65 results about "Archaeal Viruses" patented technology

The invention belongs to the technical field of biosynthesis, and particularly relates to a strain for producing sclareol as well as a construction method and application of the strain. In order to solve the problem of microbial synthesis of sclareol, phosphoketolase in an NOG pathway is mutated and optimized, phosphoketolase NaXpk from nanoarchaea archaeon is subjected to site-directed mutagenesis, a sclareol synthesis pathway is constructed in an escherichia coli host, the influence of NaXpk before and after mutation on the yield of sclareol is compared, and the yield of sclareol is improved. The S472F mutant and the S472Y mutant are determined, the S472F mutant and the S472Y mutant are applied to a sclareol production path, the sclareol yield of the finally obtained production strains can reach 623.3 mg / L and 655.2 mg / L, and the effect is remarkable.

The invention provides a microbial composite remediation liquid organic fertilizer and a preparation method thereof, and aims to solve the problems of single function, limited pollution abatement capability and the like of a traditional liquid fertilizer. Through synergism of strains of bacillus velezensis, bacillus methylotrophicus, pseudomonas putida and methanogenic archaea, recycling of agricultural wastes such as straw and mushroom dreg compost and decomposition, extraction of humic acid through enzymolysis and a formula of green additives (mineral source potassium fulvate, sophorolipid and the like), chemical passivation of heavy metals, pesticide degradation and soil improvement are realized; and dynamic fermentation control and membrane separation processes are combined, so that the method has the characteristics of efficient repair and low-carbon sustainability, and an integrated solution is provided for pollution treatment of complex farmlands.

Provided herein are modified Archaeal family B polymerases derived from the Archaeal microorganism Pyrococcus abyssi that exhibit improved incorporation of nucleotide analogues utilized in DNA sequencing.

The application discloses a kind of folded plate type anaerobic biological reactors, including box, the water inlet and water outlet are equipped in two opposite side walls of the box;Box is sequentially provided with at least two reaction compartments, the top of upstream reaction compartment is communicated with the lower part of downstream reaction compartment, the water inlet is communicated with the lower part of first reaction compartment of head end, the water outlet is communicated with the top of last reaction compartment of terminal end;The side wall of the box above the water outlet is further provided with exhaust port;Each reaction compartment is respectively provided with biological filler module, and the biological filler module includes polyurethane foam filler.The application further discloses an application of the folded plate type anaerobic biological reactor in treating aquaculture wastewater.The technical scheme of the application has the remarkable effects of significantly improving oxygen resistance, outstanding archaea enrichment effect, double improvement of treatment efficiency and biogas quality, and significant resource value.

This invention discloses methanogenic lyases with the same protein family annotation as PeiR lyases and their applications. These enzymes are obtained through homology screening based on the known characteristics of PeiR lyases targeting peptide bonds in the methanogenic cell wall, using the Pfam protein family database annotation. PeiR lyase proteins participate in the biological process of hydrolyzing archaea cell walls, effectively killing methanogens and reducing methane production. Proteins homologous to this protein have potential methane-reducing potential. This invention integrates rumen microbial metagenomic sequencing data, uses a series of bioinformatics software to screen methanogenic viral proteins, and combines Pfam functional domain annotation for homology analysis, ultimately identifying a series of lyases. These lyases were successfully expressed in a prokaryotic expression system, and in vitro gas production experiments also showed that the crude enzyme solution significantly reduced methane production.

The invention discloses a specific amplification primer pair for human intestinal archaea flora 16S rRNA amplicon sequencing and application of the specific amplification primer pair, and belongs to the technical field of biology.The primer pair is designed for 16S rRNA high-throughput sequencing and can specifically amplify human intestinal archaea, the length of an amplification product is about 385bp, and the amplification product covers a V4-V5 variable region of a 16S rRNA gene; the invention also optimizes amplification conditions of the primer pair, and experimental results show that the primer pair is superior to a conventional universal primer for bacteria and archaea in the aspect of amplifying species diversity and species abundance of the intestinal archaea, and a standardized method is provided for efficient and accurate amplification of intestinal archaea florae.

The invention relates to the field of biological fermentation, in particular to a method for efficiently preparing anti-aging ergothioneine through fermentation of compound bacteria combined with bolete and application thereof.The method comprises the steps that halophilic bacillus SC-1, halophilic archaea IR-2 and thermophilic casein dissolving bacteria PC-3 are mixed according to the volume ratio of 1: 1: 1, bolete and pleurotus citrinopileatus are used as raw materials, and a mixture is prepared through fermentation; fermenting for 120 hours in an optimized basic culture medium under the conditions that the temperature is 30 + / -0.5 DEG C, the speed is 150rpm + / -10rpm and the pH value is 5.8 + / -0.2; through a step-by-step process of purifying EGT by using D101 resin and purifying RNET-1 by using DEAE-52 resin, a double-active-component extract is synchronously obtained. Through the synergistic effect of three strains with complementary functions, an efficient system of raw material degradation, carbon source supply, double precursor synthesis and double active component accumulation is constructed, synchronous high yield of ergothioneine and novel antioxidant protein RNET-1 is realized for the first time, and the blank of single active component in the prior art is filled.

This invention discloses a methanogenic lyase homologous to PeiR lyase and its applications. The PeiR lyase protein, targeting the peptide bonds of the methanogenic cell wall, participates in the biological process of hydrolyzing the archaea cell wall, effectively killing methanogens and reducing methane production. Proteins homologous to this lyase possess potential methane-reducing potential. This invention utilizes a series of bioinformatics software to identify protein sequences encoded by methanogenic viruses from a large amount of rumen microbial virome sequencing data, and analyzes the structural homology between these proteins and PeiR. Ultimately, this invention successfully obtained a series of methanogenic lyases homologous to PeiR. To achieve the expression of these lyases, this invention designed multiple primers to synthesize the target sequences and successfully expressed them in a prokaryotic expression system. In vitro gas production experiments with the crude enzyme solution showed that these lyases are effective in reducing methane production.

The invention relates to the technical field of soil conditioners and applications thereof, and discloses a soil carbon sequestration-pollution reduction-yield increase multi-effect synergistic microbial inoculum and an application thereof in ploughing ability improvement, the soil carbon sequestration-pollution reduction-yield increase multi-effect synergistic microbial inoculum comprises an organic pollutant degradation functional flora, a soil carbon sequestration functional flora and a small molecular organic matter production functional flora; the organic pollutant degrading functional flora comprises efficient pollutant degrading bacteria; the soil carbon sequestration functional flora comprises ammonia oxidizing bacteria, ammonia oxidizing archaea, complete ammonia oxidizing bacteria, sulfate reducing bacteria, iron reducing bacteria, rhizobium and photosynthetic flora; the micromolecular organic matter production functional flora comprises heterotrophic nitrification-aerobic denitrification bacteria, zoogloea, phosphorus-accumulating bacteria and photosynthetic bacteria. According to the scheme, multiple florae are specifically arranged in the synergistic agent, and through the synergistic interaction effect of the florae, the effects of the synergistic agent in multiple fields of soil carbon sequestration, pollution reduction and yield increase are effectively improved.

This invention discloses an anti-phage system based on ESCRT-III and its applications, relating to the field of biotechnology. This invention utilizes the coding genes for two structural proteins of the Hoda_Snf7_1 and Hoda_Snf7_2 from the Hoda archaea ESCRT-III. Hoda_ snf7_1 and Hoda_snf7_2 This invention is applied to antiphage systems. Compared with existing technologies, the main advantages of the antiphage system of this invention are: (1) it consists of only two genes, making system construction simple and quick; (2) it is composed of structural proteins, making it less prone to losing antiphage activity due to gene mutations; (3) it has a broad-spectrum antiphage effect; and (4) it does not affect the normal growth of host bacteria. This system expands the understanding of antiphage systems and provides new ideas and technical means for preventing and controlling phage contamination in industrial microbial production processes.

The present application relates to a kind of plasmid construction method for identifying and deleting large fragment non-essential gene region and its application in gene editing.The plasmid is the gene editing plasmid pMHAGE containing mini-CRISPR cluster and multiple donor DNA fragments.The plasmid carries artificial CRISPR cluster and multiple donor DNA simultaneously, and multiple donor DNA can carry out multiple forms of gene editing to prokaryote using CRISPR-Cas system through homologous recombination, the identification and deletion of large fragment non-essential gene region, realize the simplification of prokaryote genome.The plasmid application host range is wide, and all bacteria and archaea using CRISPR-Cas system can be operated;It can be used to identify large fragment non-essential gene region and editable non-essential gene region combination on genome;With the ability of screening growth phenotype and the optimal non-essential gene region large fragment deletion of genome simplification chassis cell resistance;Simple process, short time cycle, greatly reduce the workload when prokaryote genome simplification.

The application discloses an application of a crenarchaeota-derived Argonaute protein or a coding gene thereof in RNA interference. The Argonaute protein is named as HrAgo1, and the amino acid sequence is shown as SEQ ID No. 1. The application finds a new crenarchaeota-derived Argonaute protein, which is named as HrAgo1. Different from known prokaryote Argonaute proteins, the HrAgo1 has two characteristics. First, the HrAgo1 can use short-chain RNA as a guide chain to cut mRNA paired with the sequence. Therefore, a gene interference method based on the HrAgo1 is obviously different from other applications based on prokaryote Argonaute proteins. Second, the HrAgo1 can exert the function at 9-71.1 DEG C, and can be applied to cells growing at different temperatures.

The application discloses a non-conventional methanogenic archaea and belongs to the technical field of microorganisms. The methanogenic archaea (Methanoproducendum sp.) DRTY01 has a preservation number of CCTCC NO: M 2025822 and is preserved in the China Center for Type Culture Collection, is the first archaea with a methanogenic function isolated at present, and is a new genus representative in the Archaeoglobaceae family. The methanogenic archaea DRTY01 is also the only strain capable of efficiently utilizing methanol to produce methane under the condition of 85 DEG C ultra-high temperature, thereby providing an efficient, environment-friendly and feasible technical scheme for energy supply under specific scenes such as an ultra-high temperature fermentation tank.

The invention provides a cellobiose phosphorylase mutant capable of synthesizing laminobiose, which is characterized in that substrate specificity of cellobiose phosphorylase from thermophilic archaea is changed, wild cellobiose phosphorylase with an amino acid sequence as shown in SEQ ID NO: 2 is used as starting protein, multiple rounds of site-directed mutagenesis are carried out on the starting protein through rational design, and the cellobiose phosphorylase mutant capable of synthesizing laminobiose is obtained. The mutant and a wild type CBP amino acid sequence SEQ ID NO: 2 have at least 90% identity, the mutant has the activity of catalyzing glucose and glucose 1-phosphoric acid to synthesize laminobiose, the mutant is super-heat-resistant laminobiose phosphorylase, the dominant mutant is applied to an in-vitro multi-enzyme system for catalytically producing laminobiose by taking starch as a substrate, and the yield of the laminobiose is improved. And the laminobiose can be synthesized at high temperature, high conversion rate and high efficiency.

The invention relates to the field of wine brewing, in particular to a microbial symbiont-based wine brewing method, which comprises the following steps: preparing a functional symbiont containing specific acid-resistant archaea and high-yield enzyme yeast, and layering and inoculating the functional symbiont into a pretreated raw material mixture according to a predetermined inoculum size, in the fermentation process, the temperature, the ventilation capacity and the pH value are dynamically regulated and controlled according to detection indexes, and finally, an intelligent enzymolysis-ultrasonic coupling aging process is carried out on the base liquor, and specific steps and parameters of symbiont preparation, raw material pretreatment, aging processes and the like are involved. The technical effects of effectively inhibiting the lactic acid bacteria, improving the saccharifying enzyme activity, accurately regulating and controlling the fermentation process, improving the flavor and quality of the wine, accelerating the aging process of the wine and the like are achieved.

Provided herein are archaeal B -family polymerase variants with improved replication fidelity compared to wild-type archaeal B-family polymerases and methods of use.

This invention discloses a reactor, method, and application for the graded enrichment of functional microorganisms, belonging to the field of wastewater biological treatment technology. The method combines a composite packing frame, a two-stage sedimentation tank, and controlled operating conditions to create a multi-gradient distribution environment of sludge settling properties, sludge age, dissolved oxygen, and substrate concentration under continuous flow operation. This forms multiple micro-niche areas, such as suspended zones, packing attachment zones, and sedimentation zones, thereby achieving graded enrichment of different functional microorganisms within a single reactor. Each zone is enriched with functional microbial communities including organic matter hydrolyzing bacteria, ammonia-oxidizing archaea, aerobic denitrifying bacteria, autotrophic denitrifying bacteria, iron-oxidizing-reducing bacteria, and sulfur-oxidizing-reducing bacteria, achieving synergistic coupling of multiple metabolic pathways. This invention achieves multifunctional microbial synergy without the need for multi-stage reactors or complex process segments, improving pollutant removal efficiency and multi-element cycling processes. It has the advantages of stable operation, strong adaptability, and ease of engineering application.

The invention relates to an archaea amoA gene absolute quantitative detection method, primers and a kit, and by using the primers with the sequences of amoAF: GCAGGCGACTATCTCTAC and amoAR: CATATACGTTGCCGTTCCT, the advantages of single amplification product, reasonable CT value interval and large archaea amoA gene amplification quantity are achieved.

This invention belongs to the field of biosynthesis technology, specifically relating to a strain for producing perillaldehyde, its construction method, and its application. Addressing the microbial synthesis problem of perillaldehyde, this invention mutates and optimizes the phosphotransketase in the NOG pathway, using a strain derived from nanoarchaemonas (…). Nanoarchaeota archaeon Site-directed mutagenesis was performed on the phosphoketolase NaXpk, and a perillaldehyde synthesis pathway was constructed in E. coli. The effects of NaXpk mutation on perillaldehyde yield were compared to identify the S472F and S472Y mutants. These mutants were then applied to the perillaldehyde production pathway, and the resulting production strains achieved perillaldehyde yields of 623.3 mg / L and 655.2 mg / L, respectively, demonstrating significant effectiveness.

To provide RNA-guided endonuclease polypeptides referred to as CasX polypeptides (also referred to as CasX proteins).SOLUTION: The present disclosure provides CasX proteins, nucleic acids encoding the CasX proteins, and modified host cells comprising the CasX proteins and / or nucleic acids encoding the same. CasX proteins are useful in a variety of applications provided. The present disclosure further provides CasX guide RNAs that bind to and provide sequence specificity to the CasX proteins, nucleic acids encoding the CasX guide RNAs, and modified host cells comprising the CasX guide RNAs and / or nucleic acids encoding the same. CasX guide RNAs are useful in a variety of applications provided. The present disclosure also provides archaeal Cas9 polypeptides and nucleic acids encoding the same, as well as their associated archaeal Cas9 guide RNAs and nucleic acids encoding the same.SELECTED DRAWING: Figure 30-4

The invention belongs to the technical field of forensic medicine and biological medicine, discloses a method for forensic medicine identification of corpses in water based on metagenomics markers and application, and aims to solve the problems that existing drowning diagnosis is insufficient in accuracy and the deduction of post-death submerging time (PMSI) is limited. The method comprises the following steps: collecting a corpse lung tissue in water and a water sample in a corresponding water area, and obtaining a microbial DNA sequence by adopting a metagenome sequencing technology; the method comprises the following steps: processing data through bioinformatics analysis (such as fastp quality control, host sequence removal by KneadData and Kraken2 species annotation), and selecting and screening species level microbial markers in combination with a random forest algorithm and Boruta characteristics; and respectively establishing a drowning diagnosis model and a PMSI inference model based on the screened markers, and verifying the performance of the models through indexes such as AUC and MAE. Complete microbial communities of bacteria, eukaryotes (including fungi), archaea and viruses in lung tissues of corpses in water are comprehensively analyzed for the first time, 17 bacterial markers and 9 eukaryote markers are screened out for drowning diagnosis (verification experiment bacterial model AUC = 1, the accuracy rate is 89.29%, eukaryote model AUC = 0.95, the accuracy rate is 87.5%), and 17 markers are screened out for PMSI inference (integrated model MAE = 0.66 days). According to the method, the species resolution reaches the species level, the method is not influenced by amplification bias, the method is suitable for different water area environments, and an efficient and objective forensic medicine tool is provided for identifying the cause of the dead body in water and deducing the submerged time after the dead body.

This invention belongs to the field of genetic engineering and relates to a fusion-type halophilic archaea chitinase, its preparation method, and its applications. Based on chitinase domains from different halophilic archaea, this invention uses overlap extension PCR technology to fuse a chitin-binding domain with strong substrate-binding ability with a catalytic domain with high catalytic activity, thereby constructing a fusion gene. chiB A fusion-type chitinase was obtained through expression and purification. This fusion-type chitinase possesses both catalytic activity and substrate-binding ability, exhibiting excellent degradation efficiency for natural chitin substrates, as well as good salt tolerance and resistance to metal ions, organic solvents, and surfactants. The prepared enzyme can not only be used for chitin degradation, but the degradation products can also be used to prepare chitosan oligosaccharides or N-acetylglucosamine, showing broad market application prospects.

The present invention relates to a method and apparatus for controlling the feed rate of at least one reactant in a methanation process. In particular, the present invention relates to a method and apparatus for iteratively controlling the feed rate of at least one reactant in a methanation process carried out in a reactor, such as an industrial-sized reactor and / or a reactor assembly according to the present invention. More specifically, the present invention refers to a method and apparatus for iteratively controlling the feed rate of at least one reactant in a methanation process in an aqueous culture medium containing a biocatalyst such as an archaea.

A system and method for fixing carbon dioxide by fermentation are provided. More specifically, the disclosure includes the steps of producing methane by fermenting carbon dioxide with methanogenic archaea and producing a single-cell protein supplement. The disclosure further provides integration of methane-producing fermentation with additional methods to achieve improved carbon efficiency.

Systems and methods for immobilizing carbon dioxide by fermentation are provided. More specifically, the present disclosure includes fermenting carbon dioxide to methane by methanogenic archaea and producing a single cell protein nutritional supplement. The present disclosure also provides integration of methanogenic fermentation with other methods to achieve improved carbon efficiency.

This invention belongs to the field of genetic engineering and relates to a method for preparing amylase derived from halophilic archaea and its application; based on halophilic archaea... Haloarcula sp. ZJK10-12、 Haladaptatus The genes for α-amylase, maltose amylase, and glucoamylase were obtained from the whole genome sequence of sp. ZJK29-3. Recombinant plasmids were constructed using primer design and molecular cloning, and transformed into prokaryotic host bacteria to obtain the corresponding recombinant strains. The bacteria were enriched and cultured in Hv-YPC liquid medium, and the cells were collected and lysed by sonication. α-amylase, maltose amylase, and glucoamylase were obtained using nickel column affinity chromatography. The amylases exhibit excellent enzymatic properties and outstanding salt tolerance. Their application in the fermentation of high-salt, dilute soy sauce can improve raw material utilization and product quality, demonstrating good industrial applicability and promotional value.

The invention provides an intestinal flora composition for preventing or treating osteoporosis and application thereof, and relates to the technical field of biology. According to the invention, causal association analysis is carried out on whole genome heritable variation data and intestinal flora data of East Asian population through a Mendel randomization method, and the intestinal flora for preventing or treating osteoporosis is screened out; comprising acid amide bacteria enterobacter, bacillus haulis, kiwifruit bacteria, archaea extensa, streptococcus ricini and lysobacter pitermannii, and an accurate microbial intervention target is provided for prevention and intervention of osteoporosis. The intervention mode based on microbiomics has the advantages of noninvasiveness, high safety, good tolerance and the like, can be used for a long time, has no drug accumulation risk, and is especially suitable for early prevention and chronic management scenes of osteoporosis.

The invention discloses a difunctional expansin protein mutant as well as a coding gene and application thereof, and belongs to the technical field of biological genetic engineering. According to the invention, a protein PFLoos1 with bifunctional activity is obtained from a metagenome of rumen microorganisms, and mutants PFLoos1-S1, PFLoos1-S2 and PFLoos1-S3 are obtained by carrying out site-specific mutagenesis on key amino acids in a substrate binding pocket or a surface hydrophobic region of the protein PFLoos1. The mutant provided by the invention has the function of inhibiting the metabolic activity of rumen methanogenic archaea on the basis of keeping the lignocellulose degradation promoting capability. The mutant provided by the invention not only can be applied to preparation of an enzyme preparation for promoting cellulose degradation, but also can be applied to a feed additive for inhibiting ruminant rumen methane generation, and has important practical value for improving feed energy utilization efficiency and low-carbon breeding of animal husbandry.

The present invention describes a nanoarchaeosomal composition and method for delivering biomolecules comprising nanoparticulate vesicles of archaeal lipids, wherein the biomolecule is encapsulated in the vesicles formed by archaeal lipids. Further, the therapeutic drug encapsulated can be a breast cancer targeting drug, quercetin. The nanoarchaeosomal composition is colloidal in nature, stable at high temperatures, and under oxidative stress conditions. Further, exhibits higher loading efficiency and bioavailability. It also exhibits lesser cytotoxicity.