CN104651391A - Recombinant expression and application of lactobacillus casei phospholipase A2 gene - Google Patents
Recombinant expression and application of lactobacillus casei phospholipase A2 gene Download PDFInfo
- Publication number
- CN104651391A CN104651391A CN201510078380.5A CN201510078380A CN104651391A CN 104651391 A CN104651391 A CN 104651391A CN 201510078380 A CN201510078380 A CN 201510078380A CN 104651391 A CN104651391 A CN 104651391A
- Authority
- CN
- China
- Prior art keywords
- phospholipase
- gene
- recombinant
- expression
- enzyme
- 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
Landscapes
- Enzymes And Modification Thereof (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to a recombinant expression method of a lactobacillus casei phospholipase A2 gene. The recombinant expression method of the lactobacillus casei phospholipase A2 gene sequentially comprises the following steps: predicting a signal peptide of the gene, constructing a recombinant carrier, converting, screening, detecting an expression product, producing enzyme by recombinant bacteria, and fermenting; concretely firstly predicting the signal peptide of the gene according to the sequence of the lactobacillus casei phospholipase A2 gene, introducing a restriction enzyme cutting site at the two ends of a mature peptide by utilizing a specific primer, subcloning the mature peptide into a carrier pKLAC1 to obtain an expression carrier pKLAC1-pla2; then screening a recombinant lactic acid kluwe yeast strain to realize expression of recombinant phospholipase A2; then collecting supernate obtained through fermentation, and primarily detecting enzyme activity by adopting an egg yolk plate; and finally carrying out shaking fermentation cultivation on yeast. The recombinant expression method of the lactobacillus casei phospholipase A2 gene has the advantages that an eukaryotic expression vector is constructed according to the cloned lactobacillus casei phospholipase A2 gene, the lactobacillus casei phospholipase A2 gene is expressed by utilizing a yeast expression system, and influence of different fermentation conditions on growth of recombinant strains and enzyme production is studied.
Description
Technical field
The present invention relates to technical field of molecular biology, especially relate to the phospholipase A in a kind of lactobacterium casei source
2recombinant expressed and the application of gene.
Background technology
Phospholipase A
2, English phospholipase A by name
2, be abbreviated as PLA
2, it can generate lysolecithin and free fatty acids by catalyze phospholipid glycerol molecule two acyl hydrolases, and lysolecithin has important application in food, makeup and field of medicaments, and Japan it can be used as natural additive for foodstuff.Therefore, it is a kind of important enzyme in industrial production.
Phospholipase A
2extensively be present in various organism.It is by extracting directly in biological tissue the earliest, but there is the problems such as extraction yield is low, complicated operation.Some mammal phospholipase As
2gene is carried out recombinant expressed by trial in the Host Strains such as intestinal bacteria, aspergillus, but exist form inclusion body, expression amount is low does not reach the problems such as industrial applications, meanwhile, utilizes these expressive hosts to express the mammalian phospholipase A of some complexity
2can be there are some restrictions in gene, as completed the post-treatment of corresponding recombinant protein, the genetic background of these expressive host bacterium can limit the related application etc. of recombinase, in addition, studies have found that recombinant expressed phospholipase A
2can to intestinal bacteria, the expressive host generation toxic actions such as yeast, thus limit recombinant expressed and application accordingly.And by microbe-derived phospholipase A
2gene carries out recombinant expressed in Kluyveromyces lactis, effectively can improve the problems referred to above.Now both at home and abroad not about by microbe-derived phospholipase A
2gene carries out recombinant expressed report in Kluyveromyces lactis.
Summary of the invention
For the problems referred to above that prior art exists, the applicant provides a kind of lactobacterium casei phospholipase A
2recombinant expressed and the application of gene.The present invention is according to the lactobacterium casei phospholipase A be cloned into
2gene, builds carrier for expression of eukaryon, utilizes yeast expression system to have expressed lactobacterium casei phospholipase A
2gene, and have studied different fermentations condition to restructuring strain growth and the impact of product enzyme.
Technical scheme of the present invention is as follows:
Lactobacterium casei phospholipase A
2the recombinant expression method of gene, comprise gene signal peptide prediction, the Gou Jian ﹑ conversion of recombinant vectors and the detection of Shai Xuan ﹑ expression product successively, recombinant bacterium produces enzymic fermentation;
The structure of described expression vector is: the lactobacterium casei phospholipase A announced according to NCBI
2gene order, signalP carries out gene signal peptide prediction, utilizes Auele Specific Primer to introduce restriction enzyme site at mature peptide two ends, is subcloned in carrier pKLAC1, obtains expression vector pKLAC1-pla2.;
Described conversion and screening are: transform Kluyveromyces lactis with the expression vector built, and screening recombination lactic acid kluyveromyces yeast strains realizes restructuring phospholipase A
2expression;
The detection of described expression product is: collect recombination lactic acid kluyveromyces fermented supernatant fluid, egg plate Preliminary detection enzyme is lived;
Described recombinant bacterium produces enzymic fermentation fermentation: carry out shake flask fermentation cultivation to restructuring Kluyveromyces lactis, fermentation condition is: glucose is 1%-3%, yeast powder is 1%-2%, peptone is 1%-3%, KH
2pO
4for 0.1%-0.3%, inoculum size are 1%-4% and liquid amount 70-100ml.The fermentation condition optimized is: glucose is 3%, yeast powder is 2%, peptone is 3%, KH
2pO
4be 0.3%, inoculum size is 2% and liquid amount 90ml.
Lactobacterium casei phospholipase A
2recombinant expressed in Kluyveromyces lactis of gene is secreting, expressing.
Described Auele Specific Primer is:
With lactobacterium casei genome for template, pla2-F/pla2-R is primer, primer two ends introduce Xho I and Bgl II restriction enzyme site respectively, and introduce Kex restriction enzyme site gene order at upstream region of gene simultaneously, amplify the lactobacterium casei phospholipase A not comprising signal peptide
2gene, i.e. described primer;
Described primer sequence is:
pla2-F：5’-CCGCTCGAGAAAAGAACGACTAAGACTGAGTTTAATG-3’；
pla2-R：5’-GGAAGATCTTCAACCACCAAACAACTTATATG-3’。
This restructuring phospholipase A
2be applied to food, makeup and medicine and other fields.
The technique effect that the present invention is useful is:
The present invention utilizes the lactobacterium casei phospholipase A_2 gene be cloned into build carrier for expression of eukaryon, to provide the recombinant expressed of Phospholipase A2 and application.Recombinant phospholipase A2 can be applicable to food, makeup and medicine and other fields.The present invention is by DNA recombinant technology, lactobacterium casei phospholipase A_2 gene is increased out, build recombinant expression vector, achieve recombinant expressed in Kluyveromyces lactis of lactobacterium casei phospholipase A_2 gene, and have studied different fermentations condition on restructuring strain growth and produce enzyme impact, for the suitability for industrialized production of Phospholipase A2 and application are laid a good foundation.
The present invention have expressed lactobacterium casei source Phospholipase A2 in Kluyveromyces lactis, and egg plate Preliminary detection enzyme is lived, and determination of acid-basetitration enzyme size alive, SDS-PAGE shows recombinant protein band, and instruction card reaches merit.In addition, optimized by shake flask fermentation, recombinant phospholipase A2 enzyme is lived and is reached and can reach 6.56U/ml, for the suitability for industrialized production of Phospholipase A2 is laid a good foundation.
Expression product of the present invention can be applicable to food, makeup and medicine and other fields, is with a wide range of applications.
Accompanying drawing explanation
Fig. 1: be lactobacterium casei Phospholipase A2 signal amino acid sequence peptide prognostic chart;
Fig. 2: be recombinant expression vector digestion verification figure;
Fig. 3: be that egg plate method detects recombinant bacterium fermentation supernatant Phospholipase A2 enzymic activity figure;
Fig. 4: the SDS-PAGE protein electrophoresis figure being recombinant bacterium fermentation supernatant expression product;
Fig. 5: be recombinant bacterium GG799/pKLAC1-pla2 Fermentative growth and product enzyme curve;
Fig. 6: be that calcium ion concn is to recombinase Phospholipase A2 enzyme effect diagram alive;
Fig. 7: be that carbon source is to GG799/pKLAC1-pla2 growth and the effect diagram producing enzyme;
Fig. 8: be that nitrogenous source is to GG799/pKLAC1-pla2 growth and the effect diagram producing enzyme;
Fig. 9: be that inorganic salt are to GG799/pKLAC1-pla2 growth and the effect diagram producing enzyme;
Figure 10: be that inoculum size is to GG799/pKLAC1-pla2 growth and the effect diagram producing enzyme;
Figure 11: be that initial pH is to GG799/pKLAC1-pla2 growth and the effect diagram producing enzyme;
Figure 12: be that liquid amount is to GG799/pKLAC1-pla2 growth and the effect diagram producing enzyme.
Embodiment
Below in conjunction with accompanying drawing, the present invention is specifically described.Wherein to recombinate phospholipase A
2purchased from Southern Yangtze University's Chinese Universities ' industrial microorganism resource and information center, Kluyveromyces lactis is purchased from Niu Yinglun Bioisystech Co., Ltd.
Kluyveromyces lactis GG799 expression system is utilized to express restructuring phospholipase A
2.
(1) lactobacterium casei phospholipase A
2the prediction of gene signal peptide:
NCBI checks in lactobacterium casei phospholipase A
2the aminoacid sequence of gene order and corresponding encoded, SignalP 4.1server (http://www.cbs.dtu.dk/services/SignalP/) carries out signal peptide prediction to aminoacid sequence, predicts the outcome as shown in Figure 1.
(2) structure of recombinant expression vector pKLAC1-pla2:
Predicting the outcome according to signal peptide, with lactobacterium casei genome for template, pla2-F/pla2-R is primer, primer two ends introduce Xho I and Bgl II restriction enzyme site site respectively, and introduce Kex restriction enzyme site gene order at upstream region of gene simultaneously, amplify the lactobacterium casei phospholipase A not comprising signal peptide
2gene.By carrier pKLAC1 and goal gene Xho I and Bgl II respectively enzyme cut, glue reclaim, in 16 DEG C of connections of spending the night, Transformed E .coli JM109.Then, extract JM109/pKLAC1-pla2 bacterial strain plasmid, Xho I and Bgl II digestion verification, the result as shown in Figure 2, filters out JM109/pKLAC1-pla2 positive strain.
(3) linearization plasmid pKLAC1-pla2 electricity transforms the screening of Kluyveromyces lactis GG799 and transformant:
Extract JM109/pKLAC1-pla2 positive strain plasmid, after Sac II linearizing, electricity transforms Kluyveromyces lactis GG799, and coating YCB is dull and stereotyped, and ethanamide screens, 3-4 days picking list bacterium colonies.Extract recombinant bacterium chromogene group by glass bead method, utilize integration primer (see table 1) to carry out PCR checking, filter out the multiple copied recombinant bacterium of Successful integration goal gene.
Table 1 primer
(4) expression of recombinase in Kluyveromyces lactis Kluyveromyces lactis GG799:
The positive recombinant bacterium screened is seeded in 50mL YPD substratum, cultivates 24h, as seed liquor, be forwarded in 50ml YPD substratum with 2% inoculum size, cultivate in 30 DEG C ﹑ 200r/min condition bottom fermentations, every 24h sampling, egg plate Preliminary detection phospholipase A
2enzyme is lived, and as shown in Figure 3, wherein numbering 0 is contrast; And by SDS-PAGE, fermentation supernatant expressing protein is analyzed, as shown in Figure 4; Determination of acid-basetitration phospholipase A
2enzyme is lived, and to determine its suitableeest enzymatic production time, as shown in Figure 5, recombinant bacterium is lived at 72h enzyme and reached maximum, and enzyme is lived as 1.87U/ml; Different calcium ionic concn lives impact as shown in Figure 6 to Phospholipase A2 enzyme.
(5) optimization of recombinant bacterium conditions of flask fermentation:
Investigate different carbon source to restructuring bacteria growing and the impact of producing enzyme.With 2% (m/v) peptone and 1% yeast powder for nitrogenous source, shake flask fermentation is carried out to recombinant bacterium with breast sugar ﹑ semi-lactosi, sugarcane sugar ﹑ glucose, sweet oily ﹑ maltose and the dextrin of 2% for carbon source respectively, 72h surveys thalli growth amount and enzyme is lived, as shown in Figure 7.Result shows, is conducive to phospholipase A with lactose, semi-lactosi and glucose for during carbon source
2expression, enzyme live all compared with high during other carbon source.When taking dextrin as carbon source, biomass and enzyme viable bacteria lower, illustrate that recombinant bacterium can not utilize dextrin preferably, and with sucrose, glycerine and maltose for carbon source is conducive to the growth of thalline, but be unfavorable for the expression of recombinase, and when carbon source is glucose, enzyme work is the highest in selected carbon source, therefore, glucose is chosen as carbon source.
Investigate different nitrogen sources to restructuring bacteria growing and the impact of producing enzyme.With 2% glucose for carbon source, respectively with single nitrogenous source and peptones such as 1% fish meal protein Dong ﹑ yeast Fen ﹑ Yi Dan Bai Dong ﹑ beef extract, SODIUMNITRATE and ammonium chlorides: yeast extract paste (2:1) compound nitrogen source is that nitrogenous source carries out shake flask fermentation, 72h surveys thalli growth amount and enzyme is lived, as shown in Figure 8.As seen from the figure, organic nitrogen source is to the growth of thalline and produce enzyme all advantageously, and recombinant bacterium to inorganic nitrogen-sourced utilize poor, the enzyme work of compound nitrogen source is apparently higher than single nitrogenous source, peptone and yeast powder are that enzyme is lived the highest as nitrogenous source simultaneously, therefore, peptone and yeast powder compound nitrogen source is chosen as fermentation nitrogen source.
Investigate inorganic salt to restructuring bacteria growing and the impact of producing enzyme.Take glucose as carbon source, peptone and yeast powder are nitrogenous source, respectively with KH
2pO
4, MgCl
2, MnCl
2, ZnCl
2, FeCl
2, NaCl, NiCl
2and CaCl
2for unique inorganic salt carry out shake flask fermentation to recombinant bacterium, 72h surveys thalli growth amount and enzyme is lived, as shown in Figure 9.Discovery is compared, ZnCl with control group
2, FeCl
2restraining effect is had to product enzyme, but little on thalli growth impact, and KH
2pO
4be conducive to recombinant bacterium and produce enzyme, therefore, select KH
2pO
4as the inorganic salt of fermention medium.
Investigate inoculum size to restructuring bacteria growing and the impact of producing enzyme.Be that (inoculum size of v/v) ﹑ 2% ﹑ 3% ﹑ 4% ﹑ 5% is received in 50mL fermention medium, and after 72h, sampling surveys thalli growth amount and enzyme is lived, as shown in Figure 10 with 1% respectively for the seed liquor of 22h by kind of age.As seen from the figure, under this condition, inoculum size is little on the impact of strain growth amount, but inoculum size 3% and above time, be unfavorable for that recombinant bacterium produces enzyme, when inoculum size is 2%, enzyme is lived the highest, and therefore, choosing inoculum size is 2%.
Investigate the initial pH of substratum to restructuring bacteria growing and the impact of producing enzyme.Regulate the initial pH value of fermention medium to be respectively 4.0,5.0,6.0,7.0 and 8.0, shake flask fermentation, 72h surveys thalli growth amount and enzyme is lived, as shown in figure 11.Result shows, the tolerance range of recombinant bacterium to pH is wider, within the scope of selected pH, strain growth is substantially unaffected, and when pH value is 8, enzyme is lived lower, be unfavorable under this condition is described that recombinant bacterium produces enzyme, and pH value is when being 7.0, be conducive to producing enzyme, under this condition, enzyme is lived the highest, therefore, fermention medium initial pH value is regulated to be 7.0.
Investigate liquid amount to restructuring bacteria growing and the impact of producing enzyme.In 250ml triangular flask, be respectively charged into 40ml, 50ml, 60ml, 70ml and 80ml fermention medium, shake flask fermentation, 72h surveys thalli growth amount and enzyme is lived, as shown in figure 12.Result shows, liquid amount all has impact to strain growth and product enzyme, and larger on the impact of product enzyme.When liquid amount is 40ml, be unfavorable for that recombinant bacterium produces enzyme, along with the increase of liquid amount, enzyme is lived and is improved gradually, and when liquid amount is 70ml, enzyme is lived and reached the highest, and when liquid amount is increased to 80ml, enzyme is lived and reduced, and therefore, choosing shaking flask liquid amount is 70ml.
According to the experiment of single factor result of orthogonal design principle and fermention medium and fermentation condition, adopt L
18(3
7) carrying out the orthogonal test of six factor three levels, level of factor is in table 2, and experimental result and range analysis are in table 3.
Table 2 orthogonal test factor and level design
Table 3 orthogonal experiments and range analysis
From table 2,3, each factor on the size that affects that recombinant bacterium fermenting enzyme is lived is: KH
2p0
4> glucose > liquid amount > yeast powder > inoculum size > peptone, three different levelss of more each factor, obtaining best of breed is A
2b
3c
3d
3e
2f
2.Namely each factor addition glucose is 3%, yeast powder is 2%, peptone is 3%, KH
2pO
4be 0.3%, inoculum size is 2% and liquid amount 90ml.Carry out proof test under this condition, recombinase enzyme is lived and is reached 6.56U/ml.
Expression product of the present invention can be applicable to food, makeup and medicine and other fields, is with a wide range of applications.
Claims (5)
1. lactobacterium casei phospholipase A
2the recombinant expression method of gene, is characterized in that: comprise gene signal peptide prediction, the Gou Jian ﹑ conversion of recombinant vectors and the detection of Shai Xuan ﹑ expression product successively, recombinant bacterium produces enzymic fermentation;
The structure of described expression vector is: the lactobacterium casei phospholipase A announced according to NCBI
2gene order, signalP carries out gene signal peptide prediction, utilizes Auele Specific Primer to introduce restriction enzyme site at mature peptide two ends, is subcloned in carrier pKLAC1, obtains expression vector pKLAC1-pla2;
Described conversion and screening are: transform Kluyveromyces lactis with the expression vector built, and screening recombination lactic acid kluyveromyces yeast strains realizes restructuring phospholipase A
2expression;
The detection of described expression product is: collect recombination lactic acid kluyveromyces fermented supernatant fluid, egg plate Preliminary detection enzyme is lived;
Described recombinant bacterium produces enzymic fermentation fermentation: carry out shake flask fermentation cultivation to restructuring Kluyveromyces lactis, fermentation condition is: glucose is 1%-3%, yeast powder is 1%-2%, peptone is 1%-3%, KH
2pO
4for 0.1%-0.3%, inoculum size are 1%-4% and liquid amount 70-100ml.
2. method according to claim 1, is characterized in that: lactobacterium casei phospholipase A
2recombinant expressed in Kluyveromyces lactis of gene is secreting, expressing.
3. method according to claim 1, is characterized in that: described Auele Specific Primer is:
With lactobacterium casei genome for template, pla2-F/pla2-R is primer, and primer two ends introduce Xho I and Bgl II restriction enzyme site respectively, and introduce Kex restriction enzyme site gene order at upstream region of gene simultaneously, amplify the lactobacterium casei phospholipase A not comprising signal peptide
2gene, i.e. described primer;
Described primer sequence is:
pla2-F：5’-CCGCTCGAGAAAAGAACGACTAAGACTGAGTTTAATG-3’；
pla2-R：5’-GGAAGATCTTCAACCACCAAACAACTTATATG-3’。
4. lactobacterium casei phospholipase A according to claim 1
2the recombinant expression method of gene, is characterized in that: the fermentation condition that described recombinant bacterium produces enzymic fermentation is: glucose is 3%, yeast powder is 2%, peptone is 3%, KH
2pO
4be 0.3%, inoculum size is 2% and liquid amount 90ml.
5. lactobacterium casei phospholipase A according to claim 1
2the recombinant expression method of gene, is characterized in that: restructuring phospholipase A
2be applied to food, makeup and medicine and other fields.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510078380.5A CN104651391A (en) | 2015-02-13 | 2015-02-13 | Recombinant expression and application of lactobacillus casei phospholipase A2 gene |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510078380.5A CN104651391A (en) | 2015-02-13 | 2015-02-13 | Recombinant expression and application of lactobacillus casei phospholipase A2 gene |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104651391A true CN104651391A (en) | 2015-05-27 |
Family
ID=53243034
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510078380.5A Pending CN104651391A (en) | 2015-02-13 | 2015-02-13 | Recombinant expression and application of lactobacillus casei phospholipase A2 gene |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104651391A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109722406A (en) * | 2018-11-21 | 2019-05-07 | 东北农业大学 | The thymidylate synthase gene defection type recombinant lactobacillus casei and its construction method of expression Bovine lactoferricin and application |
| WO2019196791A1 (en) * | 2018-04-09 | 2019-10-17 | 中国科学院青岛生物能源与过程研究所 | Recombinant yeast strain for producing nervonic acids and application thereof |
| EP3740079B1 (en) * | 2018-01-15 | 2024-02-21 | Chr. Hansen A/S | Fermented milk product and preparation thereof using phospholipase |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1156955A (en) * | 1995-05-15 | 1997-08-13 | 吉斯特·布罗卡迪斯股份有限公司 | Application of phospholipases in animal feed |
| CN102787087A (en) * | 2012-08-07 | 2012-11-21 | 江南大学 | Lactobacillus casei and application of lactobacillus casei to feed |
-
2015
- 2015-02-13 CN CN201510078380.5A patent/CN104651391A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1156955A (en) * | 1995-05-15 | 1997-08-13 | 吉斯特·布罗卡迪斯股份有限公司 | Application of phospholipases in animal feed |
| CN102787087A (en) * | 2012-08-07 | 2012-11-21 | 江南大学 | Lactobacillus casei and application of lactobacillus casei to feed |
Non-Patent Citations (5)
| Title |
|---|
| HUI WANG ET AL.: "Secretory expression of a phospholipase A2 from Lactobacillus casei DSM20011 in Kluyveromyces lactis", 《JOURNAL OF BIOSCIENCE AND BIOENGINEERING》 * |
| 刘丹丹: "Lactobacillus casei磷脂酶A2基因的克隆及重组表达", 《中国优秀硕士学位论文全文数据库基础科学辑》 * |
| 刘丹丹等: "Lactobacillus casei 磷脂酶A2基因在大肠杆菌中的重组表达", 《生物加工过程》 * |
| 延晋雷: "磷脂酶A_1产生菌的筛选_重组大肠杆菌的构建及自诱导发酵", 《中国优秀硕士学位论文全文数据库基础科学辑》 * |
| 王辉等: "重组乳酸克鲁维酵母产磷脂酶A2发酵条件的优化", 《食品科学》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP3740079B1 (en) * | 2018-01-15 | 2024-02-21 | Chr. Hansen A/S | Fermented milk product and preparation thereof using phospholipase |
| WO2019196791A1 (en) * | 2018-04-09 | 2019-10-17 | 中国科学院青岛生物能源与过程研究所 | Recombinant yeast strain for producing nervonic acids and application thereof |
| CN109722406A (en) * | 2018-11-21 | 2019-05-07 | 东北农业大学 | The thymidylate synthase gene defection type recombinant lactobacillus casei and its construction method of expression Bovine lactoferricin and application |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Wen et al. | Enhanced solvent production by metabolic engineering of a twin-clostridial consortium | |
| ES2428825T3 (en) | Cellulase production process based on the regulation of the oscillation of dissolved oxygen pressure in the culture medium | |
| CN103468624B (en) | Genetic engineering bacteria used for high efficient production of mycose | |
| CN104450561B (en) | One plant of application produced chitinase bacterial strain and its chitinase is produced using crab shell fermentation | |
| Sun et al. | Direct cloning, expression of a thermostable xylanase gene from the metagenomic DNA of cow dung compost and enzymatic production of xylooligosaccharides from corncob | |
| Tian et al. | Recent advances in lactic acid production by lactic acid bacteria | |
| CN113278644B (en) | Construction and fermentation method of high-yield artificial bacterial strain of fengycin | |
| Rodriguez-Gomez et al. | Examining the potential of plasma-assisted pretreated wheat straw for enzyme production by Trichoderma reesei | |
| CN109182423A (en) | Promote the method for Escherichia coli fermentation production poly sialic acid | |
| CN103571772A (en) | Novel strain for producing butanol and method for producing butanol | |
| Hu et al. | Efficient production of d-1, 2, 4-butanetriol from d-xylose by engineered Escherichia coli whole-cell biocatalysts | |
| CN104651391A (en) | Recombinant expression and application of lactobacillus casei phospholipase A2 gene | |
| CN104046586B (en) | One strain gene engineering bacterium and the application in producing (2R, 3R)-2,3-butanediol thereof | |
| Liu et al. | Metal-organic frameworks coupling simultaneous saccharication and fermentation for enhanced butyric acid production from rice straw under visible light by Clostridium tyrobutyricum CtΔack:: cat1 | |
| Zhang et al. | Consolidated bioprocessing for bioethanol production by metabolically engineered cellulolytic fungus Myceliophthora thermophila | |
| CN105062938A (en) | Engineering bacterium capable of producing D-lactate by aid of pentose and hexose synchronously by means of fermentation, and fabrication and application of engineering bacterium | |
| CN109251938A (en) | A kind of Pediococcus acidilactici construction method of common fermentation glucose and xylose production Pfansteihl | |
| JP2023534879A (en) | N-Acetylglucosamine-producing strain and method for construction and use thereof | |
| CN104152483A (en) | Application of argJ gene in fermentation production of L-citrulline | |
| CN107881123B (en) | Genetically engineered bacterium for producing pyruvic acid by using methanol and construction method and application thereof | |
| WO2021129396A1 (en) | Recombinant filamentous fungus for producing ethanol, and construction and application thereof | |
| CN103562216A (en) | Enhanced fermentation of cellodextrins and beta-D-glucose | |
| Ge et al. | Improvement of l-lactic acid production from orange peels in mixed culture system | |
| Yang et al. | Construction of a novel а-agglutinin expression system on the surface of wild-type Saccharomyces cerevisiae Y5 and genetic immobilization of β-glucosidase1 | |
| Silva et al. | High cell density co-culture for production of recombinant hydrolases |
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: 20150527 |