Biogenic amine degrading bacterium for improving poor fermented flavor of shrimp paste and application thereof
A technology of unfavorable flavor and biogenic amines, which is applied in the field of microorganisms to achieve the effects of reducing accumulation, improving unfavorable flavor and improving quality
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Embodiment 1
[0031] Example 1 Screening of biogenic amine-degrading bacteria
[0032]Biogenic amine-degrading bacteria were screened from the strains isolated from grasshopper shrimp paste in the early stage, and a total of 442 halophilic bacteria were isolated. After 16S rRNA gene identification, 103 bacteria belonging to 46 genera were obtained (Sang, X. , Ma, X.X., Hao, H.S, Bi, J.R., Zhang, G.L & Hou, H.M. (2020). Evaluation of biogenic amines and microbial composition in the Chinese traditional fermented food grasshoppersub shrimp paste. LWT, 134, 109979.). Screen decarboxylase-negative bacteria from these 103 kinds of bacteria, use an inoculation loop to pick a single colony in LB solid medium, and then in a liquid decarboxylase medium containing histidine, tyrosine, lysine and ornithine (Ingredients: tryptone 5g, yeast powder 5g, beef extract 5g, glucose 0.5g, sodium chloride 2.5g, Tween 80 1mL, magnesium sulfate 0.2g, manganese sulfate 0.05g, ferrous sulfate 0.04g, ammonium citrate...
Embodiment 2
[0033] Morphological observation of embodiment 2 pantothenic mycobacterium (Virgibacillus pantothenicus)
[0034] The Gram staining method includes four steps of primary staining, mordant staining, decolorization, and counterstaining. The specific method is as follows: the smear of Virgibacillus pantothenicus was fixed, stained with ammonium oxalate crystal violet for 1 min, rinsed with distilled water, and then washed with iodine. Dye for about 1 minute, wash with water, absorb water with absorbent paper, add a few drops of 95% alcohol, and shake gently to decolorize, wash with water after 20 seconds, absorb water, dye with safranin staining solution for 1 minute, rinse with distilled water , dried, and examined under a microscope.
[0035] The result is as figure 1 As shown, Mycobacillus pantothenica is rod-shaped in shape, and its physiological characteristics are Gram-positive.
Embodiment 3
[0036] Example 3 Degradation of biogenic amine by Virgibacillus pantothenticus
[0037] Inoculate Virgibacillus pantothenicus (Virgibacillus pantothenicus) in 100mL LB liquid medium with an inoculum amount of 1% (v / v), culture at 37°C and 150r / min for 24h (bacteria liquid), and collect bacteria by centrifugation at 4°C and 4000g for 10min body, washed twice with 0.05mol / L pH=7.0 phosphate buffer, then resuspended the bacteria to 20mL salt containing four biogenic amines (500mg / L each for histamine, tyramine, putrescine and cadaverine concentrations) ( Sodium chloride, add) in the phosphate buffer saline (0.05mol / L, pH=7.0) of mass concentration 5%, adjust the bacterial suspension concentration to be OD 600nm =0.8, and then cultivated at 37°C and 150r / min for 48h. The control group was 20 mL of 5% phosphate buffer (0.05 mol / L, pH=7.0) containing 500 mg / L of each of the four biogenic amines (sodium chloride, plus) and cultured for 48 hours under the same conditions. After the ...
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