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Bacillus licheniformis, method for preparing flocculant from it and application of flocculant

A technology of Bacillus licheniformis and flocculant, applied in the field of microorganisms, can solve the problems of poor soil air permeability, increased sludge hardness, neurotoxicity and carcinogenicity, etc., and achieves the effects of wide growth temperature range, high flocculation activity and low production cost.

Active Publication Date: 2019-07-23
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI +2
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0005] Polyacrylamide (PAM) is commonly used in sludge conditioning and dehydration in the field of sludge treatment. However, the disadvantage of using PAM for dehydration is that the residual monomer in PAM, acrylamide, is neurotoxic and carcinogenic, and PAM is harmful to the environment. Degradation by physical and chemical methods will also produce acrylamide monomer; on the other hand, the PAM content in the sludge is too high (12-50mg / g absolute dry sludge), which will increase the hardness of the sludge, which is not conducive to subsequent treatment. Utilization will cause soil compaction and poor soil air permeability, which is not conducive to the growth of crops

Method used

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  • Bacillus licheniformis, method for preparing flocculant from it and application of flocculant
  • Bacillus licheniformis, method for preparing flocculant from it and application of flocculant
  • Bacillus licheniformis, method for preparing flocculant from it and application of flocculant

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Experimental program
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Embodiment 1

[0030] Embodiment 1, isolation and screening of Bacillus licheniformis (Bacillus licheniformis) LZ-1

[0031]The strain of the present invention was isolated and purified from the activated sludge of a wood pulping sewage treatment plant in Jiaozuo, Henan Province in 2012. The isolation of bacterial strains is done by diluting 1mL of activated sludge 10 times, then taking 0.2mL of the diluted solution and spreading it on a solid medium plate for separation. After culturing at 37°C for 3 days, pick a single colony and streak it several times on the same medium plate. , until pure species were obtained by microscopic examination.

[0032] The isolated pure strain was planted on the solid plate of the selection medium, and cultured at 37°C for 2 days. Pour 1% methyl red indicator reagent on the surface of the solid plate, react for 30 minutes, then add 1mol / L NaCl solution for decolorization, after reacting for 30 minutes, pour out the liquid, observe the hydrolysis circle aroun...

Embodiment 2

[0036] Embodiment 2, the morphology and physiological and biochemical characteristics of Bacillus licheniformis (Bacillus licheniformis) LZ-1

[0037] (1) Strain morphology observation:

[0038] Colony shape: Dilute the liquid culture of strain LZ-1 on a solid medium plate and culture it for 2 days. The colony is flat, round or nearly round, milky white, opaque, and about 2mm in diameter.

[0039] Morphological characteristics: use optical microscope and scanning electron microscope to observe the morphology of bacterial cells. The cells are rod-shaped (see figure 1 ), the size is about 1.5~2.0×0.5μm. The cells of the strain LZ-1 produced spores, the cysts were slightly enlarged, and there were no paraspore crystals.

[0040] (2) Physiological and biochemical characteristics of the strain:

[0041] Growth characteristics: The strain has the characteristics of wide growth temperature range and alkali resistance, and can grow under the conditions of 28°C-50°C and pH 8-11.

...

Embodiment 3

[0049] Embodiment 3, 16S rRNA gene sequence PCR amplification and sequence analysis of Bacillus licheniformis LZ-1

[0050] The strain LZ-1 was inoculated in the basal medium, and after 3 days of culture, the bacteria were collected by centrifugation, washed with buffer, and lysozyme was added to break the cells, and the genomic DNA was extracted by phenol: chloroform extraction, and the obtained DNA was used as a template , the 16S rRNA gene of strain LZ-1 was amplified by bacterial forward primer 27-F (5'-AGAGTTTGATCCTGGCTCAG-3') and reverse primer 1492-R (5'-GGTTACCTTGTTACGACTT-3'). PCR reaction system (100μl): 10μl of 10×Taq enzyme buffer, 25mmol / L MgCl 2 6μl, 10mmol / L dNTP 2μl, Taq DNase 2.5U, 30pmol / L upstream and downstream primers 2μl each, template DNA 2μg, distilled water to make up to 100μl. The PCR reaction conditions were: 95°C for 5min, 95°C for 1min, 58°C for 5min, 72°C for 1.5min, 30 cycles, 72°C for 10min, and storage at 4°C. The amplified products were sepa...

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Abstract

The invention relates to bacillus licheniformis, a method for preparing a flocculating agent from the bacillus licheniformis and an application of the flocculating agent. The strain is bacillus licheniformis LZ-1 which is preserved in China General Microbiological Culture Collection Center on November 06, 2012 with preservation number of CGMCC No.6782. With the application of the bacillus licheniformis LZ-1 as well as a production method provided by the invention, a fermentation broth, which is high in flocculating activity, is obtained under the condition that an initial pH value is alkaline, and high-molecular-weight polysaccharide, which is 0.8-3.2*10<7>Da, serves as a major flocculating active ingredient. The polysaccharide flocculating agent provided by the invention has the advantages of being high in flocculating activity, low in dosage, low in production cost and the like, and the flocculating agent has a flocculating conditioning effect on sludge. The bacillus licheniformis LZ-1 fermentation broth or a product, which is extracted from the fermentation broth and contains the polysaccharide flocculating agent, can be applied to deep dewatering by sludge plate-frame pressure filtration in a mode of replacing a polyacrylamide flocculating agent, so that sewage treatment plant concentrated sludge, which is 96-98% in moisture content, is further dewatered into a sludge cake which is lower than 60% in moisture content and the size of the sludge is diminished to be 1 / 20-1 / 10 of an original size.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a strain of bacillus licheniformis, a method for preparing a flocculant therefrom and the application of the flocculant. Background technique [0002] Microbial flocculants are metabolites with flocculation activity produced by microorganisms, mainly including polymer compounds such as mucopolysaccharides, glycoproteins, proteins, and bacteria with flocculation activity. Among them, there are many research reports on microbial polysaccharide flocculants. Microbial flocculant is a new, high-efficiency, non-toxic water treatment agent with biodegradability and safety. It can overcome the defects of inorganic flocculants and synthetic organic polymer flocculants in terms of safety and environmental pollution, and there are many types , a wide range of sources, less affected by seasonal changes. [0003] The existing microbial flocculant-producing bacteria have a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P19/04C02F11/147C12R1/10
Inventor 刘双江朱晓宇罗明芳冯权樊华肖娟娟蒋志坚缪强李雄伟宋联王瑞章旭胡科宏
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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