Death bacillus vallismortis for producing spore laccase and application of death bacillus vallismortis

A technology of bacillus, spore laccase, applied in the field of biology

Inactive Publication Date: 2012-10-31
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are few studies on bacterial laccases, and more bacterial laccases with high-efficiency decolorization ability need to be further explored and studied.

Method used

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  • Death bacillus vallismortis for producing spore laccase and application of death bacillus vallismortis
  • Death bacillus vallismortis for producing spore laccase and application of death bacillus vallismortis
  • Death bacillus vallismortis for producing spore laccase and application of death bacillus vallismortis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 The acquisition of spore-producing laccase dead Bacillus vallismortis fmb-103

[0043] Add 10g of soil samples from printing and dyeing factories to 250mL sterilized enrichment medium with glass beads under aseptic conditions (enrichment medium component: 1% (W / V) tryptone ; 0.5% (W / V) yeast powder; 1% (W / V) NaCl; final concentration 0.2mmol / L CuSO 4 ). Shake at 200 rpm for 24-48 hours at 37°C to enrich the culture medium. After the cultivation, centrifuge at 10,000rpm and 4°C for 10min, and obtain the supernatant for gradient dilution to obtain 10 -1 、10 -2 、10 -3 、10 -4 、10 -5 、10 -6 、10 -7 、10 -8 、10 -9 the dilution factor. Add 100 μL of bacterial solution for each gradient dropwise on the nutrient agar plate (nutrient agar plate components: 10.0g / L peptone; 3.0g / L beef powder; 5.0g / L sodium chloride; 15.0g / L agar; 1000ml distilled water; pH Value 7.3 ± 0.1) after coating, cultured at 37°C for 2-4 days. Take a plate containing a single colony, ...

Embodiment 2

[0044] Example 2 Identification of spore-producing laccase dead Bacillus vallismortis fmb-103 strain

[0045] (1) Morphological identification of strains

[0046] Bacillus death valley fmb-103 (CGMCC No.6198) strain was inoculated on nutrient agar medium and cultured at 30°C for 24 hours. The characteristic observation results are shown in Table 1. The colony morphology is shown in figure 1 , Gram staining results see figure 2 .

[0047] Table 1 Observation results of culture characteristics of bacterial strain fmb-103

[0048]

[0049] (2) Study on the biological characteristics of the strain

[0050] 1) Determination of acid and alkali resistance

[0051] Bacillus death valley fmb-103 (CGMCC No.6198) strain was inoculated in the In nutrient agar liquid medium (10.0g / L peptone; 3.0g / L beef powder; 5.0g / L sodium chloride; 1000ml distilled water; pH value 7.3±0.1), culture at 30°C, and compare with uninoculated control every 12h Tube comparison, visual inspection of t...

Embodiment 3

[0082] Example 3: Fermentation of Bacillus vallismortis fmb-103 to obtain spore laccase

[0083] (1) Pick the nutrient agar plate ((10.0g / L peptone; 3.0g / L beef powder; 5.0g / L sodium chloride; 30.0g / L agar; 1000ml distilled water; pH value 7.3±0.1)) Bacillus death valley fmb-103 (CGMCC No.?) single colony, in 100mL BPY seed liquid medium (beef extract 5g / L, peptone 10g / L, yeast extract 5g / L, glucose 10g / L, NaCl 5g / L L), 37°C, 180rpm, cultured for 12-16 hours.

[0084] (2) Transfer the cultured seed solution to the fermentation medium (corn flour 4.65g / L, soybean flour 5.88g / L, MgSO 4 2.53g / L, MnSO 4 0.1g / L, K 2 HPO 4 0.3g / L, FeSO 4 0.5g / L, pH 7.0), 37°C, 180rpm, ferment for 48 hours.

[0085] (3) Collect the fermented liquid, and remove the insoluble matter in the fermented liquid by suction filtration;

[0086] (4) Suction filtrate, centrifuge at 9000rpm for 30min;

[0087] (5) After centrifugation, discard the supernatant, resuspend the precipitate with deionized wat...

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Abstract

The invention belongs to the field of biotechnology, and relates to a death bacillus vallismortis bacterial strain fmb-103 for producing spore laccase. The death bacillus vallismortis bacterial strain fmb-103 for producing the spore laccase is preserved in the China general microbiological culture collection center (CGMCC), the preservation date is on June 8, 2012, and the preservation number is CGMCC No.6198. The death bacillus vallismortis bacterial strain fmb-103 the preservation number of which is CGMCC No.6198 can produce the spore laccase, and can be applied in degradation triphenylmethane dyestuff.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a spore-forming laccase-producing Bacillus vallismortis strain fmb-103. Background technique [0002] The dye industry is an important industry in the national economy. The dyes it produces are widely used in various fields such as textiles, papermaking, food, leather, cosmetics, coatings, paints, inks, and electronics. There are tens of thousands of dyes, and about 10% to 20% of dyes will directly enter the environment with wastewater during production and use. Most dyes are toxic and refractory organic substances with strong chemical stability and heavy metals, which are irritating, allergenic, acutely toxic, carcinogenic, teratogenic, and mutagenic, and directly endanger human health; in addition, dyes in wastewater can Absorbs light and reduces the transparency of water, which is not conducive to the growth of aquatic organisms and microorganisms, and reduces the self-purification...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/02C02F3/34C12R1/07
Inventor 陆兆新张充吕凤霞刁含文别小妹王昱沣赵海珍
Owner NANJING AGRICULTURAL UNIVERSITY
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