Abstract
End-product inhibition is a key factor limiting the production of organic acid during fermentation. Two rounds of heavy-ion beam irradiation may be an inexpensive, indispensable and reliable approach to increase the production of butyric acid during industrial fermentation processes. However, studies of the application of heavy ion radiation for butyric acid fermentation engineering are lacking. In this study, a second 12C6+ heavy-ion irradiation-response curve is used to describe the effect of exposure to a given dose of heavy ions on mutant strains of Clostridium tyrobutyricum. Versatile statistical elements are introduced to characterize the mechanism and factors contributing to improved butyric acid production and enhanced acid tolerance in adapted mutant strains harvested from the fermentations. We characterized the physiological properties of the strains over a large pH value gradient, which revealed that the mutant strains obtained after a second round of radiation exposure were most resistant to harsh external pH values and were better able to tolerate external pH values between 4.5 and 5.0. A customized second round of heavy-ion beam irradiation may be invaluable in process engineering.
Highlights
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Improved strains have been generated by heavy-ion beam irradiation and selection for the development of commercial strains for use in microbial fermentation processes16–19. 12C6+ heavy-ion irradiation may represent an inexpensive and reliable approach for increasing the productivity of industrial processes
We observed that carbon ions effectively induced the expression of key enzymes in glycolysis, product formation, energy and redox metabolism, and the cellular response to butyric acid production
Summary
Other related technologies has provided new tools for approaching yields improvement by means of genetic manipulation of biosynthetic pathway. It is crucial to distinguish mutants that are acid-tolerant, and well-adapted mutant strains were characterized for their physiological properties, including their ability to survive 12C6+ heavy-ion irradiation, expression status during the fermentation process, cell growth, changes in response to pH variation, butyric acid tolerance, butyrate/acetate ratio, and gas production. Evaluating these characteristics often involves a quantitative approach to the acquisition and analysis of data, and an efficient discovery process is required to extract information from the data. The introduction of versatile statistical methods enabled a thorough characterization of the productivity and acid tolerance of the secondary mutant
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