4 results about "Thermostable enzymes" patented technology

The application discloses a kind of high thermal stability glutamine transaminase mutant and its method of expression in streptomyces, belong to enzyme engineering field.The high thermal stability glutamine transaminase mutant provided by the application can efficiently catalyze casein crosslinking at high temperature, realizes the heterologous efficient expression of glutamine transaminase.Rational mutation is carried out to key amino acid site of glutamine transaminase, and the mutant strain with significantly improved thermal stability is obtained.Among them, the optimal mutant strain FRAPD-TGm2A3 has a half-life of up to 537.91 min at 60 DEG C.The mutant can still efficiently catalyze the crosslinking reaction of casein at high temperature of 85 DEG C.The present application not only lays a solid research foundation for the modification of thermal stability of glutamine transaminase, but also provides a generalizable strategy framework for the rational modification of other thermostable enzymes, and has important industrial application value and academic reference significance.

Provided herein are secretory compositions / cocktails derived from a novel, extremophilic fungal species cultured, isolated, and characterized herein. Said novel, extremophilic fungal species may be exploited to produce secretory compositions / cocktails which comprise a unique mixture of thermo-tolerant, stable enzymes capable of digesting cellular and genetic material in its surroundings under extreme temperatures. Such secretory compositions / cocktails may be useful as cleaning and sterilization solutions against biofilm, urine, yeast, bacterial, or viral contamination. Other uses include, but are not limited to, urea removal, fertilizer wastewaters, wastewater reclamation for life support systems in space, pH control, source of ammonia and carbon dioxide, starch digestion, biofuel production, as pharmaceuticals, and medical treatments such as wounds, skin disorders, or nail disorders.

This invention discloses a strategy for rapid detection of isoniazid resistance in Mycobacterium tuberculosis based on multiplex polymerase chain reaction (Multiplex PCR) using local rTth DNA polymerase and interpretation of amplification patterns. This strategy utilizes a specific primer combination that allows simultaneous amplification of the control fragment and the target fragment of the katG gene codon 315 mutation in a single PCR reaction. The amplification products are analyzed by 1% (w / v) agarose gel electrophoresis and interpreted based on the resulting DNA banding pattern to distinguish sensitive isolates, isolates carrying the katG315 mutation causing isoniazid resistance, and invalid test results. The use of local rTth DNA polymerase provides an alternative thermostable enzyme to support national diagnostic raw material independence without changing the detection principle.This strategy allows for rapid identification of katG315 mutations associated with isoniazid resistance without the need for DNA sequencing or further molecular analysis. The invention provides a simple, rapid, specific, easily interpretable, and cost-effective molecular diagnostic method to support the early detection of isoniazid-resistant tuberculosis.

The disclosure provides a thermostable enzyme-linked biosensor expression cassette comprising a nucleic acid comprising a nucleotide sequence encoding a β-glucosidase reporter. The enzyme-linked biosensor expression cassette of the disclosure comprises a nucleic acid comprising a nucleotide sequence encoding (i) a transcription factor, (ii) a promoter, (iii) a terminator, and (iv) a cloning site for a gene of interest. The disclosure further provides novel variants of β-glucosidase that function as the reporter enzyme and exhibit superior properties (e.g., without limitation, pH stability and thermal stability) compared to existing β-glucosidase, providing improved biosensor expression cassettes.