTG CAA TAC TTC GTC ACT GAG GGT GTC TGC GGA ATT
TAA GCG GCC GC。
Claims (4)
1. it is a kind of for producing the gene with temperature tolerance lipase, it is characterised in that the sequence of the gene is as follows：
2. the temperature tolerance lipase expressed by a kind of gene as described in claim 1.
3. temperature tolerance lipase according to claim 2, it is characterised in that the temperature tolerance lipase in 50~60 DEG C
It is lower treatment 50~90 minutes after enzyme activity be 87.5%~90.6%.
4. temperature tolerance lipase according to claim 2, it is characterised in that the temperature tolerance lipase is used for whitening face system
Product and baking goodses.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710177365.5A CN106868026A (en) | 2017-03-22 | 2017-03-22 | For producing the gene with temperature tolerance lipase |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710177365.5A CN106868026A (en) | 2017-03-22 | 2017-03-22 | For producing the gene with temperature tolerance lipase |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106868026A true CN106868026A (en) | 2017-06-20 |
Family
ID=59171949
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710177365.5A Pending CN106868026A (en) | 2017-03-22 | 2017-03-22 | For producing the gene with temperature tolerance lipase |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106868026A (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20060024789A1 (en) * | 2004-08-02 | 2006-02-02 | Rahman Raja N Z | Lipase from Geobacillus sp. strain T1 |
| CN102169532A (en) * | 2010-12-10 | 2011-08-31 | 江南大学 | Correlation computing of amino acid and dipeptide thereof with optimal temperature of lipase |
| CN104152471A (en) * | 2014-08-22 | 2014-11-19 | 武汉轻工大学 | Lipase gene COLIP and lipase encoded by same |
| CN105264070A (en) * | 2014-02-26 | 2016-01-20 | 江南大学 | New bifunctional lipase mutant and uses thereof in flour product processing |
| CN105950585A (en) * | 2016-04-29 | 2016-09-21 | 华南农业大学 | Thermally stable lipase as well as preparation method and applications thereof |
-
2017
- 2017-03-22 CN CN201710177365.5A patent/CN106868026A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20060024789A1 (en) * | 2004-08-02 | 2006-02-02 | Rahman Raja N Z | Lipase from Geobacillus sp. strain T1 |
| CN102169532A (en) * | 2010-12-10 | 2011-08-31 | 江南大学 | Correlation computing of amino acid and dipeptide thereof with optimal temperature of lipase |
| CN105264070A (en) * | 2014-02-26 | 2016-01-20 | 江南大学 | New bifunctional lipase mutant and uses thereof in flour product processing |
| CN104152471A (en) * | 2014-08-22 | 2014-11-19 | 武汉轻工大学 | Lipase gene COLIP and lipase encoded by same |
| CN105950585A (en) * | 2016-04-29 | 2016-09-21 | 华南农业大学 | Thermally stable lipase as well as preparation method and applications thereof |
Non-Patent Citations (4)
| Title |
|---|
| GEORGES PIGNÈDE ET AL.: "Characterization of an Extracellular Lipase Encoded by LIP2 in Yarrowia lipolytica", 《JOURNAL OF BACTERIOLOGY》 * |
| ZHOU,W. ET AL.: "ACCESSION NO：AIB08846，lipase 2, partial [synthetic construct]", 《GENBANK》 * |
| ZHOU,W.: "ACCESSION NO：AIB08847，lipase 2, partial [synthetic construct]", 《GENBANK》 * |
| 季祥 等: "《生物质能源及废物利用新技术》", 31 December 2012 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Hernández et al. | Saccharification of carbohydrates in microalgal biomass by physical, chemical and enzymatic pre-treatments as a previous step for bioethanol production | |
| CN104185680B (en) | Method for producing ethanol and fermented solid product simultaneously | |
| CN103829036A (en) | Preparation method of microecological fermented feed by utilizing byproducts of corn deep-processing as raw materials | |
| Yang et al. | Fermentation of rice hull by Aspergillus japonicus under ultrasonic pretreatment | |
| CN109486867A (en) | Compound cellulose enzyme system and its application in starch fuel ethanol production | |
| Gupta et al. | Optimization of xylanase production from Melanocarpus albomyces using wheat straw extract and its scale up in stirred tank bioreactor | |
| Ben Rejeb et al. | Bread surplus: a cumulative waste or a staple material for high-value products? | |
| CN104789492B (en) | Bacillus megaterium bacterial strain and its application | |
| CN104789491B (en) | Lichem bacillus strain and its application | |
| CN104498575A (en) | Beer yeast polypeptide with anti-oxidation effect and preparation method thereof | |
| CN106868026A (en) | For producing the gene with temperature tolerance lipase | |
| CN101735331B (en) | Production process for extracting lily polysaccharides through fermentation method and product thereof | |
| CN102994471B (en) | Lipase mutant with increased optimum temperature and application of lipase mutant with increased optimum temperature | |
| CN108771134A (en) | A kind of method of soluble dietary fiber in enzyme assisted extraction bean dregs | |
| CN105483169A (en) | Method for producing aliphatic acid through utilization of gutter cooking oil in enzymic method | |
| CN107460223A (en) | A kind of degreasing silkworm chrysalis hydrolysate for microculture and its preparation method and application | |
| CN105400704B (en) | A method of improving the dregs of a decoction anaerobic fermentation efficiency of antibiotic containing sporinite | |
| CN106967737A (en) | For producing the gene with high activity lipase | |
| CN104046542B (en) | The method of edible ethanol is produced with inferior red date thick mash thermophilic fermentation | |
| CN106434771B (en) | A method of utilizing aqueous enzymatic method hydrolyzate, residue fermenting and producing alcohol | |
| CN102653721A (en) | Method for preparing monascus mycelia with function of reducing blood fat by liquid state fermentation of grains | |
| CN109182421A (en) | A kind of method that solid state fermentation brewex's grains prepare feruloylated oligosaccharides and dietary fiber | |
| CN105018364B (en) | One plant of saccharomyces cerevisiae engineered yeast for expressing pectinesterase and its application | |
| CN104846047A (en) | Method for preparing zein antihypertensive peptide based on ultrasonic reverberant field | |
| CN104762166A (en) | Fed-batch fermentation method for preparing high-degree yellow wine |
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 |
Application publication date: 20170620 |
|
| RJ01 | Rejection of invention patent application after publication |