Serratia sp. A5 and uses thereof
A technology of Serratia and bacterial agent, which is applied to Serratia A5 and its use, and the application field of Serratia A5 in the degradation of kitchen waste, can solve the problem of a single variety of catalytic substrates, etc. Anaerobic fermentation hydrolysis and biogas production efficiency, improving hydrolysis efficiency, and releasing inhibitory effects
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Embodiment 1
[0019] Example 1 Serratia A5 Isolation and Preservation
[0020] Serratia A5 was isolated from soil by gradient dilution method.
[0021] The specific method is: Weigh 10 g (soil) soil from the place polluted by leftovers and leftovers around the cafeteria of the Hebei Academy of Sciences, add it to a triangular flask filled with 90 mL sterile water and glass beads, and shake it on a shaker for 30 min ( 150 r / min), serially diluted to make 10 -5 、10 -6 Dilute the suspension, draw 0.1 mL in each of them, add to the lipase primary screening medium (plus Tween-80) plate, spread evenly, and repeat 3 times for each treatment. Place the above petri dish at 25°C for constant temperature culture and transfer the single colony with large halo and fast growth to the slant of LB test tube, and store at 4°C.
[0022] Rescreening was carried out by enzyme activity assay. The isolated strains were placed in LB liquid medium and cultured in an incubator at 25 °C for 48-72 h. The ferme...
Embodiment 2
[0033] Example 2 Serratia A5 Enzyme Activity Assay
[0034] Serratia marcescens ( Serratia marcescens ), Serratia liquefaction ( Serratia liqufaciens ), Serratia odorifera ( Serratia odorifera ), Serratia rubrum ( Serratia rubidaea ) were respectively connected to LB liquid medium, cultured on a shaker at 30 °C for 48 h, the fermentation broth was centrifuged at 4 °C and 8000 r / min for 10 min, and the supernatant was taken to determine the lipase activity.
[0035] The determination methods are respectively adopted: National Standard GB / T 23535-2009.
[0036] Enzyme activity definition: 1 g1 mL liquid enzyme (or solid enzyme powder), under certain temperature and pH conditions (this example uses 40 ℃, pH 7.0), 1 min to hydrolyze the substrate (this patent uses olive oil as the substrate) Produce 1 umol of titratable fatty acid, that is, 1 enzyme activity unit, expressed in "U / mL (U / g)".
[0037]It can be seen from Table 1 that the lipase activity of Serratia A5 was ...
Embodiment 3~7
[0040] Example 3-7 Application of Serratia A5 in anaerobic fermentation of kitchen waste
[0041] Put the kitchen waste into the triangular flask first, set up 6 treatments, 5 experimental groups, insert Serratia A5 bacterial liquid at 5% to 20%, and the total number of bacteria in the bacterial liquid is ≥10 9 Individual / mL; 1 blank control group, no Serratia A5 was added. Shaker shaking culture for 13-24 hours, the oil in the experimental group completely disappeared, while the oil in the control group did not decrease.
[0042] Then a 5 L full-mix anaerobic fermentation device was used, and the above groups of kitchen waste were used as raw materials. The inoculum came from the normal fermentation biogas digester, and the inoculum amount was 25-30% (volume ratio), and the final anaerobic fermentation liquid TS was 5-5. 8% (TS, also known as dry matter concentration, refers to a certain amount of fermentation feed liquid placed in an oven at 105 ℃, dried to constant weig...
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