Streptomyces griseorubens beta-1, 4-glucosidase and coding gene and application thereof
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A technology of Streptomyces grisea and glucosidase, which is applied in the field of genetic engineering and modern enzyme engineering, can solve the problems of low content and low activity, achieve high-efficiency expression, convenient purification, and ensure biological activity
Inactive Publication Date: 2017-03-08
SHANGHAI JIAO TONG UNIV
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However, related studies have shown that β-1,4-glucosidase has a low content and low activity, and is the rate-limiting enzyme in the process of cellulose degradation
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[0037] Below in conjunction with specific embodiment, further illustrate the present invention. It should be understood that these examples are only used to illustrate the present invention, not to limit the protection scope of the present invention. Improvements and adjustments made by those skilled in the art according to the present invention in practical applications still belong to the protection scope of the present invention.
[0038] 1. Genome extraction and sequencing of Streptomyces grisea
[0039] Inoculate the spore suspension of Streptomyces grisea into LB liquid medium, culture at 37°C and 150rpm for 3 days, then collect 2.0mL of the bacterial liquid, centrifuge at 12000rpm for 2min, discard the supernatant, collect the bacterial precipitate, and then Add 180 μL lysozyme (20 mg / mL) and 20 μL EDTA solution (0.5M, pH 8.0), treat at 37°C for 45 minutes, add 4 μL RNase A (100 mg / mL), shake and mix for 15 seconds, leave at room temperature for 5 minutes, and then fol...
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Abstract
The present invention discloses a Streptomyces griseorubens beta-1, 4-glucosidase and a coding gene and application thereof. A protease gene is cloned from Streptomyces griseorubens, and a nucleotide sequence is shown as SEQ ID NO.1. An amino acid sequence of the above encoded beta-1, 4-glucosidase is SEQ ID NO.2. The invention also relates to application of the beta-1, 4-glucosidase and the encoded gene of the beta-1, 4-glucosidase in hydrolyzation of polysaccharide substances. The addition of a soluble label is convenient for subsequent purifying while keeping the protein activity. The enzyme is an acidic and high temperature resistant glucoside hydrolase, and its activity is less affected by the concentration of salt ions. The Streptomyces griseorubens beta-1, 4-glucosidase has strong tolerance to a plurality of metal ions and enzyme activity inhibitors, thereby having important meaning in the real production and application.
Description
technical field [0001] The invention belongs to the technical field of genetic engineering and modern enzyme engineering, and relates to a Streptomyces grisea β-1,4-glucosidase and its coding gene and application, in particular to a β-1,4-glucosidase and Its coding gene sequence, the recombinant expression vector containing the coding gene sequence, the transgenic recombinant bacteria and the application of the β-1,4-glucosidase. Background technique [0002] Cellulase is composed of exo-β-1,4-glucanase, endo-β-1,4-glucanase and β-1,4-glucosidase, in which β-1,4-glucoside Enzymes are key enzymes in sugar metabolism, which can hydrolyze glycosidic bonds and convert crystalline cellulose, amorphous cellulose, and soluble cellobiose into glucose, and are widely found in archaea, bacteria, and eukaryotes. Under the synergistic action of exo-β-1,4-glucanase, endo-β-1,4-glucanase and β-1,4-glucosidase, cellulose is completely transformed into glucose. However, relevant studies h...
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