Accumulation of 2-Keto-L-Gulonate at 33°C by a Thermotolerant Gluconobacter Oxydans Mutant Obtained by Ion Beam Implantation

Abstract

To obtain thermotolerant mutants of G. oxydans, which can enhance the transformation rate of L-sorbose to 2-Keto-L-gulonate (2-KLG) at 33oC in a two-step process of vitamin C manufacture, ion beam was used as a mutation source. Gluconobacter oxydans G0 and Bacillus megaterium B0 were used in this study. The original strain Gluconobacter oxydans G0 was mutated by the heavy ion implantation facility at the Institute of Plasma Physics, Chinese Academy of Sciences. Several mutants including Gluconobacter oxydans GI13 were isolated and cocultured with Bacillus megaterium B0 at 33oC in shaking flasks. The average transformation rate of the new mixed strain GI13-B0 in per gram-molecule reached 94.4% after seven passages in shaking flasks, which was increased by 7% when compared with the original mixed strain G0-B0 (Gluconobacter oxydans G0 and Bacillus megaterium B0). Moreover, the transformation rate of I13B0 was stable at 94% at temperatures ranging from 25oC to 33oC, which would be of much value in reducing energy consumption in the manufacture of L-ascorbic acid, especially in the season of summer. To clarify some mechanism of the mutation, the specific activities of L-sorbose dehydrogenase in both G0 and GI13 were estimated.