Strain producing alginate lyase and cellulase, and application of strain in kelp fermentation
A kelp and strain technology, applied in the biological field, can solve the problems of polluted environment, high cost of enzyme preparations, low degradation yield, etc., and achieve the effect of purifying water quality in aquaculture ponds and quick release.
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Embodiment 1
[0035] Embodiment 1: Screening and identification of Shewanella japonica NJ16-3
[0036] The steps of screening bacteria are as follows: figure 1 shown.
[0037] Step 1: First, collect sea mud from healthy sea cucumber breeding ponds in different locations along the coast of Rongcheng. Take 10 grams of different sea mud samples, add them to 90 mL of sterile physiological saline liquid medium, shake at room temperature for 2 hours at 150 r / min, and use the dilution coating method to separate bacterial strains. Take 0.1 mL of each sample and directly coat Put it on 2216E medium (Difco, product number: 212185), place it upside down at 30°C and culture it for 5 days, pick a single colony grown on the plate, streak and purify on the 2216E medium plate, and isolate a total of 121 strains of bacteria.
[0038] Step 2: Primary screening of strains producing alginate lyase
[0039] The isolated marine bacteria were inoculated on the alginate lyase-producing medium plate, cultured in...
Embodiment 2
[0071] Embodiment 2: Hemolysis experiment of Shewanella japonica NJ16-3
[0072] Whether it has the ability to produce hemolysin is an important screening method for eliminating pathogenic bacteria and retaining probiotics. Inoculate the strain NJ16-3 on a goat blood plate, incubate in a 30°C incubator for 24 hours, and then observe it. If a hemolytic ring is formed around the colony, it indicates that the bacteria has hemolytic ability and is a potential pathogenic bacterium; if no hemolytic ring occurs, it indicates that the bacteria No hemolytic ability.
[0073] The hemolysis test of strain NJ16-3 was negative, indicating that strain NJ16-3 could be used as a potential aquatic probiotic.
Embodiment 3
[0074] Embodiment 3: Shewanella japonica NJ16-3 degrades kelp block experiment
[0075]The bacterial strain NJ16-3 was inoculated in the kelp degradation liquid seed medium at a culture temperature of 30°C, placed on a shaker and cultivated at a speed of 150r / min for 48 hours, and inserted into the kelp block degradation liquid medium at 10% of the inoculum size at a temperature of 30°C. ℃, 150r / min, cultured for 3 days, and observed the degradation of kelp pieces.
[0076] Kelp degradation liquid seed medium: sodium alginate, 10g; sodium carboxymethylcellulose, 5g; peptone, 5g; yeast extract, 1g; sea salt, 10g; distilled water 1000mL; .
[0077] A composition of the kelp block degradation liquid medium is as follows: 20 g of kelp blocks (each about 10 cm in length and about 4 cm in width); 2 g of sea salt, 200 mL of distilled water, and sterilized at 121° C. for 20 min.
[0078] On the first day and the third day of fermentation, the results of degradation of kelp block are...
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