51 results about "Thermostable enzymes" patented technology

The present invention is directed to a process for amplifying any target nucleic acid sequence contained in a nucleic acid or mixture thereof using a thermostable enzyme. The process comprises treating separate complementary strands of the nucleic acid with a molar excess of two oligonucleotide primers, extending the primers with a thermostable enzyme to form complementary primer extension products which act as templates for synthesizing the desired nucleic acid sequence, and detecting the sequence so amplified. The steps of the reaction can be repeated as often as desired and involve temperature cycling to effect hybridization, promotion of activity of the enzyme, and denaturation of the hybrids formed.

The present invention is directed to a process for amplifying any target nucleic acid sequence contained in a nucleic acid or mixture thereof using a thermostable enzyme. The process comprises treating separate complementary strands of the nucleic acid with a molar excess of two oligonucleotide primers, extending the primers with a thermostable enzyme to form complementary primer extension products which act as templates for synthesizing the desired nucleic acid sequence, and detecting the sequence so amplified. The steps of the reaction can be repeated as often as desired and involve temperature cycling to effect hybridization, promotion of activity of the enzyme, and denaturation of the hybrids formed.

The present invention provides a method for obtaining thermostable enzymes. The present invention also provides variants of DNA polymerase I from Thermus aquaticus. The present invention further provides methods of identifying mutant DNA polymerases having enhanced catalytic activity. The present invention also provides polynucleotides, expression systems, and host cells encoding the mutant DNA polymerases. Still further, the present invention provides a method to carry out reverse transcriptase-polymerase chain reaction (RT-PCR) and kits to facilitate the same.

This invention relates to selection and application of thermophil denitriding Bacillales,of which conservation number is CGMCC-1228. Initial strain is from water of oil field. The strain is obtained by selection, inoculation, and domestication with good thermal stability. It belongs to soil Bacillales and can be used in fermented industry, which needs thermally stable enzyme. It grows well in the oil field and can degrade paraffin and decrease viscosity to improve liquidity. All of this can increase recovery and carriage efficiently. It can be used to control pollution and prepare surfactant.

The present invention provides a method for obtaining thermostable enzymes. The present invention also provides variants of DNA polymerase I from Thermus aquaticus. The present invention further provides methods of identifying mutant DNA polymerases having enhanced catalytic activity. The present invention also provides polynucleotides, expression systems, and host cells encoding the mutant DNA polymerases. Still further, the present invention provides a method to carry out reverse transcriptase-polymerase chain reaction (RT-PCR) and kits to facilitate the same.

The present invention relates to methods and compositions for providing purified thermostable enzymes, particularly thermostable DNA polymerases, that are free of exogenous detergents. The present invention also provides methods for providing such purified thermostable DNA polymerases to assays in an active form by adding one or more detergents. The present invention further provides compositions and kits comprising purified thermostable DNA polymerases for use in a variety of applications, including amplification and sequencing of nucleic acids.

A purified thermostable enzyme is derived form the thermophilic archaebacterium Archaeoglobus fulgidus. The enzyme can be native or recombinant, is stable under PCR conditions and exhibits double strand specific exonuclease activity. It is a 3′-5′ exonuclease and cleaves to produce 5′-mononucleotides. Thermostable exonucleases are useful in many recombinant DNA techniques, in combination with a thermostable DNA polymerase like Taq especially for nucleic acid amplification by the polymerase chain reaction (PCR).

A purified thermostable enzyme is derived form the thermophilic archaebacterium Archaeoglobus fulgidus. The enzyme can be native or recombinant, is stable under PCR conditions and exhibits double strand specific exonuclease activity. It is a 3′-5′ exonuclease and cleaves to produce 5′-mononucleotides. Thermostable exonucleases are useful in many recombinant DNA techniques, in combination with a thermostable DNA polymerase like Taq especially for nucleic acid amplification by the polymerase chain reaction (PCR).

The invention relates to strains of Talaromyces emersonii which are thermostable and encode thermostable enzymes. The enzymes retain activity at temperatures above 550 C. These strains and enzymes find use in a variety of processes from waste reduction to the production of novel food ingredients and the production of bio-fuels.

Described herein is a thermostable enzyme capable of efficient ligation of two oligomers at high temperature. The embodiments herein have led to the development of an optimized ligation step used in library preparation for sequencing reactions.

The present invention is directed to compositions comprising mixtures of reagents, including thermostable enzymes (e.g., thermostable DNA polymerases), buffers, cofactors and other components, suitable for immediate use in nucleic acid amplification or sequencing techniques without dilution or addition of further components. The compositions contain no stabilizing agents (e.g., glycerol or serum albumin) and unexpectedly maintain activity for extended periods of time upon storage at temperatures above freezing. These compositions are useful, alone or in the form of kits, for nucleic acid amplification (e.g., by the Polymerase Chain Reaction) and sequencing (e.g., by dideoxy or “Sanger” sequencing), or for any procedure utilizing thermostable DNA polymerases in a variety of medical, forensic and agricultural applications. In particular, the compositions and methods are useful for amplifying and sequencing nucleic acid molecules that are larger than about 7 kilobases in size.

The present invention relates to methods and compositions for providing purified thermostable enzymes, particularly thermostable DNA polymerases, that are free of exogenous detergents. The present invention also provides methods for providing such purified thermostable DNA polymerases to assays in an active form by adding one or more detergents. The present invention further provides compositions and kits comprising purified thermostable DNA polymerases for use in a variety of applications, including amplification and sequencing of nucleic acids.

The invention relates to strains of Talaromyces emersonii which are thermostable and encode thermostable enzymes. The enzymes retain activity at temperatures above 550C. These strains and enzymes find use in a variety of processes from waste reduction to the production of novel food ingredients and the production of bio-fuels.

The present invention provides thermostable enzymes, such as DNA polymerases and restriction endonucleases, that are substantially free from contamination with nucleic acids. The invention also provides methods for the production of these enzymes, and kits comprising these enzymes which may be used in amplifying or sequencing nucleic acid molecules, including through use of the polymerase chain reaction (PCR).

The disclosure provides thermostable enzymes isolated from Caldicellulosiruptor bescii and fragments thereof useful for the degradation of cellulose and / or hemicellulose, including thermostable cellulases and hemicellulases. The disclosure further provides nucleic acids encoding the thermostable enzymes of the disclosure. The disclosure also provides methods for the conversion of cellulose and hemicellulose into fermentable sugars using thermostable enzymes of the disclosure. The disclosure also provides enzyme cocktails containing multiple enzymes disclosed herein. The enzymes can be used to release sugars present in cellulose or hemicellulose for subsequent fermentation to produce value-added products.

The invention provides a strain of Geobacillus thermodenitrificans as well as the screening method and the uses thereof. The strain was deposited as the number CGMCC-1228 in the China General Microbiological Culture Collection Center of the China Committee of Culture Collection for Microorganisms. The strain was obtained by primary screening, re-screening, inoculation, domestication and propagation using the bacteria in the aqueous samples from the strata of oil field as the original screening strain. The strain screened in the invention belongs to the genus Geobacillus, which can tolerate a high temperature, and has good thermostability, which can be applied to the industrial production which need the condition of thermostable enzymes, such as fermentation, etc. With its ability of growing well in the oil-reservoir environment, degrading alkanes, decreasing viscosity and increasing fluidity of crude oil, the strain is able to remarkably enhance the yield of oil and improve the transporting efficiency of oil. Additionally, with its good ability of degrading oil, it can be developed for the treatment and clearing of the material such as the oil contaminated water, etc., so it can be useful in protection of the environment. This strain also has the ability of decreasing the surface tension of substances, which can be applied to the industry of biosurfactants preparation.

The present invention relates to the field of thermostable enzymes, methods for their inactivation, and related uses, kits and reagents. In particular, the invention relates to a method for reversible inactivation of thermophilic enzymes by chemical modification under aqueous or non-aqueous conditions.

In one aspect, the invention provides a purified thermostable enzyme derived from the archael bacterium AEPII 1a. In one aspect, the enzyme has a molecular weight of about 60.9 kilodaltons and has a cellulase activity. The enzyme can be produced from native or recombinant host cells, and can be used to aid in the digestion of cellulose. The invention also provides polypeptides having endoglucanase activity having homology to SEQ ID NO:2.

The invention aims to provide a high-efficiency expression and purification method for family-10 xylanase Aus Xyn10A thermostability modified and reconstructed mutant enzyme. According to a comparison result of protein sequences of the Aus Xyn10A (derived from Aspergillus usamii E001) and thermostable xylanase (derived from Thermotoga maritima), the C terminal repeat of thermostable enzyme is used to the C terminal of the Aus Xyn10A by a gene engineering method, and the obtained mutant enzyme is named Aus Xyn10A'. Experimental results prove that the optimum temperature of the mutant enzyme is obviously improved, and the thermostable enzyme serving as a thermostable enzyme preparation has high industrial production potential and economic value.

In a method for amplifying the specific nucleic acid sequence, a highly specific amplification which has low possibility of non-specific hybridization can be carried out. Highly stable reagents, the activities of which do not decrease in case of supply and storage, are also provided. Thermostable enzymes are used as RNA dependent DNA polymerase, DNA dependent DNA polymerase, DNA dependent RNA polymerase and ribonuclease H, which are required for the amplification system based on replicated RNA. Especially, it is preferable that a thermostable enzyme derived from Thermus thermophilus which has RNA dependent DNA polymerase activity, DNA dependent DNA polymerase activity and ribonuclease H activity, and thermostable DNA dependent RNA polymerase are used together. By this method, inactivation of enzymes are prevented by using thermostable enzymes, and the amplification can be carried out without sequential addition of enzymes.

The invention relates to a method for treating a cellulosic grey fabric comprising the following steps: (a) a pretreatment step in which, in the presence of water, at a temperature of 60-100 DEG C, the fabric is contacted with a thermostable enzyme which degrades starch; and (b) an integrated desizing and scouring step in which, in the presence of water, at a temperature of 70 DEG C at the most, the fabric as obtained in step (a) is contacted with an enzyme which degrades a polymeric component of the primary cell wall of cotton and an enzyme which degrades starch. The invention also relates to the use of fabric as obtained using the method of the invention for manufacturing textile products. The invention also provides fabric manufactured using the method of the invention. The invention further relates to textile products manufactured from fabric treated using the method of the invention.

It is intended to improve a method of adding a non-thermostable enzyme to a candy base material. In the method of the invention, a non-thermostable enzyme is added to a candy base material, and the material is stirred so that a substantially uniform mixed dough-like mixture is obtained within 5 minutes from the addition to obtain the mixed dough-like mixture. Then, the mixed dough-like mixture is cooled on a cooling plate and the mixed dough-like mixture is shaped. Preferably, a main component of the candy base material is selected from the group consisting of reduced palatinose, maltitol, reduced starch syrup, and xylitol.

The present invention provides reversibly modified thermostable enzyme compositions and methods for making the same. The present invention also provides methods of using the reversibly modified thermostable enzyme compositions, as well as kits and systems comprising the reversibly modified thermostable enzymes.

The present invention relates to improved methods and reagents for the production of 1,3-propanediol. In particular, the present invention provides novel thermophilic organisms and thermostable enzymes capable of catalyzing the fermentation of glycerol to 1,3-propanediol. The present invention also relates to methods of isolating such thermophilic organisms, methods of cloning polynucleotides that encode such enzymes, polynucleotides encoding such enzymes, and methods of using such enzymes and organisms for the production of 1,3-propanediol.

The sensitivity and specificity of polynucleotide synthesis is increased by protecting the 3'-end of an oligonucleotide used as a primer in the synthesis of the polynucleotide. Protection of the 3'-end of an oligonucleotide prevents non-specific chain elongation. Removal of blocking group an elevated temperature, using a thermostable enzyme, permits template-specific polynucleotide synthesis. The present invention also provides oligonucleotides with a 3' end protected by a blocking group and a thermostable enzyme capable of removing the blocking group at an elevated temperature. The compositions and methods of the invention are very useful in a variety of techniques for DNA / RNA amplification and analysis, including medical genetics research and diagnosis, pathogen detection, forensic, and animal and plant genetics applications, among others.

The present invention is directed to compositions comprising mixtures of reagents, including thermostable enzymes (e.g., thermostable DNA polymerases), buffers, cofactors and other components, suitable for immediate use in nucleic acid amplification or sequencing techniques without dilution or addition of further components. The compositions contain no stablizing agents (e.g., glycerol or serum albumin) and unexpectedly maintain activity for extended periods of time upon storage at temperatures above freezing. These compositions are useful, alone or in the form of kits, for nucleic acid amplification (e.g., by the Polymerase Chain Reaction) and sequencing (e.g., by dideoxy or “Sanger” sequencing), or for any procedure utilizing thermostable DNA polymerases in a variety of medical, forensic and agricultural applications. In particular, the compositions and methods are useful for amplifying and sequencing nucleic acid molecules that are larger than about 7 kilobases in size.