A kind of Pseudomonas betae producing keratinase and its application
A technology of pseudomonas and keratinase, applied in the field of microorganisms, can solve the problems of difficult direct utilization, energy and environmental pollution, difficult biodegradation, etc., and achieve the effect of considerable application prospects.
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Embodiment 1
[0025] Embodiment 1: Isolation and cultivation of keratinase-producing bacteria
[0026] Place the collected soil samples in sterile distilled water and mix them evenly, let stand to get the supernatant, and carry out gradient dilution (dilution to 10 -6 、10 -7 、10 -8 ), were respectively spread on the milk agar plate medium, and cultured upside down in a 37°C incubator for 24h. Select a single colony that can produce a transparent degradation circle, inoculate it in an inorganic liquid medium containing a feather of about 0.5 g, and cultivate it at 37° C. with shaking at 180 rpm for 72 hours. The culture fluid of strains with obvious feather degradation was selected and inoculated into a new inorganic liquid medium for re-screening culture.
[0027] Inoculate the strains obtained by re-screening into a 250mL Erlenmeyer flask containing a complete feather and 50mL of inorganic liquid medium, culture at 35°C and 200rpm with shaking for 48h, observe the degradation of feather...
Embodiment 2
[0030] Example 2: Identification of keratinase-producing bacteria
Embodiment 2A
[0031] Embodiment 2A) morphological characteristics
[0032] The isolated and purified target strain D1 was streaked and inoculated on milk agar plate medium, and cultured at 37°C for 24 hours. Visually inspect the colonies for shape, color, surface texture, and colony edges. The morphology of bacteria was observed by optical microscope and transmission electron microscope. Refer to the "Common Bacteria System Identification Manual" to identify the physiological and biochemical characteristics of the strains and make a preliminary classification.
[0033] The shape of the target colony under the transmission electron microscope is as follows: figure 1 shown. The colony of the target strain D1 was cultured on the milk agar plate medium for 24 hours to produce a transparent degradation circle, the colony was a yellow single colony with smooth edges; Gram staining was negative; short-chain rods, no spores, no capsule, and flagella .
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