61 results about "Organomercurial lyase" patented technology

Methods for the microbial production of para-hydroxycinnamic acid (pHCA) and cinnamic acid (CA) are provided. Microbes producing either tyrosine or phenylalanine are grown in the presence of either tyrosine ammonium lyase or phenylalanine ammonium lyase respectively where some part of the fermentation is accomplished at alkaline pH. The process results in greater yields and higher rates of para-hydroxycinnamic acid (pHCA) and cinnamic acid (CA) production as compared with fermentation exclusively at physiological pH.

The invention provided a fore treatment process of the enzyme rolling pile for the fabric which belongs to the fore treatment process of the dyeing and finishing field. The process is: first to impregnate the fabric into the mixture of the alkali pectinase liquid, neutral proteinase liquid and xylanase liquid, then to keep the temperature, next to wash with the hot water (killing the enzyme) and the cool water last to blanch with the H2O2. The pectinase is produced by the Bacillus subtilis WSHB04-03. The process has many merits such as high efficiency, low work output and the energy conservation and so on.

The present invention is directed to novel 2-amino-3,4-dihydro-pyrido[3,4-d]pyrimidine derivatives, pharmaceutical compositions containing them and their use in the treatment of Alzheimer's disease (AD) and related disorders. The compounds of the invention are inhibitors of β-secretase, also known as β-site cleaving enzyme and BACE, BACE1, Asp2 and memapsin2.

The invention discloses a recombinant alginate lyase and a construction method and an application thereof, which belong to the technical field of biology. The alginate lyase Aly-Cob is derived from anenvironment sample (a mixture of sea-tangle stacking place and factory production waste material). By using a gene engineering technology, a gene of the alginate lyase is cloned to an expression vector, and a recombinant bacterial strain for heterogenous expression of the alginate lyase is obtained. The alginate lyase Aly-Cob produced by the bacterial strain through fermentation has the functionfor degrading sodium alginate to prepare alginate oligosaccharide. The provided alginate lyase Aly-Cob can be widely used for the fields of agriculture, food, feed addition, medicine and alga processing.

Methods and compositions for releasing the nucleic acids from a variety of different types of microorganisms are provided. The method relies on a simplified lysis procedure that can be applied to many types of bacteria and fungal cells and is readily automated for high throughput screening methods. The method utilizes a high concentration of a chelating agent and a mixture of lysing enzymes to accomplish the disruption of microbial cell walls and allow the release of the nucleic acid.

This invention provides a method of detecting enzyme activity on a solid medium. The enzyme substrate has a chromogenic portion comprising a catechol residue, in which a derivitising moiety is linked to the aromatic ring of the catechol via a bond, and an enzyme cleavable group which is attached via an ester or ether linkage to the oxygen atom derived from a hydroxyl group of the catechol residue. If the enzyme substrate contacts an enzyme capable of cleaving the enzyme cleavable groups and the cleaved compound contacts a chelatable metal ion, a substantially non-diffusable coloured precipitate is formed.

The invention relates to the field of biological engineering, and in particular relates to lywallzyme (or catenase) of phage of staphylococcus aureus and an application thereof as an antibacterial material in preventing and treating bouine mastitis and controlling food pollution. The amino acid sequence of the lywallzyme of phage of staphylococcus aureus is SeqIDNO2. Zymin capable of efficiently killing staphylococcus aureus is developed by a bioengineering technique. The zymin can be used independently or with other agents to inactivate staphylococcus aureus specifically so as to provide a safe zymin source without toxic and side effects for preventing and treating bouine mastitis and controlling staphylococcus aureus pollution in food.

The invention relates to a lysis buffer mixture that is stable in storage for isolating nucleic acids from biological, preferably diagnostic samples. The mixture is preferably associated with an extraction control. The aim of the invention is to provide an improved nucleic acid extraction system, which is cost-effective, stable and easy to use, thus fulfilling the requirements of a modern nucleic acid extraction system and containing, among other things, extraction controls. The invention relates to a lysis buffer mixture for isolating nucleic acids, said mixture containing non chaotropic salts, a special selection of detergents, a defined quantity of at least one nucleic acid as an extraction control, optionally lytic enzymes, optionally carrier nucleic acids and optionally other additives.

The invention discloses phage lytic enzymes as well as a gene recombinant expression vector and application thereof. The phage lytic enzyme expression gene is a nucleotide sequence shown as SEQ ID NO:1, and the nucleotide sequence can express phage lytic enzymes in a host cell by constructing a recombinant vector. The phage lytic enzymes obtained through purification have relatively high in-vitroand in-vivo bactericidal activity, acts quickly, is safe and harmless to organisms, can well treat multi-drug-resistance shigella infection, and also has a certain inhibition effect on staphylococcusaureus, vibrio parahaemolyticus and other pathogenic bacteria. The phage lytic enzymes have good lytic thallus activity within the range of 28-42 DEG C, and due to the characteristics, the phage lytic enzymes have a wide prospect in the aspect of preparing drug-resistant infectious disease medicaments and can be used as a substitute or supplement of conventional antibiotics.

The invention discloses a rapid purification method for recombinant pyrolytic enzyme. According to the method, escherichia coli is used as an expression host of recombinant pyrolytic enzyme, the recombinant pyrolytic enzyme is subjected to inducible expression in host cells of escherichia coli, thalli are collected centrifugally, and escherichia coli is broken by heating to release expressed pyrolytic enzyme; meanwhile, host protein of escherichia coli is denatured, devitalized and precipitated at high temperature, which is favorable for purifying pyrolytic enzyme resisting to thermal denaturation; further, high-purity pyrolytic enzyme is obtained rapidly through the process of bacteria debris catching and protein concentration of pyrolytic enzyme through ultra-filtration. The rapid purification method is easy to operate; the escherichia coli protein thermal denaturation removal rate is high; the purification concentration efficiency is high; the method is adaptable to industrial production and marketing promotion and application.

The invention discloses a high-temperature-resistant Escherichia coli phage RDP-EC-20031 with the preservation number of CGMCC No.21406 and the gene length of 135812bp, which is discovered to have no virulence gene and lysogenic gene after sequencing the whole gene, and the gene of the phage contains a plurality of genes of lytic enzyme. The Escherichia coli phage RDP-EC-20031 provided by the invention has higher temperature tolerance and wider acid-base tolerance range, and is beneficial to industrial production and preparation; and the phage is found to have a lysis rate of 98% by lysis rate test and RTD test, and whenthe phage diluted by 107 times,the phage still has a good cracking effect on pathogenic bacteria. The phage has the characteristics of excellent performance and safety. The phage can be applied to preparation of medicines for preventing and / or treating diseases caused by escherichia coli.

The invention discloses a staphylococcus aureus bacteriophage lyase as well as a preparation method and application thereof, and relates to the technical field of bioengineering. The invention provides a staphylococcus aureus bacteriophage lyase LysSA2, and the amino acid sequence of the LysSA2 is Seq ID NO.1; and the invention provides an encoding gene of the staphylococcus aureus bacteriophage lyase LysSA2, and the nucleotide sequence of the encoding gene is Seq ID NO.2. The invention also provides a preparation method of the staphylococcus aureus bacteriophage lyase LysSA2. The method clones staphylococcus aureus bacteriophage SA2 to obtain the bacteriophage lyase LysSA2, and carries out induced expression on soluble lyase by adopting a prokaryotic expression mode. The bacteriophage lyase LysSA2 not only has a relatively strong bacteriostatic effect on staphylococcus, but also has a broad-spectrum bacteriostatic effect on staphylococcus from different sources, and lays a foundationfor treating and inhibiting staphylococcus diseases.

The invention discloses an alginate lyase encoding gene, and belongs to the technical field of biology. Alginate lyase is high in degrading activity and stable in property, enzyme activity reaches 65U / mg, 98% or more of initial enzyme activity is still kept after the alginate lyase is stored for 18 months at the temperature of 4 DEG C, the alginate lyase has high product specificity, and brown algae oligosaccharide trisaccharide can be specifically produced. The alginate lyase has important industrial application values and scientific research values.

The invention discloses a genetically engineered lysin with a function of killing staphylococcus and a preparation method. According to the preparation method, a lysin LysGH15 is subjected to genetically engineered modification to obtain a novel bacteriophage lysin LysGH15(+). The bacteriophage lysin LysGH15(+) shows stronger bactericidal activity and wider host spectrum, in comparison with a lysin before modification, can kill staphylococcus aureus, staphylococcus epidermidis, staphylococcus haemolyticus, staphylococcus hominis, staphylococcus saprophyticus, staphylococcus cohnii, fermented staphylococcus xylosus and the like, provides a potential novel medicine to prevention and treatment of diseases caused by staphylococcal infection, and has a good application value.

The invention provides an application of bacteriophage endolysins Lysep3 in preparation of a broad-spectrum antibacterial drug. The invention finds for the first time that the Lysep3 has a better inhibitory effect on gram-positive bacteria, and has an obvious inhibitory effect on a plurality of bacteria under the combined action of an outer membrane penetrant, and has lower hemolytic activity on mouse red blood cells and cytotoxicity to murine derived macrophages; and in addition, the natural bacteriophage endolysins Lysep3 is used as a template, a specific expression vector is constructed byoptimizing an endolysins Lysep3 gene sequence, so that the expression of the bacteriophage endolysins Lysep3 in an eukaryotic expression system (pichia pastoris) is realized, a protein purification system is established and perfected, the large-scale production of the bacteriophage endolysins Lysep3 can be realized, and the bacteriophage endolysins Lysep3 is used in the fields of antibacterial drugs and feed additives, and has important application value and broad market prospects.

The present invention in the field of oligosaccharide production provides a method of producing oligosaccharides of useful lengths without producing substantial amounts of monosaccharides and disaccharides (illustrated by FIG. 1). There is provided a method for producing an ingredient suitable for incorporation into a foodstuff, cosmetic, or nutraceutical, said ingredient comprising one or more oligosaccharides, wherein the oligosaccharides are produced in an enzymatic reaction, said enzymatic reaction comprising the step of contacting, in a solution or suspension, a polysaccharide-cleaving enzyme and a polysaccharide-containing feedstock, wherein said enzymatic reaction produces substantially no monosaccharides or disaccharides.

The present invention is generally related to engineered bacteriophages expressing antimicrobial peptides or lytic enzymes or fragments thereof for targeting a broad spectrum of bacterial hosts, and for the long-term suppression of bacterial phage resistance for reducing bacterial infections. In some embodiments, bacteriophages express antimicrobial peptides or antimicrobial polypeptides (e.g. phage lytic enzymes) which are secreted from the host bacteria, or alternatively released upon lysis of the bacterial host cell. Aspects of the present invention also relate to the use of the engineered bacteriophages for the reduction of bacterial infections, both in a subject or for bioremediation purposes, in clinical settings and wound healing.

The invention belongs to the technical field of biology, and in particular relates to helicobacter pylori bacteriophage endolysins and a preparation method thereof. A natural helicobacter pylori bacteriophage endolysin protein sequence is screened out in the invention; furthermore, easily permeable polycationic 9 peptide and hydrophobic small molecule peptide are respectively added into the N endof the protein sequence; fermentation-induced expression of an improved helicobacter pylori bacteriophage endolysin gene is carried out through an escherichia coli expression system; the expression quantity is high; a soluble expression method without deformation and renaturation is further provided; furthermore, a purification step is simple and convenient; the fact that the lysis ability of theimproved helicobacter pylori bacteriophage endolysins on a standard helicobacter pylori strain ATCC700392 is high is found through an in-vitro experiment; and material and theoretical basis can be provided for research on treatment of helicobacter pylori infection.

There is disclosed a method of engineering bacteriophages comprising: identifying a bacteriophage with only one attachment gene; isolating said bacteriophage; removing said attachment gene from the genome of said bacteriophage; and inserting a non-natural attachment gene into the genome of said bacteriophage wherein said non-natural attachment gene is specific for attaching to a selected bacteria.There is also disclosed a mutant bacteriophage comprising a heterologous nucleic acid sequence encoding a first specific attachment gene, the first specific attachment gene being different than an inactivated attachment gene and being specific for a selected bacteria. In another embodiment, there is disclosed a method of eliminating a microbial contaminant, the method comprising: obtaining one ormore lytic enzymes produced by a mutant bacteriophage; applying the one or more lytic enzymes to a bacterial contaminant, without prior infection of the bacterial contaminant with a bacteriophage, toeliminate the bacterial contaminant.

The invention relates to a microorganism bacterial strain and application thereof, and particularly relates to a photobacterium phosphoreum and application thereof, overcoming the defects that no report about photobacterium phosphoreum which is used in production of alginate lyase in the prior art is available, the alginate lyase produced by other bacterial strains has low output and is poor in storage stability, which greatly limits the use in industrial production. The photobacterium phosphoreum SRU-2 is preserved in China Center For Type Culture Collection on December 29, 2013 with a preservation number of CCTCC M2013723. The bacterial strain can be used for producing alginate lyase. The alginate lyase produced by the photobacterium phosphoreum has high output, and the alginate lyase liquor has good storage stability, so that the application range of the alginate lyase in industrial production is greatly enlarged.

The invention provides production and application of staphylococcus bacteriophage lysins LysK and antibacterial peptide Mersacidin fused protein. A nucleotide sequence which expresses the fused protein is shown in SEQ ID NO:1 as shown in the description, and an amino acid sequence of the fused protein is shown in SEQ ID NO:2 as shown in the description. According to production and application of the staphylococcus bacteriophage lysins LysK and antibacterial peptide Mersacidin fused protein, the fused protein is subjected to heterologous expression and purification, it is found through detection that the fused protein is stable, operative temperatures and pH values are wide in range, and the fused protein has specific sterilization effects on multiple kinds of staphylococcocci, including staphylococcus aureus. According to the provided antistaphylococcic fused protein, a safe protein preparation source is provided for control of staphylococcocci in food, especially dairy products, so that the provided antistaphylococcic fused protein has wide application prospects.

The invention provides a strain capable of degrading kelp. The strain is an isoptericola salitolerans ISO-49 strain, and the preservation number is CGMCC No.18975. The isoptericola salitolerans ISO-49provided by the invention can be used for fermenting the kelp to produce a seaweed liquid fertilizer. The strain ISO-49 used in the invention has very strong ability to produce alginate lyase, and can quickly destroy the structure of kelp cell walls to release nutrient substances of the kelp; the produced alginate lyase can decompose alginate polysaccharides in the kelp and make the alginate polysaccharides degraded into small-molecule substances that can be easily absorbed and utilized; and the yield of seaweed polysaccharides after fermentation is much higher than a yield of seaweed polysaccharides extracted by a general enzymatic method.

The invention relates to the technical field of microorganisms, in particular to arthrobacter pseudoarthrobacter PL-410 and application of the arthrobacter pseudoarthrobacter PL-410 in production of chondroitin lyase. The preservation number of the pseudoarthrobacter sp. PL-410 provided by the invention is CGMCC (China General Microbiological Culture Collection Center) No.22736, and the preservation number of the pseudoarthrobacter sp. PL-410 is CGMCC No.22736. The pseudoarthrobacter sp. PL-410 disclosed by the invention can be used for preparing chondroitin lyase (AC type), and the chondroitin lyase can be used for researching the structure of chondroitin sulfate and constructing a chondroitin sulfate oligosaccharide library, can also be applied to the field of medicines, and has a wide application prospect.

The invention discloses a strain of Bacillus and its application in industry. The strain screened rotten kelp picked up from the Yellow Sea and was preserved in the China Center for Type Culture Collection on April 28, 2019. The preservation number is: CCTCC No. .M 2019315, which can produce alginate lyase, and the enzymatic activity of the produced alginate lyase can adapt to a wide temperature range, between 30-45°C. The enzyme activity of the alginate lyase produced by the strain screened by the present invention is suitable for a wide temperature range and has certain advantages in the industry. The fermentation medium specially developed for the strain with the preservation number of CCTCC No.M 2019315 has sufficient nutrition. The strain can grow at a high density, so it produces more alginate lyase, has a stronger ability to degrade alginate, and has a higher yield of the product fucoidan oligosaccharide.

An improved, cost effective and shortened process of purification of the capsular polysaccharide of S. pneumoniae, Group B Streptococcus, H. influenzae, S. typhoid and N. meningitidis is disclosed. The process includes a cocktail of enzyme treatment, tangential flow filtration, and multimodal chromatography purification. For Gram-negative bacteria, endotoxin removal process involves endotrap HD resin. This shortened process achieved the purity required by WHO / EP / BP for the use in human vaccine preparation, with simple steps and higher yield as compared to conventional processes. The steps of the process avoid use of organic solvents such as, for example, alcohol, phenol, and ultracentrifugation that are otherwise expensive and time consuming to perform, and / or health hazards for commercial use. This process disclosed is also simple, efficient, non-toxic, easy to scale-up, and environmentally friendly.

The invention discloses a method for expressing anthrax bacteria gamma bacteriophage lysins. The invention connects an anthrax bacteria bacteriophage gene and pMEX9K, by adopting a pichia pastoris expression system and an expression vector pMEX9K (ZL02117906.9) with intellectual property right, to construct a new expression plasmid pMEXplyG; reservoir host Pichia pastoris GS115 undergoes electrotransformation after linearization; a producing strain G6 is obtained after random selection and pressure filtering, so that the gamma bacteriophage lysins are expressed in pichia pastoris G6 supernatant, and a expression product does not contain superfluous amino acid; and the gamma bacteriophage lysins with the purity of over 95 percent are obtained after centrifuging, ultrafiltering, ion-exchange chromatography, dewatering chromatography and gel chromatography. The anthrax bacteria gamma bacteriophage lysins have cleavage activity to anthrax bacteria and fungus gemma in sprouting, and can be used to develop anthrax decontaminating agents and anthrax medicines.

The invention provides alginate lyase and a gene and application thereof and further provides a method of cloning and expression of the alginate lyase in escherichia coli and yeast cells. A nucleotide sequence of the alginate lyase is shown as SEQ ID NO.1, and an amino acid sequence of the alginate lyase is shown as SEQ ID NO.3. A recombinant vector comprising the gene is formed by connection of the alginate lyase gene to escherichia coli plasmids or yeast plasmids. A cell comprising the gene is obtained by conversion of the recombinant vector. The cell of the alginate lyase gene comprises the nucleotide molecule or escherichia coli converted from the recombinant vector or comprises the nucleic acid molecule or Pichia pastoris converted from the recombinant vector.

The invention provides novel pharmacologic action of non-steroidal anti-in-flammatory medicines, i.e. the action for preventing and treating the diabetic encephalopathy. The non-steroidal anti-inflammatory medicines (ibuprofen) can be used for obviously improving the injury of the cognitive function caused by the diabetes mellitus, obviously increasing the protein expression and the mRNA (messenger ribose nucleic acid) level of intracerebral peroxisome proliferators-activated receptor gamma (PPAR gamma), obviously reducing the activity, the protein expression and the mRNA level of cerebral cortex beta amyloid precursor protein gamma-beta amyloid cleaving enzyme 1 (beta amyloid cleaving enzyme 1), the level of end-stage advanrced glycation end products (AGEs), and the protein expression of cyclo-oxygenase 2 (COX-2) and inducible nitric oxide synthase (iNOS), and obviously increasing the level of superoxide dismutase (SOD) reduced glutathione (GSH) in blood serum, etc. The non-steroidal anti-in-flammatory medicines can be used for preventing and treating the diabetic complication, i.e. the diabetic encephalopathy.

The invention relates to a preparation method and antibacterial application of plesiomonas shigelloides phage endolysin. The amino acid sequence of the plesiomonas shigelloides phage endolysin is Seq ID NO.2, and the plesiomonas shigelloides phage endolysin can be prepared into medicine inhibiting bacterial contamination or overgrowth and is applied as bactericide. According to the endolysin, amido bonds between saccharide and peptide of bacterial cell walls or linked bonds between amino acid residues in peptide are hydrolyzed to achieve lysis of host cells finally. The endolysin can act on a host specifically within a short action time and has a wider action spectrum than phage. The endolysin has the advantages of being efficient, specific, free of resistance and the like, and successful application of the endolysin has been reported in the food industry and the pharmaceutical industry at present.