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Cellulase-producing marine bacteria HN B15, microbial preparation thereof and degradation method of natural cellulose

A technology of natural cellulose and marine microorganisms, applied in microorganism-based methods, methods of using microorganisms, biochemical equipment and methods, etc., can solve problems such as low tolerance of enzyme-producing strains, complex ecological structure and instability

Active Publication Date: 2021-04-16
HAINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Existing cellulase-producing strains are often selected from terrestrial environments with rich nutrients and complex ecological structures
Due to the obvious differences in the environment and climate between my country's inland areas and Hainan Island, the obtained enzyme-producing strains are usually not highly tolerant and unstable, resulting in ineffective use of many aquatic probiotics in Hainan Island
Existing common aquatic probiotics are not suitable for utilizing cellulose

Method used

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  • Cellulase-producing marine bacteria HN B15, microbial preparation thereof and degradation method of natural cellulose
  • Cellulase-producing marine bacteria HN B15, microbial preparation thereof and degradation method of natural cellulose
  • Cellulase-producing marine bacteria HN B15, microbial preparation thereof and degradation method of natural cellulose

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] 1. Collect samples

[0042] On December 23, 2018, the seagrass bed ecosystem in the mangrove area of ​​Huiwen Town, Wenchang City (19°28′11.66″N, 110°47′41.22″E)( figure 1 ) to collect sediment mud samples below 3cm near the seagrass bed. The collected mud samples were placed in sterile zipper plastic bags and stored in an ice box at 4°C.

[0043] 2. Separation and purification of microorganisms

[0044] Within 24 hours of collecting the sample, weigh 0.5 g of the mud sample with a sterile spoon and suspend it in a sterile saline solution (0.85% NaCl), stir evenly, and use this as the stock solution to serially dilute the water sample with a sterile saline solution. To obtain dilutions of 1:10, 1:100 and 1:1000, 100 μL of each diluted sample was uniformly spread on 2216E seawater agar medium (0.5% trypsin, 0.1% yeast, 0.01‰ iron phosphate and 1.7% agar ) and cultured in a 30°C incubator for 18 hours. Investigate colony morphology on agar plates, including shape, siz...

Embodiment 2

[0054] 1. Determination of filter paper enzyme activity

[0055] Take the culture of 2216E liquid grown overnight, centrifuge at 4°C, 5000rpm, 15min; take 0.5mL of the supernatant enzyme solution, add it to 1mL of acetic acid-sodium acetate buffer solution with a pH of 4.8 and 0.1mol / L, and then add 50± One piece of 0.5mg filter paper (1cm×6cm), insulated at 50°C for 1 hour, (preheated for 5 minutes); add 3mL of DNS chromogenic solution, put it into boiling water for 10 minutes, and cool it under running water at 540nm Measure the absorbance; at the same time, use the inactivated enzyme solution boiled at 100°C for 10 minutes as a control to subtract the background; find out the corresponding glucose content from the glucose standard curve according to the absorbance, and calculate the enzyme activity value according to the grams of glucose produced. Filter paper enzyme activity was calculated according to the following formula:

[0056] X=(W×N×1000) / (T×M)

[0057] X: Enzyme...

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Abstract

The invention relates to the technical field of microbial preparations, in particular to marine bacteria for producing cellulase, a microbial preparation of the marine bacteria and a degradation method of natural cellulose. The preservation number of the marine bacteria for producing the cellulase is CCTCC No: M20191040. Cellulase-producing spore bacteria suitable for tropical prawn mariculture environments such as Hainan are screened from a Hainan hay bed ecosystem in China, and basic evaluation of application of the cellulase-producing spore bacteria is carried out; therefore, a foundation is laid for developing a marine microbial preparation efficiently utilizing cheap cellulose and constructing a prawn mariculture mode of artificially adding cheap artificial carbon sources such as bagasse.

Description

technical field [0001] The invention relates to the technical field of microbial preparations, in particular to cellulase-producing marine bacteria, microbial preparations and natural cellulose degradation methods. Background technique [0002] According to statistics in 2018, my country's total aquaculture output is greater than 50 million tons, making it a fishery country whose total aquatic products exceed the total catch in the world. Although high-density farming can bring high returns, the accumulation of high-protein bait and excrement will aggravate the deterioration of the breeding water. According to research reports, residual bait and manure are the main sources of elements such as nitrogen and phosphorus during the breeding process, and the relative lack of carbon elements leads to deterioration of water bodies. The recently emerging biofloc culture mode regulates the carbon-nitrogen ratio of the water body by adding additional carbon sources. However, the cost ...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/22C02F3/34C12R1/07C02F103/20
Inventor 谢珍玉吴金萍徐雪妮龙昊章翔黄爱优
Owner HAINAN UNIVERSITY
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