429results about "Mutant preparation" patented technology

Some aspects of this disclosure provide strategies, systems, reagents, methods, and kits that are useful for the targeted editing of nucleic acids, including editing a single site within the genome of a cell or subject, e.g., within the human genome. In some embodiments, fusion proteins of Cas9 and nucleic acid editing enzymes or enzyme domains, e.g., deaminase domains, are provided. In some embodiments, methods for targeted nucleic acid editing are provided. In some embodiments, reagents and kits for the generation of targeted nucleic acid editing proteins, e.g., fusion proteins of Cas9 and nucleic acid editing enzymes or domains, are provided.

Strategies, systems, methods, reagents, and kits for phage-assisted continuous evolution are provided herein. These include strategies, systems, methods, reagents, and kits allowing for stringency modulation to evolve weakly active or inactive biomolecule variants, negative selection of undesired properties, and / or positive selection of desired properties.

The present disclosure relates to enriched engineered glycoprotein populations with a therapeutically optimized heterogeneous glycan profile. The heterogeneous glycan profile comprises sialylated complex type glycans, which are associated with improved efficacy of the glycoprotein. The present disclosure also relates to compositions and uses of the engineered glycoprotein populations and cell-based method of producing them.

A microorganism according to the present disclosure is a microorganism (accession number: NITE BP-03832) belonging to Bacillus velezensis, a microorganism (accession number: NITE BP-04073) belonging to Bacillus velezensis, a microorganism (accession number: NITE BP-04126) belonging to Bacillus velezensis, or a variant of these having a plant disease prevention effect.

A Schizochytrium algae strain rich in n-3 fatty acid and a screening and culture method therefor. Compared with a conventional strain, the content of EPA and the content of n-3 DPA in the provided strain are greatly improved, and the strain has good genetic stability, fast growth rate and good industrialization prospects. Also provided are a culture medium for mutagenesis of Schizochytrium sp., a culture medium for screening Schizochytrium sp., and a suitable mutagenesis method and screening method.

The invention provides a phaffia rhodozyma mutant strain for high yield of astaxanthin as well as a screening method and application of the phaffia rhodozyma mutant strain, belongs to the technical field of fermentation microorganism screening, and can solve the problems of low astaxanthin yield, high cost and low growth temperature of wild type phaffia rhodozyma existing in synthesis of astaxanthin from a phaffia rhodozyma strain. The invention provides a phaffia rhodozyma mutant strain capable of producing astaxanthin at high yield, the phaffia rhodozyma mutant strain is a phaffia rhodozyma AA0529 mutant strain, the phaffia rhodozyma AA0529 mutant strain is preserved in China Center for Type Culture Collection in Wuhan, Hubei on September 7, 2023, and the preservation number is CCTCC (China Center for Type Culture Collection) NO: M20231637. According to the method, the yield of the astaxanthin can be increased, and the yield of the astaxanthin is increased to 3.3 mg / g at 26 DEG C.

The invention discloses a method for screening high-yield recombinant human lactoferrin through nucleic acid aptamer-assisted mutagenesis of a trichoderma reesei strain and application, and belongs to the technical field of microorganisms. The nucleic acid aptamer Lac-6a-FAM-BHQ1 capable of changing the fluorescence intensity according to the content of lactoferrin is constructed on the basis of the aptamer Lac-6a, and the nucleic acid aptamer can be used for rapidly selecting a mutant strain for producing high-titer recombinant human lactoferrin. The ARTP mutagenesis technology is combined with a nucleic acid aptamer auxiliary screening system, the mutant strain with the improved recombinant human lactoferrin yield is rapidly obtained from a large number of mutant strains, and the yield reaches 274.57 mg / L and is improved by 24.8% compared with that of an original strain. After the genes 111681 and 62716 are over-expressed in T. reesei, compared with a control strain, the yields of the recombinant human lactoferrin of the recombinant strain are respectively increased by 6.9% and 22.1%.

In traditional plant breeding approaches, chemical mutagenesis may be utilized to introduce nucleotide substitutions at random in the genome of a plant, i.e. without possibilities to control the sites of nucleotide changes. Because of genome complexities, the statistical probability is extremely little when it comes to finding a predetermined nucleotide substitution. The present invention, however, demonstrates how a novel, alternative use of digital polymerase chain reaction (dPCR), preferably droplet dPCR (ddPCR), is developed to exploit finding of specific nucleotide substitutions in mutated genes. The entire platform comprises a screening method with a library of mutagenized organisms, digital PCR-based systems and a set-up to propagate and analyze identified, mutated organisms.

Certain one or more embodiments of the instant disclosure are related to, inter alia, recombinant (genetically modified) filamentous fungal cells (strains) producing proteins of interest, methods and compositions for the design and construction of modified filamentous fungal cells producing proteins of interest, methods and compositions for the expression / production / secretion / recovery and the like of endogenous and / or heterologous proteins of interest (e.g., phytases, lipases, glucoamylases, phospholipases, esterases, cellulases, hemicellulases, xylanases, etc.) in modified filamentous fungal cells, methods and compositions for producing proteins of interest in recombinant filamentous fungal, wherein the proteins produced and secreted into the broth and / or proteins recovered from the broth have uniform and consistent N-linked glycosylation patterns, and / or reduced (unwanted) glycation of one or more proteins of interest, and the like.

The invention discloses a fermentation process for shortening a fermentation period and efficiently synthesizing Ectoine. The fermentation process comprises the following steps: screening and activating high-yield strains, preparing a fermentation culture medium, performing fermentation culture, performing sectional dissolved oxygen regulation and control, performing double-stage pH dynamic regulation and control, performing carbon-nitrogen ratio dynamic regulation based on metabolic flow analysis, performing osmotic pressure gradient induction in the later stage of fermentation, and extracting and purifying a product. According to the fermentation process for shortening the fermentation period and efficiently synthesizing the Ectoin, the expression quantity of key enzymes for synthesizing the Ectoin is increased by 80% or above compared with that of an original strain through the Halomonassp.ECT-09 strain bred through ultraviolet-nitrosoguanidine compound mutation, the basic yield reaches 35 g / L, a foundation is laid for high yield, and the method is suitable for industrial production. A glucose-trehalose mixed carbon source and a yeast extract-ammonium nitrate mixed nitrogen source are adopted, and an L-aspartic acid-L-asparagine mixed precursor is added, so that the metabolic pathway is shortened, and the loss of an intermediate product is reduced.

The invention provides Saccharomyces cerevisiae for producing squalene as well as a screening method and application of the Saccharomyces cerevisiae. The method comprises the following steps: culturing a Saccharomyces cerevisiae engineering strain ySC782 for synthesizing squalene to obtain a to-be-mutagenized bacterial solution, carrying out ARTP mutagenesis on the to-be-mutagenized bacterial solution, adding the to-be-mutagenized bacterial solution into a YPD liquid culture medium, culturing to obtain a resuscitation bacterial solution, adding the resuscitation bacterial solution into a sucrose solution with gradient concentration distribution, screening, and repeating the ARTP mutagenesis-sucrose solution screening step to obtain the squalene mutant strain. And finally, the saccharomyces cerevisiae strain ySC782-M5 is obtained through screening, and the saccharomyces cerevisiae strain ySC782-M5 is obtained through screening. The yield of squalene produced by the saccharomyces cerevisiae ySC782-M5 reaches 49.83 g / L, so that a solution is provided for breaking through the yield bottleneck of squalene synthesized by yeast.

To provide methods for mutating wild type Saccharomyces cerevisiae yeast to enhance the ability to produce both glutathione and aldehyde dehydrogenase.SOLUTION: The invention relates to a yeast strain that produces glutathione (GSH) and an aldehyde dehydrogenase (acetaldehyde dehydrogenase). More specifically, the invention relates to Saccharomyces cerevisiae Kwon P-1 KCTC13925BP, Saccharomyces cerevisiae Kwon P-2 KCTC14122BP and Saccharomyces cerevisiae Kwon P-3 KCTC14123BP that produce both glutathione and an aldehyde dehydrogenase.SELECTED DRAWING: Figure 6

The invention belongs to the technical field of microorganisms, and particularly relates to Chryseobacterium sp. 2R14A and application of the Chryseobacterium sp. 2R14A. The strain is excellent in oxidation resistance, still can grow after being treated by 80mM hydrogen peroxide for 30 minutes, and is not completely inactivated after being treated by 160mM hydrogen peroxide, and a genome contains antioxidant related genes; growth conditions are mild, sodium chloride is not needed, large-scale culture is easy, cost is low, and physiological and biochemical characteristics are stable. The strain can be used for treating oxidizing agent-containing polluted environment and improving stress resistance of crops or microorganisms, can also be used as a gene engineering recipient bacterium or a mutation breeding material, and expands application in the fields of agriculture, environment and medicine.

The invention provides a microorganism as well as an application and a method of the microorganism in preparation of 1, 3-propylene glycol and lactic acid, the microorganism is lactobacillus reuteri, the microorganism is preserved in Guangdong Microbial Culture Collection Center on May 29, 2024, and the preservation number is GDMCC NO: 64700. The invention further provides a method for preparing 1, 3-propylene glycol and lactic acid, and the microorganism is lactobacillus reuteri and is preserved in Guangdong Microbial Culture Collection Center on May 29, 2024. The Lactobacillus reuteri can co-produce 1, 3-propylene glycol and lactic acid, has the advantages of high yield of 1, 3-propylene glycol and lactic acid, fast growth and metabolism, strong stability, high safety and the like, provides a new idea for green, low-cost and efficient production of 1, 3-propylene glycol and lactic acid, and has a wide application prospect.

To provide an artificial gene and a gene mutation method which are not subject to gene recombination regulations and enable the introduction of a local mutation into a target gene and enable the quantitation of the mutation at the time of mutation introduction.SOLUTION: An artificial gene has a 15N abundance ratio exceeding a natural abundance ratio in bases of at least a part of DNA. A gene mutation method includes: a first step of producing a state in which 15N is unevenly distributed in a prescribed DNA in a living cell; and a second step of irradiation with a proton beam at an energy at which 15N produces a resonant nuclear reaction.SELECTED DRAWING: Figure 1

The invention belongs to the field of microbial breeding and fermentation engineering, and particularly relates to a saccharomyces cerevisiae mutagenesis strain with high yield of S-adenosyl-L-methionine as well as a mutagenesis method and application of the saccharomyces cerevisiae mutagenesis strain. Saccharomyces cerevisiae is induced to generate genetic diversity through multiple rounds of ultraviolet mutagenesis, and high-throughput screening is performed in combination with strain color phenotypic difference; and the mutant strain HY2402-Z18 with high yield of S-adenosine-L-methionine and genetic stability is obtained. The SAM yield of the mutagenic strain is increased by 40.1% compared with that of an original strain, the SAM yield reaches 13.44 g / L when the mutagenic strain is fermented for 60 h in a 5 L fermentation tank system, the unit yield reaches 0.156 g / L / OD, and the methionine conversion rate is 75%. After thalli are collected through centrifugation of fermentation liquor, an S-adenosine-L-methionine product with the purity of 98%-100% can be obtained through purification of ion exchange resin, no by-products are accumulated, the yield, the substrate conversion rate and the production efficiency of the S-adenosine-L-methionine can be remarkably improved when the mutagenesis strain is used for fermentation production, and the production cost is reduced. The production cost is effectively reduced, the product purity and quality are improved, and the method is suitable for industrial large-scale production of SAM.

Certain one or more embodiments of the present disclosure are directed, inter alia, to recombinant (genetically modified) filamentous fungal cells (strains) that produce proteins of interest, methods and compositions for designing and constructing modified filamentous fungal cells that produce proteins of interest, for endogenous and / or heterologous proteins of interest (in modified filamentous fungal cells). The present invention relates to methods and compositions for expression / production / secretion / recovery and the like of recombinant filamentous fungi (e.g., phytase, lipase, glucoamylase, phospholipase, esterase, cellulase, hemicellulase, xylanase and the like), methods and compositions for the production of proteins of interest in recombinant filamentous fungi, and methods and compositions for the production of proteins of interest in recombinant filamentous fungi. Wherein the proteins produced and secreted into and / or recovered from the bouillon have a homogeneous and consistent N-linked glycosylation pattern, and / or a (unwanted) reduction in saccharification of one or more proteins of interest, and the like.

The application discloses a Yarrowia lipolytica strain with high yield of gamma-decalactone and / or delta-decalactone and application thereof, and belongs to the technical field of biotechnology. The name of the strain is Yarrowia lipolytica Y-1, and the preservation number is GDMCC No: 67810. The strain Y-1 can be directly used for biological transformation to cooperatively and efficiently prepare gamma-decalactone and delta-decalactone with castor oil acid as a substrate. The strain has three outstanding advantages: ① yield advantage; ② tolerance advantage; and ③ application advantage. The biological transformation method has the advantages of mild reaction condition, low cost, product meeting natural perfume standards, environmental friendliness and the like.

The invention relates to a gene expression cassette and application thereof. The gene expression cassette comprises a tyrosinase gene, a transcriptional repressor protein gene, a melanin response promoter and a reporter gene, and the reporter gene is located at the downstream of the melanin response promoter and is regulated by the promoter. The gene expression cassette with a specific structure is designed, melanin in cells can be responded, a melanin biosensor can be further constructed by utilizing the gene expression cassette, the gene expression cassette has good specificity, and high-throughput screening of strains can be realized. Furthermore, ALE evolution is carried out on the strain for producing melanin, high-throughput screening is carried out based on a designed sensor, the strain for producing melanin with high yield is obtained, through shake-flask culture and 5L fermentation tank culture, the yield of melanin reaches up to 10.84 g / L and 29.04 g / L respectively, and an efficient and feasible solution is provided for large-scale production of melanin.

To provide methods for culturing cyanobacteria.SOLUTION: A method for cultivating cyanobacteria includes: providing the cyanobacteria; subjecting the cyanobacteria to physical mutagenesis, so as to obtain primary mutant cyanobacteria; subjecting the primary mutant cyanobacteria to chemical mutagenesis, so as to obtain secondary mutant cyanobacteria; and subjecting the secondary mutant cyanobacteria to a temperature resistance test and an environment resistance test, so as to obtain target cyanobacteria, where the lethal rate of the physical mutagenesis ranges between 50% and 80%, and the lethal rate of the chemical mutagenesis ranges between 40% and 55%.SELECTED DRAWING: Figure 1

This invention relates to a hangover relief composition containing a dried powder, lysate, or extract of enzymes that produce glutathione and aldehyde dehydrogenase. Specifically, this invention relates to a hangover relief composition containing dried powder, lysate, or extract of *Saccharomyces cerevisiae* Kwon P-1 KCTC 13925BP, *Saccharomyces cerevisiae* Kwon P-2 KCTC14122BP, and *Saccharomyces cerevisiae* Kwon P-3 KCTC14123BP, which simultaneously produce glutathione and aldehyde dehydrogenase.

The present invention relates to yeast strains of the species Saccharomyces pastorianus that have the useful property of producing low levels of diacetyl during fermentation. Methods for producing malt- and / or grain-based beverages using these strains, as well as the beverages produced thereby, are also provided.

Provided is a tobacco product having improved fragrance and flavor. Specifically provided is a tobacco material including part of a plant of the genus Nicotiana, wherein: the plant of the genus Nicotiana has a mutation associated with the suppression of a function of an endogenous CAD gene; and the tobacco material contains 0.05 ppm or more of vanillin.

A method for producing a fermented milk product (e.g. a yogurt) with a relatively high stable pH value at the end of the fermentation comprising inoculating milk with Streptococcus thermophilus (ST) Gal(+) bacteria.

The invention discloses chlorella capable of resisting acetate and urea and a breeding method and application thereof, and relates to the technical field of microorganisms, the chlorella is a Sorokiniana mutant strain, the classification name of the chlorella is Chlorella sorokiniana-TongWei-01, the chlorella is preserved in the China General Microbiological Culture Collection Center at 2025-12-1, and the preservation number of the chlorella is CGMCC NO. The address of the preservation unit is Institute of Microbiology, Chinese Academy of Sciences, No.3, No.1 Yard, Beichen West Road, Chaoyang District, Beijing, and the preservation number is CGMCC No.46630. According to the method, large-scale culture can be performed by taking acetic acid / sodium acetate as a unique carbon source and urea as a unique nitrogen source, the method has excellent acetic acid utilization efficiency and urea tolerance, toxicity interference is resisted while two substrates are efficiently converted, high-density culture supported by a cheap carbon-nitrogen source is realized, and a core algae species support is provided for large-scale production of microalgae.

Some aspects of this disclosure provide compositions, methods, systems, and kits for controlling the activity and / or improving the specificity of RNA-programmable endonucleases, such as Cas9. For example, provided are guide RNAs (gRNAs) that are engineered to exist in an “on” or “off” state, which control the binding and hence cleavage activity of RNA-programmable endonucleases. Some aspects of this disclosure provide mRNA-sensing gRNAs that modulate the activity of RNA-programmable endonucleases based on the presence or absence of a target mRNA. Some aspects of this disclosure provide gRNAs that modulate the activity of an RNA-programmable endonuclease based on the presence or absence of an extended DNA (xDNA).

The invention relates to methods of producing a baked product, said method comprising adding a mature thermostable variant of a parent glucoamylase at least 70% identical to SEQ ID NO:1, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8 or SEQ ID NO:10 to a dough, adding less than 150% (baker's %) of sugar to the dough, and baking the dough as well of uses of a mature 5 thermostable variant of a parent glucoamylase at least 70% identical to SEQ ID NO:1, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8 or SEQ ID NO:10 for producing a baked product, wherein the variant is added to a dough, less than 150% (baker's %) of sugar or sucrose is added to the dough, and the dough is baked. 10

The invention relates to a high-yield protein glutaminase Chryseobacterium prion strain and a preparation method thereof. The strain is obtained by carrying out ARTP and NTG compound mutagenesis treatment on an original strain of Chryseobacterium prion and carrying out high-throughput screening on the strain subjected to the compound mutagenesis treatment. The strain is preserved in the China Center for Type Culture Collection, and the preservation number is CCTCC M 20251792. The invention also relates to a method for preparing protein glutaminase by using the strain of the invention. The enzyme activity of the protein glutaminase of the strain is 6 times that of an original strain, and the production efficiency of the protein glutaminase can be remarkably improved.

The invention relates to the technical field of microbial fermentation and food processing, and particularly discloses Weissella fusiformis JMF-006 as well as a mutagenesis method and application thereof. The invention provides a Weissella fusiformis JMF-006 with the exopolysaccharide yield of 20-25g / L. The Weissella fusiformis JMF-006 is obtained by breeding through a mutation breeding method of a high-yield EPS strain, and the mutation breeding method can also be used for culturing other high-yield EPS strains. The invention provides a strain composition, the strain composition comprises Weissella fusiformis JMF-006 or a high-yield EPS strain, other lactic acid bacteria strains (such as lactobacillus plantarum JMF-001 and lactobacillus casei JMF-002) can be further compounded, and the obtained strain composition has more excellent fermentation performance. When the Weissella fusiformis JMF-006, the high-yield EPS strain and the strain composition provided by the invention are applied to the fermentation process of food-derived drinks, a fermented product with stable properties and better flavor can be obtained, and the technical problems that the yield of extracellular polymeric substances is low, the fermentation performance is poor and the application range is narrow in an existing lactic acid bacteria starter are solved.

The invention discloses a method for preventing and treating sclerotinia rot of colza by antagonistic trichoderma strains, and relates to the technical field of agricultural disease prevention and control, and the method comprises the following specific steps: S100, strain screening and mutagenesis: separating trichoderma strains from rhizosphere soil of colza in different environments, primarily screening strains with antagonistic action on sclerotinia sclerotiorum by a plate confrontation method, and carrying out mutagenesis on the strains with antagonistic action on sclerotinia sclerotiorum; carrying out chemical mutagenesis treatment on the primarily screened strains, carrying out confrontation culture on the primarily screened strains and sclerotinia sclerotiorum again, screening out high-yield antagonistic substances with larger inhibition zones and mutant strains with strong stress resistance, and culturing to obtain trichoderma fermentation liquor; by obtaining the trichoderma strains which can keep efficient antagonism and strong stress resistance under different environmental conditions, the stability of the trichoderma strains on disease control is enhanced, so that the rape can effectively resist invasion of sclerotiniose under different environmental conditions in the growth process, and the disease resistance of the rape is improved. And antagonistic active substances are extracted to prepare a biological fertilizer carrier, and the biological fertilizer carrier is applied to the planting process of oilseed rape, so that the growth and reproduction of sclerotinia sclerotiorum are effectively inhibited.