A Klebsiella strain that efficiently utilizes crude glycerol to produce 1,3-propanediol
A Klebsiella, crude glycerol technology, applied in the field of microbial engineering, can solve the problem of less potential strains for industrial application and the like
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Embodiment 1
[0017] Embodiment 1: the species identification of bacterial strain
[0018] 1. Analysis of colony morphological characteristics
[0019] Streak inoculate a single colony of the strain on nutrient agar medium (peptone 10g / L, beef extract 3g / L, NaCl 5g / L, agar 15g / L, pH 7.0-7.2), and then place the plate upside down in a 30°C constant temperature incubator , cultured for 18-24 hours, and observed the shape of the colonies grown. attached figure 1 The colony showing the strain is light yellow and moist, small, round and convex, with neat edges and smooth surface. For Gram-negative bacteria.
[0020] The characteristics are generally as follows: Colony characteristics: the colony is slightly yellow, the surface is rough and wrinkled, opaque, round or irregular, and the edges are not neat; it is stained with Gram with a kit, and the bacteria are observed and photographed under an oil microscope. figure 2 It shows that the strains are rod-shaped, blue-purple in color, and are ...
Embodiment 2
[0028] Embodiment 2: Growth characteristics in the process of bacterial strain fermenting crude glycerol
[0029] The bacterial solution cultivated to the logarithmic phase was inoculated into the crude glycerol fermentation medium at an inoculum size of 5% (v / v), cultivated at 37° C. and 170 rpm for 12 hours, and samples were taken every 2 hours for cell growth analysis. Bacterial growth analysis adopts ultraviolet spectrophotometer to detect the absorbance value (OD600) of sample solution 600nm place, the result is as attached image 3 shown.
Embodiment 3
[0030] Example 3: Characteristics of Enzyme Activity Related to 1,3-Propanediol Synthesis During Strain Fermentation of Crude Glycerol
[0031]The bacterial solution cultivated to the logarithmic phase was inoculated into the crude glycerol fermentation medium with an inoculum size of 5% (v / v), cultivated at 37° C. for 12 hours at 170 rpm, and sampled once every 2 hours for glycerol dehydratase, 1 , 3-propanediol oxidoreductase and glycerol dehydrogenase activity detection. After the samples used for enzyme activity detection were centrifuged at 8000rpm for 5min, the cell pellet was washed twice with dipotassium hydrogen phosphate-potassium dihydrogen phosphate buffer (0.02M, pH 7.2) and then suspended, and the suspended cells were broken by an ultrasonic breaker , centrifuged at 12000 rpm, 4°C for 10 min, and saved the supernatant for subsequent enzyme activity detection. The determination method of glycerol dehydratase is as follows: in the total reaction system of 1.0ml, m...
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