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High-temperature Streptomyces thermoviolaceus and application of High-temperature Streptomyces thermoviolaceus in cellulose degradation

A technology of Streptomyces violaceum and cellulose, applied in the field of agricultural resources and environmental microorganisms, can solve problems such as unreported, unsuitable for large-scale industrial application, and small biomass.

Active Publication Date: 2019-02-19
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

The reported actinomycetes with cellulose degrading function mainly include Nocardia, Streptomyces, Thermoactinomyces, Micromonas, Thermoascomyces, etc. Slow speed, small biomass and weak degradation ability are not suitable for large-scale industrial application
[0004] Streptomyces hyperthermia ( Streptomyces thermoviolaceus ) is an aerobic Gram-positive bacterium, which can be used as a new source of antibiotic adjuvant streptazolin and molecular cloning of chitinase gene in existing reports. There are no reports on the application in the degradation of

Method used

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  • High-temperature Streptomyces thermoviolaceus and application of High-temperature Streptomyces thermoviolaceus in cellulose degradation
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  • High-temperature Streptomyces thermoviolaceus and application of High-temperature Streptomyces thermoviolaceus in cellulose degradation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Streptomyces violaceum St-GW2 strain improved CMC solid culture and Congo red staining experiment

[0033] For the composted samples of traditional Chinese medicine dregs in different sampling periods (from the compost of traditional Chinese medicine dregs in Donghai County, Lianyungang City, Jiangsu Province), take 5g and add them to 50 ml of sterilized ultrapure water, mix them evenly, centrifuge at 55°C and 150 rpm for 30 min, and take 1ml of the suspension Sequentially serially diluted, 100 µl of the diluted suspension was spread on the improved CMC solid medium, cultured in a 55°C incubator for 3 days, and the colonies on the improved CMC solid medium were streaked and purified, and the purified single strain was in Congo red staining was carried out after overnight culture on the improved CMC solid medium. After washing with 1 mol / L NaCl, there were transparent circles on the plate, which were possible high-temperature cellulose-degrading bacteria.

[00...

Embodiment 2

[0037] Example 2 Determination of Cellulase Activity of Streptomyces hyperthermicus Streptomyces violaceum St-GW2 strain

[0038] In order to measure the change curve of the enzyme activity of the strain, Streptomyces hyperthermicus St-GW2 was inoculated into the enzyme-producing fermentation medium, cultured at 55°C and 150 r / min for 14 days, and the strains were incubated at 1, 3, 5, 7, 9, On the 11th and 13th day, samples were taken for cellulase activity determination.

[0039] Enzyme activity detection method is as follows: CMCase measurement: Take 0.5ml of crude enzyme solution in a volumetric flask, add 1.5ml of 0.5mol / L, pH5.0 citric acid buffer solution containing 0.5% CMC-Na, and place in a water bath at 50°C Accurately act for 30 minutes, then add 1.5ml 3,5-dinitrosalicylic acid (DNS) solution, and immediately cool with running water after 5 minutes in boiling water bath. Measure its OD value at 540nm, and calculate the glucose content through the glucose standard ...

Embodiment 3

[0044] Example 3 Degradation ability of Streptomyces violaceum Streptomyces St-GW2 strain on filter paper cellulose

[0045] 1. Test tube filter paper degradation

[0046] The single colony of Streptomyces violaceum Streptomyces St-GW2 screened in Example 1 was inoculated into the improved test tube filter paper strip medium, and the filter paper strip without colonies was used as the control (K), and cultured under the same conditions as the initial screening temperature, Observe the ulceration of the filter paper strips.

[0047] The schematic diagram of the colony enrichment of strain St-GW2 in the test tube filter paper medium, the initiation of disintegration and the disintegration of the filter paper into a paste are shown in Figure 5 as shown, Figure 5 Among them, A, B, and C are pictures of test tubes cultured for 48h, 4d, and 8d in turn. It can be seen that at 48h, a large number of colonies are enriched and start to disintegrate ( Figure 5 A); at the 4th day, t...

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Abstract

The invention discloses high-temperature Streptomyces thermoviolaceus with the preservation number of CGMCC NO:16559 and application of the high-temperature Streptomyces thermoviolaceus in cellulose degradation. The Streptomyces thermoviolaceus belongs to high-temperature actinomycetes, the Streptomyces thermoviolaceus can quickly grows under the culture conditions of 55 DEG C, a lot of bacterialcolonies can be enriched within 24-48 hours, and the Streptomyces thermoviolaceus is easy to culture and preserve and has a good stable cellulose degradation function. By means of the Streptomyces thermoviolaceus, the filter paper degradation rate can reach 91.60+ / -0.020, and the straw degradation rate can reach 62.30%+ / -0.023. The CMC enzyme activity of the Streptomyces thermoviolaceus reaches the maximum on the fifth day and reaches 35.487 U / mL, and the Fpase enzyme activity reaches the maximum on the seventh day and reaches 62.383 U / mL. The Streptomyces thermoviolaceus can play an importantrole in promoting compost maturity, especially shortening the high temperature period and accordingly reducing the compost maturation cycle, industrial high-temperature degradation and treatment of solid cellulosic wastes are facilitated, the Streptomyces thermoviolaceus plays an important role in changing agricultural waste resources into valuable things, protection and optimization of the ecological environment are greatly facilitated, and therefore, the high-temperature Streptomyces thermoviolaceus has good resource and environment application values and market potential.

Description

technical field [0001] The invention relates to the fields of agricultural resources and environmental microorganisms, in particular to a Streptomyces hyperthermicus Streptomyces St-GW2 and its application in cellulose degradation. Background technique [0002] With the development of modern society and economy, the energy crisis and environmental problems are becoming more and more serious. The accumulation of a large number of solid wastes such as straw and domestic garbage has caused excessive environmental pressure. Cellulosic biomass can be solved and utilized, especially the The production of ethanol or advanced biofuels from agricultural waste such as straw, straw, and chaff can turn waste into wealth, which is one of the ways to solve future energy and environmental problems, and is of great significance. In the process of cellulose degradation, it is first necessary to effectively degrade and release the cellulose polysaccharides in the plant biomass, especially the...

Claims

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Application Information

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IPC IPC(8): C12N1/20C05F11/00C05F11/08C05F17/00C12R1/465
CPCC05F11/00C05F11/08C05F17/00C12N1/205C12R2001/465Y02W30/40
Inventor 杜婕马艳
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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