_L-Alanine Fermentation by an Alanine Racemase-Deficient Mutant of the _<DL>-Alanine Hyperproducing Bacterium Arthrobacter oxydans HAP-1
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概要
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Arthrobacter oxydans HAP-1 produces a large amount of _<DL>-alanine from glucose via the reductive amination of pyruvate catalyzed by _L-alanine dehydrogenase(ALD). This bacterium was found to be capable of growing on either _D- or _L-alanine as a sole carbon source with lower growth rates than on glucose. The effects of an alanine racemase(AR)inhibitor on the accumulation and/or consumption of _D- or _L-alanine by the bacterium suggested that _D-alanine was formed by the isomerization of _L-alanine and degraded through the reverse reaction. _D-Alanine non-utilizing mutants were derived from strain HAP-1 by the staining method using the chromogenic agent 2, 3, 5-triphenyl tetrazolium chloride. The mutants were obtained were of two kind: ALD-deficient and AR-deficient. The former lost the ability to grow on both _D- and _L-alanine, indicating that the ALD of strain HAP-1 functions not only in _L-alanine synthesis but also in _L-alanine degradation. The latter could utilize _L-alanine as a carbon source but not _D-alanine; furthermore, they required _D-alanine for growth in minimum or nitrient medium. A representative AR-deficient mutant, DAN 75, produced 75.6 g of _L-alanine per liter with high optical purity(97% e.e.)in a fed-batch cultivation(glucose was fed to 15% in total)using a jar-fermentor. The _L-alanine titer was almost equal to the sum of _D- and _L-alanine produced by the parental strain. The production rate of DAN 75, as well as that of the parental strain HAP-1, slowed down in the late stage of the cultivation. Examination of the influence of exogenously added alanine revealed this reduced production to be due to a deceleration in the glucose consumption rate arising from alanine accumulated in the medium.
- 1998-10-25
著者
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Hashimoto Shin-ichi
Tokyo Research Laboratories Kyowa Hakko Kogyo Co.ltd.
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KATSUMATA RYOICHI
Tokyo Research Laboratories, Kyowa Hakko Kogyo Co.Ltd.
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Katsumata Ryoichi
Tokyo Research Laboratories Kyowa Hakko Kogyo Co.ltd.:(present Address)laboratory Of Animal Microbio
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Katsumata Ryoichi
Tokyo Researach Laboratories Kyowa Hakko Kogyo Co. Ltd.
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- _L-Alanine Fermentation by an Alanine Racemase-Deficient Mutant of the _-Alanine Hyperproducing Bacterium Arthrobacter oxydans HAP-1
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