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- W946251671 abstract "Publisher Summary This chapter discusses the determination of mannitol dehydrogenase (crystalline) from Lactobacillus brevis . A spectrophotometric assay, based on the oxidation of reduced diphosphopyridine nucleotide (DPNH) by fructose is employed. The oxidation of DPNH is followed by the change in absorbance at 340 mμ. In the course of purification procedure, obtain a lyophilized culture of Lactobacillus brevis (ATCC 367) from the American Type Culture Collection. Grow the cultures in liquid medium by incubation at 32° without aeration. For preparation of large amounts of cells for enzyme purification, inoculate 10 ml of a 24-hour liquid culture into 1 liter of fresh medium. This is used 24 hours later as inoculum for 16 liters of medium in a 20-liter bottle. The purified enzyme is most stable in solution at pH 6.0 in a 0.02 M sodium acetate buffer containing 10 -3 M mercaptoethanol. Such solutions can be stored for weeks at –16°. The enzyme solutions lose no activity when dialyzed at 3° against oxygen-free water containing 2 × 10 -4 M mercaptoethanol for a period of 18 hours." @default.
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- W946251671 date "1966-01-01" @default.
- W946251671 modified "2023-09-27" @default.
- W946251671 title "[28] Mannitol dehydrogenase (crystalline) from Lactobacillus brevis" @default.
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- W946251671 doi "https://doi.org/10.1016/0076-6879(66)09034-7" @default.
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