58 results about "Vitreoscilla hemoglobin" patented technology

The present invention discloses a high-yield gamma-polyglutamic acid production gene engineering bacterial and a fermentation production method thereof. The gene engineering bacterial is named Bacillus subtilis FRD518, wherein the preservation number is CGMCC NO.6772, and the Vitreoscilla hemoglobin gene (vgb) is recombined and integrated on the chromosome of the gene engineering bacterial, such that the Vitreoscilla hemoglobin VHb can be successfully and highly expressed so as to significantly improve the oxygen utilization rate of the recombinant Bacillus subtilis under a low dissolved oxygen condition. According to the present invention, during a fermentation process, a carbon source, sodium glutamate, a yeast extract and other components are added in a flow manner, such that the gene engineering bacterial can efficiently produce the gamma-polyglutamic acid in a high yield manner, wherein the yield achieves more than 65 g/L, and is increased by 147% compared with the yield of the single batch culture of the original wild strain; and with the gene engineering bacterial, the problems of low yield, more by-products, long period, high energy consumption and the like of fermentation of the gamma-polyglutamic acid gene engineering bacterial under high viscosity and dissolved oxygen limiting conditions are solved, and the gamma-polyglutamic acid gene engineering bacterial can be applied for large-scale industrial production of gamma-polyglutamate acid.

The invention discloses gene recombination bacillus amyloliquefaciens and a microbial inoculum preparation method. A bacillus amyloliquefaciens FZB42 strain is taken as a host bacterium to construct the gene recombination bacillus amyloliquefaciens XLV09-1 with transparent vitreoscilla hemoglobin which is regulated and controlled by a P43 strong promoter by contrasting a gene recombination vector. The produced spore number of microbial inoculum prepared by fermenting the bacillus amyloliquefaciens can reach more than 100 hundred millions per milliliter, the overall yield of antibacterial lipopeptid reaches more than 1 gram per liter and prevention effect on various fungal plant diseases is improved by over 30 percent. The microbial inoculum is a green microecological preparation, of no toxic residue, safe to human and livestock, and favorable for protecting the ecological environment.

The invention discloses a recombinant plasmid for expressing vitreoscilla hemoglobin in a cell and a construction method of the recombinant plasmid, a genetic engineering bacterium of Gluconobacter oxydans (G.oxydans) and a construction method and application of the genetic engineering bacterium. The recombinant plasmid comprises a vitreoscilla hemoglobin gene vgb, a PtufB promoter of the Gluconobacter oxydans (G.oxydans) and a suitable carrier, wherein the genetic engineering bacterium is obtained by transforming recombinant plasmid into a host bacterium. The genetic engineering bacterium can be used for expressing the vitreoscilla hemoglobin in the cell, can be used for improving the yield of biomass and catalysate 1, 3-dihydroxyacetone under the condition of not changing the existing equipment and energy consumption pressure and provides an effective path for solving the problem of in sufficient oxygen supply in the cell cultivation and catalytic process of the Gluconobacter oxydans (G.oxydans).

The invention relates to a bacillus licheniformis with a vitreoscilla hemoglobin gene. The bacterial strain is the bacillus licheniformis (PHY300-vgb), wherein the bacterial strain is stored in China Typical Model Cultivation Center of Wuhan, and the storage number is CCTCC NO: M2011345. The invention aims to provide the bacterial strain of the bacillus licheniformis with vitreoscilla hemoglobin gene, which can produce bacitracin in high yield, as well as a construction method and an application of the bacterial strain.

The invention provides a method for constructing a high-yield gamma-polyglutamic acid engineering strain by using gene engineering technology, and aims to greatly improve the yield of the gamma-polyglutamic acid at a lower dissolved oxygen level. In the method of the invention, a hemoglobin gene (vgb) of vitreoscilla hemoglobin is imported into a gamma-polyglutamic acid producing strain cell by using the gene engineering technology, so that the vitreoscilla hemoglobin (VHb) is successfully expressed; and thus, the utilization ratio of oxygen by a recombinant strain and the yield of producing the gamma-polyglutamic acid by a fermentation method are improved. Results show that the capacity of producing the gamma-polyglutamic acid by the recombinant strain is improved by 1.8 times during the fermentation, so that the problem of high dissolved oxygen requirement during the large-scale production of the gamma-polyglutamic acid is solved, and the feasibility is provided for the large-scale industrial production of the gamma-polyglutamic acid.

The invention relates to a vitreoscilla hemoglobin vgbS nucleotide sequence and a plasmid and a preparation method thereof, and belongs to the technical field of biological engineering. The vitreoscilla hemoglobin vgbS nucleotide sequence is prepared by introducing restriction enzyme cutting sites of NdeI and EcoRI at both ends of a vgb nucleotide sequence of streptomyces serving as a host cell respectively. In addition, the plasmid pJTU4406 is generated by cutting off the vgbS nucleotide sequence on pJTU4405 through the NdeI and the EcoRI, and inserting the vgbS nucleotide sequence into corresponding sites of a carrier pIB139. An amino acid sequence of protein expressed and generated by the vgbS nucleotide sequence is totally identical to that of the natural vitreoscilla hemoglobin, and the vitreoscilla hemoglobin vgbS nucleotide sequence can improve the utilization efficiency of the streptomyces to oxygen, promote the growth velocity of thalli and improve the yield of antibiotics.

The invention discloses recombination bacillus subtilis of a vitreoscilla hemoglobin mutant gene with optimized controllable expression codon and and application thereof. A strain is the recombination bacillus subtilis/PBE-Pxyl-vgb, and the collection number is CGMCC No.10787. A bacillus subtilis expression carrier PBE-Pxyl-vgb is constructed through the vitreoscilla hemoglobin mutant gene vgh with the optimized codon; the expression carrier PBE-Pxyl-vgb is electrically transferred into bacillus subtilis, and the recombination bacillus subtilis (PBE-Pxyl-vgb) is obtained. When intermediate temperature alpha-amylase is produced with the strain, xylose is added or not added according to the requirements in the practical fermentation process to start or stop the expression of vitreoscilla hemoglobin, the controllable expression of the vitreoscilla hemoglobin gene is achieved, excessive consumption, caused by the continuous expression of the vitreoscilla hemoglobin, of nutrient substances in fermentation liquor and oxygen is avoided, and therefore the oxygen use efficiency of the recombination bacillus subtilis is improved. The result shows that the intermediate temperature alpha-amylase fermentation level of the recombination bacillus subtilis is increased by 303% compared with an original strain.

The invention discloses recombinant streptomyces nodosus capable of producing amphotericin and applications of the recombinant streptomyces nodosus. The recombinant streptomyces nodosus is obtained byimporting vitreoscilla hemoglobin (Vhb), S-adenosylmethionine synthetase (Metk), amphotericin synthetic gene cluster regulating factor (AmphRIV), secondary metabolite global regulation factor (AraC)and erythrocin strong promoter ermE*p sequences as shown in SEQ ID NO.1, SEQ ID NO.2, SEQ ID NO.3, SEQ ID NO.4 and SEQ ID NO.5 into streptomyces nodosus ZJB2016050. Through importing and overexpression of the gene, in recombinant streptomyces nodosus, the yield of amphotericin B is improved by about 45%, the yield of the by-product amphotericin A is reduced by 60%, meanwhile, the growth cycle of the thallus during the fermentation process is shortened, and thus the purpose of improving the production efficiency is achieved.

The invention discloses a ganoderic acid high-producing engineering strain kmust-VGB-1, which is preserved in China General Microbiological Culture Collection Center with preservation number of CGMCC NO. 11301. The lucid ganoderma engineering strain kmust-VGB-1 for high production of ganoderic acid is prepared by expressing a vitreoscilla hemoglobin VGB gene through a lucid ganoderma strong promoter P-gpd; through a fermentation experiment in a shaking flask, under the circumstance of avoiding influence on cell growth, yield of monomer ganoderic acids GA-Me, GA-T, GA-Mk and GA-S produced by the VGB converter strain are respectively 3.1 times, 3.2 times, 2.1 times and 3.6 times that of monomer ganoderic acids produced by a WT strain; therefore, the high-producing strain, as an engineering strain for producing the ganoderic acid, has a broad application prospect.

The invention relates to a poor oxygen resistance high density fermentation natamycin gene engineering strain (Streptomyces gilvosporeus sw-807, CGMCC No.6890) constructed by using vgb gene, and an application thereof. According to the present invention, vitreoscilla hemoglobin gene vgb is adopted to construct recombinant plasmid, a conjugal transfer manner is adopted to integrate the vgb gene into Streptomyces gilvosporeus genome to obtain the recombinant gene engineering strain capable of being directly used for natamycin fermentation production, and VHb expression in the recombinant strain can be provided for effectively solving oxygen supply and demand contradiction in natamycin fermentation production so as to substantially increase natamycin yield, reduce production cost and bring great economic benefits.

The invention relates to the field of genetic engineering, and discloses a vitreoscilla hemoglobin gene expression box and a method for improving yield of saccharifying enzyme produced by aspergillus niger. The expression box comprises a transparent vitreoscilla hemoglobin gene, and a promoter and a terminator of a glyceraldehyde-3-phosphate dehydrogenase gene. In the invention, aspergillus brasiliensis is subject to molecular modification, the problem of oxygen depletion of the aspergillus niger under the oxygen-limiting condition is solved from the molecular level and the energy consumption is reduced in a process of producing the saccharifying enzyme by the deep fermentation of the aspergillus niger liquid.

The present invention discloses mutational Vitreoscilla hemoglobin gene sequence and its coded Vitreoscilla hemoglobin mutant, as well as the transformant obtained through transforming heterogenous host with the mutational Vitreoscilla hemoglobin gene and its application in fermentation. The mutational Vitreoscilla hemoglobin gene consists of 438 nucleotides and codes one segment of protein comprising 146 amino acids. The gene expressed protein can raise the biomass of the host cell in the condition of insufficient environmental oxygen dissolution, so that it may be applied for promoting the growth of the host cell in fermentation industry to raise the yield of the target product and lower production cost.

The invention relates to a tylosin gene engineering strain. The construction method for the tylosin gene engineering strain comprises the following steps: cloning tylosin through polymerase chain reaction (PCR) to synthesize crucial rate-limiting enzyme gene TylF and Vitreoscilla hemoglobin VHb gene, constructing a recombinant plasmid by using an E. coli-streptomyces shuttle vector, importing the recombinant plasmid into Escherichia coli ET12567/pUZ8002, performing conjugal transfer culture on the Escherichia coli ET12567/pUZ8002 and streptomyces fradiae spores, converting streptomyces, screening a converter through apramycin resistance, further confirming the converter by using PCR, and finally obtaining the gene engineering strain. The tylosin gene engineering strain can be directly used in the industrial production of tylosin; and compared with the tylosin gene engineering strain used in the conventional industrial production, the tylosin gene engineering strain has the advantages that the yield of the tylosin can be obviously improved to be 17,000-18,000u/ml and the content of a component A in the tylosin is effectively improved to be 94-95 percent.

The invention discloses a method for improving the moisture resistance of cabbage type rape. The method comprises the following steps of: (A), constructing a glufosinate-ammonium resistant expression vector containing vitreoscilla hemoglobin (vgb), namely, (1) obtaining a target fragment, (2) recovering a polymerase chain reaction (PCR) product, transforming and sequencing, (3) preparing transferred-deoxyribonucleic acid (T-DNA), (4) preparing Vector, (5) preparing the expression vector, and (6) preparing bacterial liquid of agrobacterium tumefaciens; (B), breeding a recovery line of the moisture-resistant cabbage type rape, namely transforming the vgb into R2 by an agrobacterium tumefaciens mediated method to obtain a T0 recovery line, and performing continuous selfing to obtain a T2 recovery line; (C), breeding a maintainer line of the moisture-resistant cabbage type rape, namely (a) transforming the vgb into 6098B by the agrobacterium mediated method to obtain a T0 maintainer line, and performing continuous selfing to obtain a T2 maintainer line, and (b) hybridizing a T1 recovery line single plant serving as a male parent with the 6098B, performing continuous backcrossing by taking the 6098B as a recurrent parent to obtain BC3, and performing selfing to obtain the T2 maintainer line; and (D), obtaining a sterile line of the moisture-resistant cabbage type rape. The method is easy and convenient to operate and obvious in effect, the cost can be reduced, and the breeding progress can be shortened.

The invention discloses a method for improving sorangium cellulosum's epothilone B yield by genetic engineering. The method includes: introducing a Vitreoscilla hemoglobin gene vgb into an expression vector PBEP43 to obtain a recombinant plasmid PBEP43-vgb, and then subjecting the recombinant plasmid PBEP43-vgb to electrotransformation into sorangium cellulosum So ce M4, thus obtaining a sorangium cellulosum So ce M4 recombinant strain, and realizing improving the sorangium cellulosum So ce M4's epothilone B yield. According to the invention, the electrotransformation method is employed to transform the gene of sorangium cellulosum So ce M4, thereby greatly improving the yield of epothilone B. The method provided by the invention has broad application prospects, and research in the aspect is not reported to date.

The invention relates to an artificially-synthesized vitreoscilla hemoglobin gene. The nucleotide sequence of the gene is shown in SEQ ID No.1, and the gene is marked as vgbL. The invention further provides a corresponding genetically-engineered strain for producing enramycin. The invention further provides a method for constructing the genetically-engineered strain for producing the enramycin. The artificially-synthesized vitreoscilla hemoglobin gene, the strain and the method have the advantages that the nucleotide sequence of the novel vitreoscilla hemoglobin gene vgbL which can be efficiently expressed in streptomyces is obtained, and the gene is artificially synthesized and transferred into fungicidal streptomyces ATCC21013; compared with an original wild strain, by means of the genetically-engineered strain, the yield of the enramycin is increased by 16.6%, and therefore, it can be seen that the vgbL can become an effective means of increasing the yield of antibiotics in variousstreptomyces.

The invention discloses a yeast engineering bacterial strain capable of producing glutathione, which has the bacterial strain name of 101-V and the classification name of Saccharomyces cerevisiae strain collected in the China General Microbiological Culture Collection Center on February 15, 2012, with the collection number of CGMCC NO. 5758. The invention also discloses a method for establishing an engineering bacterial strain capable of producing glutathione at a high yield. The method comprises the following steps: integrating a Vitreoscilla hemoglobin gene vgb into a yeast genome capable of producing glutathione, and screening to obtain a recombinant yeast capable of expressing transparent Vitreoscilla hemoglobin gene. Therefore, the oxygen utilizing capability of the hemoglobin gene recombinant strainis improved in a molecular level, the contradiction between supply and demand of oxygen in the microbiological fermenting process is solved, the consumption of oxygen and energy is lowered, and the yield of glutathione is increased.

The invention belongs to the field of microbial engineering, provides a streptomyces clavuligerus strain LNSC with low oxygen consumption and high fermentation yield, a strain preparation method and application of the strain, and particularly relates to a method. The method utilizes genetic engineering methods, researches the application aspect of vitreoscilla hemoglobin gene (vhb), transfers the gene into the streptomyces clavuligerus culture that is used for clavulanic acid production, obtains high expression of VHB protein, thus improving the production characteristics of the streptomyces clavuligerus, and lowering the energy consumption of zymolytic production.

The invention discloses a gene-engineered bacterium of gene-recombined peoriae paenibacillus, and a construction method and application of the gene-engineered bacterium. Gene-recombined peoriae paenibacillus BC39 with vitreoscilla hemoglobin regulated and controlled by a P43 strong promoter is constructed by constructing a gene-recombination integrative vector with a peoriae paenibacillus LXH-B-1 strain as host bacteria. The gene-engineered bacterium of the gene-recombined peoriae paenibacillus is capable of intracellularly expressing the vitreoscilla hemoglobin, so that the utilization rate of oxygen by the peoriae paenibacillus, the cell biomass liveweight and the yield of catalysate polypeptide antibiotics can be improved; and the yield of the catalysate polypeptide antibiotics of the engineering strain is increased by 45% compared with that of the original strain; and therefore, an effective way is provided for solving the problem of insufficient oxygen supply in the cell culture and catalysis process of the peoriae paenibacillus.

The invention discloses self-organized recombinant vitreoscilla hemoglobin with self-organizing ability and application of a gene thereof in gene engineering product production and belongs to the technical field of gene engineering. The self-organized recombinant vitreoscilla hemoglobin is composed of a vitreoscilla hemoglobin module, a connecting peptide module and a self-organized module, amino acid sequence of the vitreoscilla hemoglobin module is SEQ ID NO.1, amino acid sequence of the connecting peptide module is (SEQ ID NO.2)n, n value refers to number of series-connection repeating times of the connecting peptide module, and n is an integer from 2 to 10; amino acid sequence of the self-organized module is SEQ ID NO.3 or SEQ ID NO.4. Compared with vitreoscilla hemoglobin, protein expressed by the gene has self-organizing ability, the self-organized recombinant vitreoscilla hemoglobin has higher plasmalemma combining performance, biomass and gene engineering product yield of host cells can be increased under the circumstance that dissolved oxygen in environment is insufficient, and production cost is lowered.