High-temperature resistance glucose isomerase, its coding gene and use

A technology of glucose isomerase and coding gene, which is applied in high temperature-resistant glucose isomerase and its coding gene and application field, and achieves the effect of good heat resistance

Inactive Publication Date: 2005-12-28
张力彬
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although scientists in many countries are interested in improving the heat resistance of glucose isomerase and have conducted a lot of research, see the literature: Journal of General Microbiology, 1993, 139: 1227-1234; Methods Enzymology 2001; 330: 215-24; Protein Engineering, 2000, 13: 259-65, but these studies are still difficult to produce fructose syrup with high concentration of fructose (such as greater than 42%) under the conditions of higher than the existing industrial conditions, that is, at a temperature above 60°C

Method used

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  • High-temperature resistance glucose isomerase, its coding gene and use

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Embodiment

[0073] 1) Gene amplification and plasmid construction

[0074] Design primers according to gene bank (L38994) gene sequence T1: 5'AGA TTA CCT AGG TACATA TGG CTG AAT TCT ATC CAG AAA TCC CGA AA 3' and T2: 5'ATT TCC GATGGC GCG CCT CAC CTC AGT TCC AGA ATG GTC TTC 3 '; The gene encoding glucose isomerase (TN) was amplified from Thermotoga neapolitana using primer pair T1 and T2.

[0075] Amplification reaction conditions were: 5 microliters of 10× buffer (Promega, USA), 4 microliters of 2.5 millimolar dNTPs (Promega, USA), 2 microliters of 10 micromolar primer T1, 2 microliters of 10 micromolar primer T2, 0.5 microliter 5U / microliter Tag DNA polymerase (Promega, USA), pick a little Thermotoga neapolitana bacteria with an inoculation loop, and then use sterile water to adjust the reaction volume to 50 microliters.

[0076] The PCR amplification reaction program was: 95°C for 3 minutes, 40 cycles: 95°C for 50 seconds, 50°C for 30 seconds, 72°C for 1 minute, and finally 72°C for 10 m...

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Abstract

High-temperature resistant dextrose isomerase, its coding gene and use are disclosed. The isomerase is characterized by having amino acid sequence in sequence table SEQ ID No:2 and base sequence in sequence table SEQ ID No:1, and deriving from Thermotoga neapolitana. It can be used for high fructose corn syrup production.

Description

technical field [0001] The present invention relates to glucose isomerase, specifically a kind of high temperature resistant (heat) glucose isomerase and its coding gene and application (in the aspect of fructose syrup production). Background technique [0002] Glucose isomerase, or xylose isomerase (Xylose isomerase) is an important industrial enzyme. In the food industry, glucose isomerase is used to prepare fructose syrup. When enzymatically producing fructose syrup, the higher the reaction temperature, the higher the fructose content in the product fructose syrup (Hartley, BS et al., Biochim Biophys Acta. 2000, 1543: 294-335.). [0003] At present, commercially used glucose isomerases are mainly obtained from Streptomyces, or Actinoplanes missouriensis, etc., but these glucose isomerases are not resistant to high temperature, at high temperature (such as higher than 60 ℃) easily deactivated. Therefore, at present, the industry has to prepare fructose syrup at 60°C, an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/92
Inventor 张力彬
Owner 张力彬
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