128results about "Lyases" patented technology

This invention relates to the field of low-temperature fermentation formulation technology. The invention provides a low-temperature resistant compound bacterial enzyme formulation, its preparation method, and its application in the co-fermentation of straw and manure. The compound bacterial enzyme formulation comprises the following raw materials in parts by weight: 8-10 parts of compound bacterial agent, 1-1.4 parts of compound enzyme agent, and 88.6-91 parts of excipients. The compound bacterial enzyme formulation of this invention can ferment straw and manure under conventional conditions as well as at low temperatures of around 15°C, effectively meeting the fermentation requirements in low-temperature regions. By combining the excipients with the bacteria and enzymes, substances and nutrients during the fermentation process can be effectively adsorbed, avoiding acid inhibition, and providing attachment sites for the bacteria and enzymes, thus prolonging their activity.

This invention relates to the fields of bioengineering and fermentation engineering, and discloses a genetically engineered *Streptomyces albopictus* strain that produces high levels of ε-polylysine and its fermentation production process. This genetically engineered strain uses *Streptomyces albopictus* strains acclimated to ε-polylysine tolerance as a chassis, and its genome integrates... ppc and dapf Through expression box, heterogeneous asd Expression cassettes, containing secretory signal peptides lysp Fusion expression cassettes and D404A / K499A point mutations pls Expression cassette. The fermentation process employs a two-stage pH control strategy, maintaining a neutral environment during the cell growth phase and adjusting to an acidic environment during product synthesis. This invention improves ε-polylysine yield through a synergistic strategy of enhancing precursor supply, constructing efflux detoxification channels, and increasing synthase activity; combined with an acidic fermentation process, it effectively inhibits product degradation, yielding a high-molecular-weight and highly uniform target product suitable for industrial production.

The application belongs to the technical field of bioengineering, and particularly relates to a xanthan gum cleavage ancestral enzyme AncXLY196 based on ancestral sequence reconstruction, an amino acid sequence of which is shown as SEQ ID NO. 2. Based on the xanthan gum cleavage ancestral enzyme, a mutant AncXLY196-X7 is further obtained, an amino acid sequence of which is shown as SEQ ID NO. 4, xanthan gum degradation activity of which reaches 2.5 times of that of the xanthan gum cleavage ancestral enzyme, and the optimal catalytic temperature of the xanthan gum cleavage ancestral enzyme and the mutant thereof is increased to 45 DEG C, and the enzyme activity of the mutant of the xanthan gum cleavage ancestral enzyme is reserved by 50-80% after being treated at 45-55 DEG C for 12-24 h. The application provides a new enzyme resource for efficient enzymatic degradation of xanthan gum and preparation of functional oligosaccharides.

Provided herein are methods amenable to high-throughput multiplexing, in part, using a modified enzyme complementation assay, that can be used to screen a library of test compounds and to identify compounds that inhibit denaturation of a target polypeptide of interest.

This invention relates to a cytochrome P450 oxidase STO1, its encoding gene, and its applications. This invention is the first to clone and identify a cytochrome P450 oxidase gene, STO1, from potato responsible for catalyzing the synthesis of spirogelothoraxone, and to determine the nucleotide sequence and translated amino acid sequence of this gene. This invention provides expression vectors, recombinant microorganisms, or transgenic cell lines containing this gene. This invention constructs an engineered Saccharomyces cerevisiae strain for producing spirogelothoraxone, in which the strong promoter ERG9 is replaced by the weak promoter HXT1 in the strain's genome, and the STO1, AtCPR, tHMG1, and STS genes are integrated. This invention also provides a method for constructing this engineered strain and a method for producing spirogelothoraxone using this engineered strain, which can be used for the preparation of spirogelothoraxone.

The application belongs to the field of new application of functional proteins and mutants thereof, and discloses application of sugarcane ScCBS protein and key domain deletion mutants thereof. The nucleotide sequence of the sugarcane ScCBS protein is shown as SEQ ID NO:1, and the amino acid sequence is shown as SEQ ID NO:2. The key domain deletion mutants of the ScCBS protein are ScCBS△motif1, ScCBS△motif3, ScCBS△motif4 and ScCBS△motif9, which respectively lack 28, 50, 49 and 29 amino acids. The application proves that the ScCBS protein can interact with SCSMV P1 and inhibit the RNA silencing suppressor activity by means of yeast two-hybrid, two-molecular fluorescence complementary experiment and agrobacterium transient expression system; the four mutants all lose the interaction and inhibition activity. Therefore, the ScCBS protein and the key domain deletion mutants thereof can be used for cultivating excellent SCSMV-resistant materials, and have wide application prospects in sugarcane disease-resistant molecular breeding.

The application discloses a low-temperature adaptive carbonic anhydrase mutant HpCA-M3 and application thereof, and relates to the technical field of enzyme engineering. The application screens a low-temperature adaptive carbonic anhydrase mutant HpCA-M3, the amino acid sequence of which is obtained by mutating the serine at the 33rd position of the amino acid sequence of wild-type carbonic anhydrase HpCA into aspartic acid, mutating the serine at the 84th position into glycine, and mutating the proline at the 108th position into serine; the amino acid sequence of the wild-type carbonic anhydrase HpCA is shown as SEQ ID NO. 1. Compared with the wild enzyme HpCA, the carbonic anhydrase mutant HpCA-M3 has higher catalytic activity at low temperature, the esterase activity is increased by 75% at 20 DEG C, and the hydration activity is increased by 23%. The application is helpful to reducing the massive dissolution of CO2 caused by high temperature during industrial carbon sequestration, and can accelerate the enzymatic repair of limestone articles (for example, limestone cultural relics) in high-cold regions, and has important value for promoting carbon capture and limestone cultural relic repair.

The present disclosure describes a recombinant yeast host cell having a first engineered metabolic pathway to convert fructose-6-phosphate (F-6-P) into acetyl-coenzyme A. The first engineered metabolic pathway comprises: a genetic modification for expressing a heterologous phosphoketolase capable of converting fructose-6-phosphate into erythrose-4-phosphate, and at least one genetic modification for decreasing native 6- phosphofructo-2-kinase activity classified under Enzyme Commission No. 2.7.1.105. Optionally, the recombinant yeast host cell comprises a second engineered metabolic pathway for convert acetyl-coA into a fermentation product. The present disclosure further describes a process for making a fermentation product. The process comprises contacting the recombinant yeast host cell described herein with a carbohydrate source under a condition allowing the conversion of at least a part of the carbohydrate into a fermentation product.

The present invention pertains to novel modified terpene synthases and their use for a preparation method for pseudopterosin intermediates and / or pseudopterosins. The method is based on the use of a modified terpene synthase comprising at least one modified amino acid residue, which enables a terpene synthase-catalyzed increased production of pseudopterosin intermediates and / or pseudopterosins from Geranylgeranyl pyrophosphate as starting material. The new modified terpene synthases of this invention allow the production of pseudopterosin intermediates, such as Isoelisabethatriene A, Isoelisabethatriene B, Erogorgiaene, or Seco-Pseudopterosin and / or the production of pseudopterosins, such as Pseudopterosin A, in a cost-efficient, economical, and sustainable manner. Also provided are nucleic acids, encoding for the modified terpene synthases of this invention, as well as expression vectors capable of expressing such nucleic acids and host cells comprising the same.

This invention discloses a walrenene synthase mutant and its applications. Through directed evolution of the enzyme, this invention obtained a walrenene synthase mutant with enhanced enzyme performance. The strain expressing the walrenene synthase mutant achieved a walrenene yield up to 3.41 times that of the wild-type synthase strain. This invention's walrenene synthase mutant improves the production performance of walrenene synthesis in strains, further expanding the application prospects of biosynthetic walrenene.

Provided are a Cinnamomum burmannii monoterpene synthase CbTPS1, an amino acid sequence thereof, a nucleic acid molecule encoding the protein, an use thereof in preparing the monoterpene synthase, and a method for preparing dextrorotatory borneol by using the Cinnamomum burmannii monoterpene synthase CbTPS1.

This invention belongs to the field of enzyme engineering technology, specifically providing a nitrile hydratase mutant and its application. The nitrile hydratase mutant is derived from *Aurantimonas manganoxydans*. Am NHase)SI859A was modified using rational design methods based on its three-dimensional protein structure. The resulting nitrile hydratase mutant ultimately achieved an enzyme activity that was up to 6.74 times higher than that of the wild enzyme, and its stability did not decrease significantly within two hours under heating conditions at 50°C.

The present invention relates to variants of Cas12a nucleases having altered protospacer adjacent motif recognition specificity. The present invention also relates to methods of making CRISPR-CAS nuclease variants and methods of using the variants to modify nucleic acids.

The application discloses a kind of high-yield O-succinyl-L-homoserine recombinant escherichia coli and its construction method and application, belong to genetic engineering and fermentation engineering technical field.The recombinant escherichia coli is expressed by expressing anti-feedback inhibition homoserine O-succinyltransferase metA fbr , aspartate ammonia-lyase aspA, aspartate transaminase aspC, 2 ketoglutarate decarboxylase sucA, dihydrothioctic acid succinyltransferase sucB and efflux protein yjeH, knock out metJ, improve intracellular NADPH availability, to enhance OSH synthesis pathway metabolic flow, precursor succinyl coenzyme A supply and product efflux.Using 5 L bioreactor for fed-batch fermentation, after fermentation 60 h, OSH production is as high as 137.4 g / L, and sugar acid conversion rate reaches 50.5%.The application provides engineering strain and technical realization path for the efficient fermentation of OSH.

This invention discloses a mango terpene synthase gene TPS4 and its applications, the nucleotide sequence of which is shown in SEQ ID NO:1. The amino acid sequence of the protein it encodes is shown in SEQ ID NO:2. The invention also discloses a recombinant expression vector containing the mango terpene synthase gene TPS4, a recombinant engineered strain, and the recombinant mango terpene synthase TPS4; and the application of the mango terpene synthase gene TPS4, its encoded protein, the recombinant expression vector, the recombinant engineered strain, or the recombinant TPS4 in regulating the synthesis of terpenoid compounds from geraniol diphosphate (GPP), neroli diphosphate (NPP), and farnesyl diphosphate (FPP). This invention is the first to clone and verify a terpene synthase gene TPS4 from the Sacred Heart mango genome, capable of simultaneously catalyzing the synthesis of monoterpenes and sesquiterpenes from three substrates: GPP, NPP, and FPP. It has broad application prospects and significant economic value.

This invention belongs to the fields of genetic engineering and enzyme engineering technology, specifically relating to an ornithine decarboxylase mutant, an engineered bacterium, and its application in the production of 1,4-butanediamine. Compared to the protein with the amino acid sequence shown in SEQ ID NO: 1, this mutant contains a mutation at amino acid position 694. The mutant protein exhibits significantly enhanced catalytic activity compared to the parent enzyme and significantly improves the synthesis efficiency of 1,4-butanediamine, providing a new biosynthetic pathway and engineered bacterium for the production of 1,4-butanediamine.

The present application relates to a kind of synthetic hydroxytyrosol multi-enzyme catalytic method.The synthetic hydroxytyrosol multi-enzyme catalytic method described in the present application includes first module reaction, second module reaction, third module reaction in turn;The first module reaction is that glycerol is synthesized pyruvic acid by catalysis, wherein the concentration of the glycerol is 1-5mM;The second module reaction is that pyruvic acid is condensed with catechol and ammonium chloride to be L-levodopa by catalysis;Wherein, the concentration of the catechol is 1-5mM, and the concentration of the catechol is greater than the concentration of the glycerol in the first module reaction;The third module reaction is that L-levodopa is synthesized hydroxytyrosol by catalysis.The synthetic hydroxytyrosol multi-enzyme catalytic method described in the present application, reaction 4 hours hydroxytyrosol molar conversion rate can reach 71.5%, compared with one-pot method has obvious advantage, in the same reaction time molar conversion rate is compared with one-pot method and increased nearly 6 times.

The application provides a saccharomyces cerevisiae engineering bacterium and a preparation method and application thereof. By modifying a saccharomyces cerevisiae strain, the content of santalene and santalol produced by fermentation can be significantly improved.

Disclosed are GLO2 and its use in promoting or inhibiting inflammatory or immune responses. The use of a nucleic acid encoding GLO2, a GLO2 protein, or a promoter of GLO2 in the preparation of a medicament for promoting an inflammatory or immune response, the use of an inhibitor of GLO2 in the preparation of a medicament for inhibiting an inflammatory or immune response, and the use of a detection agent of GLO2 in the preparation of a kit for screening a medicament and / or therapy for an inflammatory or immune response are involved.

This invention discloses a walrenene synthase mutant and its applications. Through directed evolution of the enzyme, this invention obtained a walrenene synthase mutant with enhanced enzyme performance. The strain expressing the walrenene synthase mutant achieved a walrenene yield up to 3.41 times that of the wild-type synthase strain. This invention's walrenene synthase mutant improves the production performance of walrenene synthesis in strains, further expanding the application prospects of biosynthetic walrenene.

The application discloses a kind of biological immobilized enzyme based on yeast surface display technology and the application of synthesis 2-phenylethanol, using yeast surface display technology, for the first time with Saccharomyces cerevisiae surface protein a-lectin as anchor protein, the key enzyme (transaminase, decarboxylase, ethanol dehydrogenase) involved in 2-PE producing Ehrlich pathway in Saccharomyces cerevisiae is separately or jointly carried out biological fixation, enhance substrate channel effect and proximity effect, in vitro one-step catalysis L-phenylalanine synthesis 2-PE, provide yield.The method for fixing multiple enzymes of Ehrlich pathway in the application can be used to reconfigure the biosynthesis pathway of other natural products in vitro, to provide feasible scheme for the rational design of cell factory.

A method and a culture medium for culturing and growing algae utilizing a flue gas and wastewater from a urea plant and / or an ammonia plant are disclosed. In some instances, cultured algae is combined with one or more solid fertilizers to produce bio-fertilizers.

This invention provides a method for preparing bamboo fiber for textiles, belonging to the field of fiber extraction technology. The preparation method includes bamboo strip pretreatment, enzyme immobilization, and bamboo fiber extraction. The bamboo strip pretreatment method is as follows: whole bamboo is taken, crushed and cut, and bamboo strips with a length of 6-8 cm, a width of 0.5-0.7 cm, and a thickness of 0.3-0.5 cm are taken. The strips are ultrasonically treated for 25-35 min, then soaked in NaOH solution for 30-40 min, filtered, washed with deionized water until neutral, and dried to a moisture content of 15-20% to obtain pretreated bamboo strips. The bamboo fiber extracted by this invention has a tensile strength of 1.21-1.28 GPa, a breaking elongation of 4.09-4.17%, an elastic modulus of 37.15-37.26 GPa, and an average enzyme activity retention rate of 66.3-68.4% after 10 reuses.

The application discloses a method for removing chaperone and application thereof. The method for removing chaperone comprises the following steps: after a sample containing a target recombinant protein is loaded on an affinity chromatography column, a washing liquid containing urea and KCl is used to flush the affinity chromatography column, so as to remove the chaperone combined with the target recombinant protein. The method of the application can effectively remove the chaperone of about 60 kDa from an escherichia coli host bacterium by combined washing of urea and KCl, and has the advantages of simple operation, low cost, good chaperone removal effect for various recombinant proteins with different molecular weights, wide universality, and the like, and can significantly improve the purity and quality of the recombinant protein.

This invention relates to a carrier-free immobilized enzyme and its preparation method, wherein the carrier-free immobilized enzyme is a catalytically active inclusion body of the enzyme crosslinked with glutaraldehyde, and the enzyme's end (especially the N-terminus) is tagged with histidine-leucine-histidine-leucine-histidine-leucine.

The application discloses a new method for synthesizing N-methyl-beta-alanine by catalysis of aspartate ammonia-lyase, and comprises the following steps: taking acrylic acid and methylamine as substrates, and using aspartate ammonia-lyase or a mutant thereof with an amino acid sequence shown in SEQ ID NO:1 to catalyze a synthesis reaction to obtain N-methyl-beta-alanine. The application provides a biological catalysis path for the production of N-methyl-beta-alanine.

This application discloses a fatty acid photodecarboxylase mutant, its preparation method, and its applications, relating to the field of protein engineering technology. The amino acid sequence of the fatty acid photodecarboxylase mutant of this application comprises a sequence obtained by mutating the 624th amino acid residue site in the sequence shown in SEQ ID NO.1, wherein the mutation type at the 624th amino acid residue site is selected from any one of A624C, A624N, A624Q, and A624S. Compared with the wild type, the preferred mutant of this application exhibits 2-fold and 1.3-fold increased enzyme activity and blue light tolerance, respectively. It achieves a conversion rate of over 90% for the long-chain fatty acid palmitic acid within 8 hours at 30°C, significantly enhancing the enzyme's stability and catalytic efficiency under blue light irradiation. This helps overcome the bottleneck of photoinactivation caused by blue light in the industrial application of fatty acid photodecarboxylase, and has excellent application prospects in biofuel synthesis and green chemistry.

The present application belongs to the technical field of biological medicine, and relates to a polypeptide targeting the interaction interface between PGAM5 and HADHA protein and application thereof. The polypeptide is derived from a key binding region of HADHA, can specifically bind to PGAM5, thereby competitively inhibiting the interaction between PGAM5 and HADHA, and relieving the inhibitory effect of PGAM5 on the activity of HADHA. The polypeptide can regulate the HADHA / beta-HB / HDAC / IL-10 signal pathway, up-regulate the expression of anti-inflammatory factors and inhibit the expression of pro-inflammatory factors, thereby playing an anti-inflammatory role. The polypeptide is applied to the preparation of a drug for preventing and / or treating inflammatory diseases, especially has application value in the prevention and treatment of inflammatory and immune-related diseases such as acute lung injury caused by sepsis, and can be used for preparing a related therapeutic drug.