Lactobacillus brevis for producing feruloyl esterase
A technology of ferulic acid esterase and ferulic acid esterase enzyme, which can be applied to bacteria, microorganism-based methods, microorganisms, etc., can solve problems such as limiting the utilization and degradation of cellulose, and achieve the effect of improving digestibility and fermentation quality.
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Embodiment 1
[0027] Screening experiment of strains producing ferulic acid esterase.
[0028] 1) The lactic acid bacteria are activated 2-3 times, streaked on the MRS solid medium, and cultured at 37° C. for 2 days.
[0029] 2) Pick a single bacterium colony on the MRS solid medium without adding glucose and taking ethyl ferulate as the only carbon source, and add 0.3ml of ethyl ferulate solution to each plate (dissolve ethyl ferulate in Dimethylformamide, made into a solution with a mass volume ratio of 10%), cultivated at 30° C. for 72 hours, and observed whether a transparent bacterial circle appeared on the plate.
[0030] 3) Those that produce transparent bacterial circles can be preliminarily determined to produce ferulic acid esterase. A strain of Lactobacillus brevis FE3 produced a transparent circle.
Embodiment 2
[0032] Liquid culture enzyme production and enzyme activity assay.
[0033] 1) The strain Lactobacillus brevis FE3 after primary screening was cultured overnight in MRS liquid medium at 37°C, centrifuged at 3000rpm for 5min, and the bacteria were collected.
[0034] 2) The cells were washed three times with 0.85% physiological saline, and resuspended in deionized water.
[0035] 3) Take the bacterial suspension (determinate bacterial count 1×10 9 cfu / ml) was inoculated into the medium containing methyl ferulate (add 5ml methyl ferulate (1%W / V dimethylformamide) in the 100ml medium, and the bacterial classification inoculum was 2% (V / V) for fermentation. The fermentation condition is 37° C., 48 hours, and the shaking frequency is 120 rpm.
[0036] 4) Take 9ml of fermentation broth into a 10ml centrifuge tube, centrifuge at 10,000rpm for 10min, and the supernatant is ferulic acid esterase crude enzyme solution, which is used for enzyme activity determination.
[0037] 5) Tak...
Embodiment 3
[0046] Physicochemical properties of FAE-producing Lactobacillus brevis.
[0047] 1) Sugar fermentation, arginine ammonia production, nitrate reduction and glucose gas production: Bacterial trace biochemical reaction tubes are used; the strains are activated 2-3 times, streaked for 2-3 days, and a single colony is picked with an inoculation loop in the biochemical tube , sealed with sterilized glycerin, and incubated at 37°C for 2-3 days to observe the discoloration.
[0048] Table 2 The utilization ability of lactic acid bacteria of the present invention to sugar source
[0049]
[0050] Note: "+" means that the sugar is used; "—" means that the sugar is not used.
[0051]2) Growth characteristics of strains under different temperature conditions: after 2-3 times of lactic acid bacteria activation, culture overnight at 37°C, the number of bacteria is 1×10 8 cfu / ml, inoculate into sterilized MRS liquid medium with 3% inoculum amount, shake well, seal with parafilm, and se...
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