115 results about "Archaeal Viruses" patented technology

The invention discloses methanogen lyase homologous with a PeiR lyase sequence and application of the methanogen lyase, and aims to solve the problem that PeiR lyase protein of a methanogen cell wall peptide bond can participate in a biological process of hydrolyzing archaea cell walls, effectively kill methanogen and reduce methane production, and protein homologous with the PeiR lyase protein has potential methane reduction potential. According to the invention, a series of biological information software is utilized to identify protein sequences coded by methanogens virus from rumen microbiome sequencing data, and sequence homology between the proteins and PeiR protein is further analyzed to obtain a series of methanogens lyases. In order to realize the expression of the lyase, multiple segments of primers are designed to synthesize a target sequence, and the target sequence is successfully expressed in a prokaryotic expression system. Through the detection of crude enzyme liquid, the effectiveness of the lyase in the aspect of reducing the methane yield is verified in an in-vitro gas production test.

The invention discloses methanogen lyase homologous with high-dimensional characteristics of PeiW lyase and application of the methanogen lyase, and aims to solve the problem that PeiW lyase protein of methanogen cell wall peptide bonds can participate in the biological process of hydrolyzing archaea cell walls, effectively kill methanogen and reduce methane production, and protein homologous with the protein has potential methane reduction potential. According to the method, a series of bioinformatics software is utilized, a protein sequence coded by methanogens virus is identified from a large amount of rumen microorganism virus group sequencing data, the high-dimensional homology of a deep learning model between the protein sequence and PeiW is analyzed, and two methanogens lyases similar to the high-dimensional characteristics obtained by the PeiW model are obtained. In order to realize the expression of the lyase, multiple segments of primers are designed to synthesize a target sequence, and the target sequence is successfully expressed in a prokaryotic expression system. Through detection of a crude enzyme solution, the effectiveness of the lyase in the aspect of reducing the yield of methane is verified in an in-vitro gas production test.

The invention discloses methanogen lyase with the same protein family annotation as PeiR lyase and application of the methanogen lyase, and the methanogen lyase is obtained through homologous screening of Pfam protein family database annotation based on known PeiR lyase characteristics aiming at methanogen cell wall peptide bonds. The PeiR lyase protein can participate in the biological process of hydrolyzing archaea cell walls, methanogens are effectively killed, methane production is reduced, and the protein homologous with the protein has potential methane reduction potential. According to the invention, by integrating metagenome sequencing data of rumen microorganisms, screening methanogenic bacterium virus protein by using series bioinformatics software, and combining Pfam functional domain annotation to carry out homologous relationship analysis, a series of lyase is finally identified and is successfully expressed in a prokaryotic expression system, and meanwhile, an in-vitro gas production experiment result also shows that the methanogenic bacterium virus protein can be successfully expressed in a prokaryotic expression system. The crude enzyme liquid can significantly reduce the methane generation amount.

The invention discloses methanogen lyase homologous with high-dimensional characteristics of PeiR lyase and application of the methanogen lyase and aims to solve the problems that PeiR lyase protein of methanogen cell wall peptide bonds can participate in the biological process of hydrolyzing archaea cell walls, methanogen is effectively killed, methane production is reduced, and protein homologous with the protein has potential methane reduction potential. According to the invention, a series of biological information software is utilized to identify protein sequences coded by methanogen viruses from a large amount of rumen microbiome sequencing data, and high-dimensional homology between the proteins and PeiR proteins based on a deep learning model TM-Vec is further analyzed, so that a series of methanogen lyases are obtained. In order to realize the expression of the lyase, multiple segments of primers are designed to synthesize a target sequence, and the target sequence is successfully expressed in a prokaryotic expression system. Through the detection of crude enzyme liquid, the effectiveness of the lyase in the aspect of reducing the methane yield is verified in an in-vitro gas production test.

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 discloses an application of ribonuclease SiRe0902 with high efficiency and thermal stability in specific recognition of a GG site. The amino acid sequence of the gene is as shown in SEQ ID NO.2, and the nucleotide sequence of the gene is as shown in SEQ ID NO.1. The ribonuclease SiRe0902 has very efficient and specific RNA (Ribonucleic Acid) hydrolysis activity, and does not have non-specific cutting on DNA (Deoxyribose Nucleic Acid) at all. In addition, the enzyme is derived from thermophilic archaea, is detected to have good thermal stability, and has very good RNA cleavage activity under the conditions of 37 DEG C and 65 DEG C. Through detection, the enzyme can specifically recognize a GG sequence for cutting, and also has high cutting activity for the GG sequence in a secondary structural region. Therefore, the ribonuclease SiRe0902 is novel ribonuclease with thermal stability, has extremely high RNA hydrolytic activity under normal-temperature and high-temperature conditions, and meets the requirements in the fields of clinic, production, molecular cloning and the like.

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 invention discloses an amylase mutant. The amino acid sequence of the amylase mutant is shown as SEQ ID NO: 2 in a sequence table. The mutant is obtained by carrying out transformation and screening of protein engineering on the high-temperature alpha-amylase from thermococcus eurythermalis, and the high-temperature alpha-amylase is obtained by carrying out transformation and screening of protein engineering on the high-temperature alpha-amylase from thermococcus eurythermalis.

The present invention provides a method for producing an amino acid by fermentation in a bioreactor wherein the bioreactor comprises a methanogenic microorganism in a fermentation broth, the method comprising at least the steps of: producing an amino acid from the methanogenic microorganism under conditions such that the methanogenic microorganism produces the amino acid; the bioreactor is supplied with a gaseous carbon source comprising carbon dioxide and / or carbon monoxide, a nitrogen source, and preferably a sulfur source, wherein the amino acid comprises normal valine. Preferred methanogens are, for example, archaea selected from the group consisting of Methanothermophiles, Methanothermophiles, and Methanococci. Also provided are fermentation broths comprising a methanogenic microorganism and normal valine, as well as bioreactors comprising the fermentation broths.

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 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.

The invention belongs to the technical field of kitchen waste recycling, and particularly relates to polyaniline magnetic biochar prepared on the basis of biogas residues and application of the polyaniline magnetic biochar in promoting enrichment of anaerobic co-digestion synergistic functional bacteria and methanogenic archaea in an anaerobic co-digestion system. According to the method, a proper amount of phosphate is added into the dry anaerobic digestion system of the kitchen garbage to activate the magnetic enteromorpha hydrothermal carbon so as to optimize the microbial community structure of the digestion system, enhance the microbial activity and enrich functional microorganisms such as clostridium and methane bag-shaped bacteria, so that the dry anaerobic digestion process of the kitchen garbage is effectively promoted, and the digestion efficiency of the kitchen garbage is improved. And the biogas yield is improved. And the method has great significance for realizing reduction, harmlessness and recycling of the kitchen garbage.

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.

The invention discloses methanogenic archaea, and belongs to the technical field of microorganisms. The preservation number of the methanogenic archaea DRTY01 is CCTCC NO: M 2025822, the methanogenic archaea DRTY01 is preserved in the China Center for Type Culture Collection, and the methanogenic archaea DRTY01 is a currently separated first strain of archaea with a methanogenic function, and is a new representative in the archaea family. The methanogenic archaea DRTY01 is also the only strain capable of efficiently utilizing methanol to produce methane at the ultra-high temperature of 85 DEG C at present, so that an efficient, environment-friendly and feasible technical scheme is provided for energy supply in specific scenes such as an ultra-high temperature fermentation tank.

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 invention discloses methanogen lyase homologous with high-dimensional characteristics of PeiP lyase and application of the methanogen lyase, and aims to solve the problem that PeiP lyase protein of methanogen cell wall peptide bonds can participate in the biological process of hydrolyzing archaea cell walls, effectively kill methanogen and reduce methane production, and protein homologous with the protein has potential methane reduction potential. According to the method, a series of bioinformatics software is utilized, a protein sequence coded by methanogens virus is identified from a large amount of rumen microorganism virus group sequencing data, the high-dimensional homology of a deep learning model TM-Vec between the protein sequence and PeiP is analyzed, and the methanogens lyase homologous with the PeiP on the high-dimensional feature level of the model is obtained. According to the invention, multiple segments of primers are designed to synthesize a target sequence, the expression of the lyase is successfully realized in a prokaryotic expression system, and the effectiveness of the novel lyase in the aspect of reducing the yield of methane is verified in an in-vitro gas production test by detecting a crude enzyme solution.

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 invention relates to the technical field of microorganisms, and in particular relates to a prevotella bruceli B14 capable of improving the milk yield and the milk-fat percentage of ruminants and application of the prevotella bruceli B14. The invention discloses a strain of prevotella bruceli B14, and the Latin name of the prevotella bruceli B14 is prevotella bruceli B14. The preservation number of the strain is CCTCC NO: M 20242730. The prevotella bruceli B14 provided by the invention can increase the milk yield and the milk fat percentage of ruminants by increasing the concentration of total volatile fatty acid, acetic acid, propionic acid and valeric acid in the rumen of the ruminants. In addition, the compound also has the function of synthesizing nicotinic acid, and can activate an mTORC1 signal channel in the mammary gland of the ruminant in a GPR109A-dependent manner and promote the expression of lipid synthesis genes so as to promote milk fat synthesis. In addition, the prevotella buchneri B14 also has the function of reducing the abundance of methanogenic archaea in rumen, so that the prevotella buchneri B14 is expected to improve the energy utilization efficiency by reducing the methane emission of ruminants.

The invention provides an RPA-PfAgo detection system for detecting beef and application of the RPA-PfAgo detection system, and belongs to the technical field of food detection. The RPA-PfAgo detection system disclosed by the invention comprises an RPA specific primer pair, gDNA1-1 and gDNA1-2 sequences of PfAgo protein and a probe, wherein the RPA specific primer pair is designed aiming at a beef gene; and the gDNA1-1 and gDNA1-2 sequences of the PfAgo protein. According to the invention, the high efficiency and specificity of RPA are utilized, and Argonaute protein from extreme thermophilic archaea is combined, so that the detection capability is enhanced. And the RPA-PfAgo detection system is optimized, so that the specificity and the sensitivity of beef recognition are enhanced. By detecting meat products and artificially adulterated meat samples in a supermarket, the universal applicability of the system is tested. A practical solution is provided for rapid on-site detection, and verification of accurate identification of beef in commercial and supervision environments is enhanced.

The invention provides an RPA-PfAgo detection system for detecting chicken and application of the RPA-PfAgo detection system, and belongs to the technical field of food detection. The RPA-PfAgo detection system disclosed by the invention comprises an RPA specific primer pair, gDNA45-1 and gDNA45-2 sequences of a PfAgo protein and a probe, wherein the RPA specific primer pair is designed aiming at a chicken gene; and the gDNA45-1 and gDNA45-2 sequences of the PfAgo protein. According to the invention, the high efficiency and specificity of RPA are utilized, and Argonaute protein from extreme thermophilic archaea is combined, so that the detection capability is enhanced. And by optimizing an RPA-PfAgo detection system, the specificity and the sensitivity of chicken recognition are enhanced. By detecting meat products and artificially adulterated meat samples in a supermarket, the universal applicability of the system is tested. A practical solution is provided for rapid on-site detection, and verification of accurate recognition of chicken in commercial and supervision environments is enhanced.

The invention discloses a halophilic genetically engineered bacterium for synthesizing beta-carotene as well as a construction method and application of the halophilic genetically engineered bacterium. The halophilic genetically engineered bacterium H.volcanii is constructed by cloning a lycopene beta-cyclase gene Hma-LCYb coded by a halophilic archaea marisportui genome to an expression vector pTA1228 and then transforming the lycopene beta-cyclase gene Hma-LCYb into a halophilic archaea Haloferx volcanii cell. The engineering bacterium can be used for synthesizing beta-carotene, and the yield of the engineering bacterium is up to 0.26 mg / g of dry cell weight. The halophilic genetically engineered bacterium H.volcanii disclosed by the invention solves the problems of strain function deficiency, insufficient yield and safety in the production of natural beta-carotene, has high efficiency, economical efficiency and environmental protection property, and has remarkable industrial application value.

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 discloses methanogen lyase homologous with a PeiP lyase structure and application of the methanogen lyase, and the methanogen lyase is screened from rumen microbiome sequencing data through a structure comparison method based on known PeiP lyase protein characteristics. The PeiP lyase protein can participate in the biological process of hydrolyzing archaea cell walls, methanogens are effectively killed, methane production is reduced, and the protein homologous with the protein has potential methane reduction potential. Protein sequences coded by methanogens viruses are extracted from rumen microbiome sequencing data, the structural similarity between the proteins and PeiP proteins is analyzed by using a bioinformatics tool, and finally two lyases are successfully identified. In order to realize the expression, multiple segments of primers are designed and synthesized to construct a target sequence, and high-efficiency expression is realized in a prokaryotic expression system. In-vitro gas production experiments show that the crude enzyme liquid can significantly reduce methane production.