Marine streptomyces viridochromogenes strain for producing alkali-tolerant and salt-tolerant xylanase and application of marine streptomyces viridochromogenes strain

A technology of Streptomyces chromogenes and xylanase, applied in the biological field, can solve the problems of high pressure on production and environmental protection, and achieve the effects of increased yield, high enzyme activity, and broad application prospects

Inactive Publication Date: 2013-03-13
DALIAN UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the commonly used methods for industrial degradation of xylan are acid method, alkali method and enzymatic method. Among them, acid method and alkali method are widely used, but the acid-base hydrolyzate contains more toxic substances, which can inhibit the microbial fermentation in the later stage. ; In addition, the use of acid or alkali to degrade xylan has a great pressure on the production of environmental protection, so it is restricted by the country. Therefore, from the perspective of long-term production effects, enzymatic degradation of xylan will be the main method for future xylan degradation. Way

Method used

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  • Marine streptomyces viridochromogenes strain for producing alkali-tolerant and salt-tolerant xylanase and application of marine streptomyces viridochromogenes strain
  • Marine streptomyces viridochromogenes strain for producing alkali-tolerant and salt-tolerant xylanase and application of marine streptomyces viridochromogenes strain
  • Marine streptomyces viridochromogenes strain for producing alkali-tolerant and salt-tolerant xylanase and application of marine streptomyces viridochromogenes strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Embodiment 1: the acquisition of bacterial strain

[0052] The sea mud samples from Dalian Xiaoping Island were enriched and cultured, diluted according to the routine, and spread on the xylan plate, cultured at 30°C for 5-6 days, and the colonies produced by the transparent circle were picked and placed on the xylan solid medium Streak isolation and repeat the streak isolation process for 3 rounds to purify the strain. The strain that secretes xylanase is screened by this method.

[0053] The bacterial strain of the present invention is cultured on a Bennett plate at 30° C. for 7 days, and the bacterial colony is circular, with a diameter of about 3-4 cm, the base is dark green, the surface of the bacterial colony protrudes, produces white-gray spores, and releases a large amount of black pigment. Its optimum pH is 7.0-9.0, and its optimum temperature is 30°C.

Embodiment 2

[0054] Example 2: Extraction of Streptomyces viridans Genomic DNA and the acquisition of its 16SrDNA

[0055] Extraction of Streptomyces viridans Genomic DNA:

[0056] 1) The strain was inoculated in 50 mL of TSB liquid medium, and cultured at 30° C. at a rotational speed of 170 rpm for 72 hours. At this time, it was in the logarithmic growth phase. Collect 1mL of fermentation broth, centrifuge at 8000r / min for 5min, discard the supernatant, wash the cells with washing solution, and repeat the washing once after centrifugation.

[0057] 2) To the washed cells, add 100 μL of lysate to suspend the cells, boil at 100° C. for 15 min, and then cool to room temperature.

[0058] 3) Add 0.5ml of the extract solution preheated at 65°C and shake slightly.

[0059] 4) Add an equal volume of Tris-saturated phenol / chloroform solution, shake vigorously for 1 min, centrifuge at 10,000 r / min for 5 min, carefully absorb the supernatant, and transfer to a clean centrifuge tube.

[0060] 5) ...

Embodiment 3

[0072] Example 3: Oligotrophic tolerance, pH tolerance and NaCl tolerance of Streptomyces viridans chromogenes

[0073] Dilute Bennett's solid medium by 1, 2, 3, and 5 times to determine the oligotrophic tolerance of Streptomyces viridans; adjust the pH of Bennett's solid medium to 5, 7, 9, and 11 to determine the strain's tolerance pH tolerance; add 0%, 1%, 2%, 3% NaCl to Bennett solid medium to determine the NaCl tolerance of the strain. Take 50 μL spore solution (concentration is about 10 6 cells / mL) were evenly spread on each of the above-mentioned plates, cultured at 30°C for 6 days, and the growth status of the colonies was observed. Results (see Figure 1-3 ) shows that the bacterial strain grows well on the Bennett plate diluted 5 times, indicating its better oligotrophic tolerance; the bacterial strain grows poorly on the plate of pH 5, and can still grow normally on the plate of pH 11, indicating The bacterial strain of the present invention has strong alkali resi...

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Abstract

A marine streptomyces viridochromogenes strain for producing alkali-tolerant and salt-tolerant xylanase and application of the marine streptomyces viridochromogenes strain belong to the field of biotechnology. The strain is collected in the China General Microbiological Culture Collection Center, and the collection number is 5565. The strain can normally grow under the extreme alkaline environment of pH11, shows a good oligotrophy-tolerant characteristic, can degrade a variety of hemicellulose-containing biomasses including corn straws, rice straws, Jerusalem artichoke straws, kelp fibers and the like, and has a broad application prospect in utilizing cheap biomass resources to produce bio-based chemicals. The strain can produce xylanase under optimum conditions, and the enzymatic activity of the xylanase can be as high as 68.9U / ml. The produced xylanase has the following properties: the optimum pH value is 6.0, the optimum temperature is 70 DEG C, the optimum substrate is beech xylan, and the produced xylanase does not have cellulase activity, has high pH stability and thermal stability, and can tolerate high-concentration NaCl. As an enzyme preparation coming from the ocean, the xylanase can be widely used in papermaking, food, feed, wine making, energy and other industries.

Description

technical field [0001] The invention relates to a marine viridans chromogenic streptomyces (M11) strain for producing alkali-resistant and salt-tolerant xylanase and application thereof, which is especially suitable for large-scale industrial production of alkali-resistant and salt-resistant xylanase, and belongs to the field of biotechnology. Background technique [0002] The use of cheap biomass resources to produce high value-added products is an important research content of biotechnology. Xylan is an abundant renewable resource in nature, and is the most representative hemicellulose component, accounting for 1 / 3-1 / 2 of hemicellulose, and is the most abundant polysaccharide in nature except cellulose . Therefore, how to utilize xylan has become a hot research topic at home and abroad. [0003] Xylan is a polymer composed of D-xylose backbone (linked by β-1,4 bonds) and L-arabinose branches (linked by α-1,2 and α-1,3), and lignin It is connected with cellulose by 4-O-m...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12R1/465
Inventor 赵心清刘卓白凤武
Owner DALIAN UNIV OF TECH
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