Low-temperature amylase strain, low-temperature amylase, and production therefor
A technology of low-temperature amylase and production method, which is applied in the field of producing the enzyme and amylase, and can solve problems such as difficulty in separation
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Embodiment 1
[0073] Embodiment 1: the cultivation of low-temperature amylase-producing bacteria (LA140)
[0074] The low-temperature amylase-producing bacteria LA140 of the present invention is inoculated in broth medium containing soluble starch, wherein soluble starch 2%, beef extract 1%, peptone 1%, NaCl 0.5%, agar 3g, water 100mL, pH7, 0.1 MPa steam sterilization for 30 minutes. Cultivate at about 20°C for 2-3 days, add 0.4 mol / L I+2KI solution to develop color. Then an obvious transparent circle is formed around the colony of the low-temperature amylase-producing bacteria of the present invention.
Embodiment 2
[0075] Embodiment 2: the fermentation culture-I of low-temperature amylase producing bacterium (LA140 bacterial strain)
[0076] Inoculate the slant strain of the low-temperature amylase-producing bacteria of the present invention into the fermentation medium. Among them, peptone: 10g, corn flour: 15g, yeast extract: 2g, beef extract: 10g, sodium chloride: 1g, calcium chloride: 2g, ferrous sulfate: 0.1g, magnesium sulfate: 0.5g, water: 1000mL, pH7 .0. 30mL per 250ml Erlenmeyer bottle, 0.1MPa steam sterilization for 30 minutes. The culture conditions are: 20° C., 200 rpm, 36 hours of fermentation, and the enzyme activity reaches 28 U / mL.
Embodiment 3
[0077] Embodiment 3: Fermentation culture-II of low-temperature amylase-producing bacteria (LA140 bacterial strain)
[0078] The low-temperature amylase-producing bacteria LA140 was inoculated in broth peptone medium, in which 1% beef extract, 1% peptone, 0.5% NaCl, 1% soluble starch, pH 7.0, 0.1MPa steam sterilization for 25 minutes. After culturing for 36 hours at 20° C. and 150 rpm, the culture was added to the fermentation medium at a ratio of 15%.
[0079] The components of the fermentation medium are: peptone: 10g, corn flour: 15g, yeast extract: 2g, beef extract: 10g, sodium chloride: 1g, calcium chloride: 2g, ferrous sulfate: 0.1g, magnesium sulfate: 0.5g, Water: 1000mL. 0.1MPa steam sterilization for 25 minutes. 500ml Erlenmeyer bottle liquid 100mL. Shake culture at 20° C. and 200 rpm for 60 hours. After culturing, the crude enzyme solution of low-temperature amylase is obtained by centrifugal separation. The low-temperature amylase activity of the obtained amyla...
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