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Anaerobic alginate decomposing bacterium and application thereof

A technology of alginate decomposing bacteria and alginate lyase, which is applied in the direction of bacteria, lyase, microorganisms, etc., can solve the problems of limiting the large-scale application of alginate oligosaccharides, the lack of specificity in acid hydrolysis, and the inability to effectively control oligosaccharides. Achieve high enzyme production, avoid bacterial contamination, and shorten the time period

Inactive Publication Date: 2014-01-22
KUNMING UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The acid hydrolysis method needs to consume a large amount of high-concentration acids such as hydrochloric acid, and the hydrolysis waste residues cause serious environmental pollution. The acid hydrolysis is not specific, and the size of oligosaccharides and the composition of mannuronic acid and guluronic acid cannot be effectively controlled. These shortcomings are all Limiting the large-scale application of alginate oligosaccharides

Method used

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  • Anaerobic alginate decomposing bacterium and application thereof
  • Anaerobic alginate decomposing bacterium and application thereof
  • Anaerobic alginate decomposing bacterium and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1: Screening and isolation of anaerobic strain SH-52

[0041] The seabed sediment collected from the coast of Shantou City, Guangdong Province was diluted 1:10 (W / V), and inoculated into a 250ml anaerobic reagent bottle containing 100ml of modified marine medium (the carbon source was 10g / L sodium alginate ), cultured at 30°C, and transferred to fresh anaerobic modified marine medium after two days of cultivation, cultured to absorbance OD=0.8, transferred to fresh test tube medium, and inoculated three times (inoculum size: 2%). The culture was serially diluted (10 -1 、10 -2 、10 -3 、10 -4 、10 -5 、10 -6 ), using a Hunter roller tube (the culture medium is a modified marine medium, the carbon source is 10g / L sodium alginate, plus 1.5% agar) to separate, and pick a single colony to inoculate in the modified marine carbon source of sodium alginate In the culture medium, a strain capable of utilizing sodium alginate was screened out, numbered SH-52, and named ...

Embodiment 2

[0042] Embodiment 2: Physiological and biochemical detection

[0043] 1. Insert the bacterial strain obtained in Example 1 into the improved marine culture medium with a sodium alginate concentration of 10g / L at an inoculum size of 2%, at 20°C, 25°C, 30°C, 35°C, and 40°C respectively , 45°C, and 50°C culture, the measured temperature growth range of bacteria is 20°C to 45°C, and the optimum temperature is 30°C (see image 3 ).

[0044] 2. Configure the modified marine medium with pH gradients, the pH gradients are 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9. The measured pH range of bacterial growth is 6 to 8.5, and the optimum growth pH is 7.5 (see Figure 4 ).

[0045] 3. Configure the NaCl concentration gradient to improve the marine medium, and the NaCl concentration gradient is 0%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% (w / v), the measured growth range of bacteria on the NaCl concentration gradient medium is 0% to 9%, and the optimum growth concentration is 2% (see Figu...

Embodiment 3

[0049] Example 3: Preparation of Alginate Lyase Crude Enzyme Liquid

[0050] Insert the bacterial strain obtained in Example 1 into the anaerobic marine culture medium with a sodium alginate concentration of 10 g / L at an inoculum size of 2%, culture it statically at 30°C for 36 hours, and centrifuge at 10,000g for 10 minutes to collect the bacterial cells. Resuspend the bacteria in 50mM Tris-HCl buffer solution with pH 7.5, crush the bacteria with an ultrasonic breaker with a power of 200W, sonicate for 3s, pause for 3s, run for 8min, then centrifuge at 12000g for 15min, collect the supernatant, and obtain crude Enzyme solution.

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Abstract

The invention discloses an anaerobic alginate decomposing bacterium Marinicatena alginatilytica SH-52. The strain is preserved in the China Center for Type Culture Collection and the preservation number of the strain is CCTCCNO: M2013073. The bacterial strain of the decomposing bacterium disclosed by the invention is an anaerobe capable of effectively degrading alginate and can be applied to the production of alginate lyase; and the decomposing bacterium is short in time period, high in enzyme yield, simple to collect collection; the operation is convenient. The strain can be put into industrial implementation easily.

Description

technical field [0001] The invention relates to an anaerobic alginic acid decomposing bacterium and its application, belonging to the field of seaweed chemical industry. Background technique [0002] With the gradual development of marine resources, polysaccharides in seaweed, especially alginic acid, are widely used in various industrial productions. The alginic acid oligosaccharides obtained by the degradation of alginic acid have different biological activities due to different molecular weights, different M / G, and different structures, making them widely used in many fields such as medicine, food, and agriculture. In the field of medicine, sulfated oligosaccharide SPMG (sulfated polymannuroguluronate) can inhibit HIV-1 from entering cells [Geng M, Li F, Xin X, et al(2003). The potential molecular targets of marine sulfated polymannuroguluronate interfering with HIV-1 entry. Interaction between SPMG and HIV-1 rgp120 and CD4 molecule. Antiviral Res, 59(2):127-135]. Algin...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N9/88C12R1/01
Inventor 伊日布斯钱龙尚淑梅
Owner KUNMING UNIV OF SCI & TECH
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