ABSTRACT The purpose of this research was to enhance the production of vitamin K2 by fermentation optimization and Arachis hypogaea supplementation in Flavobacterium sp. mutant SP-L-01. Optimized culture condition were as follows: 6-days shake-flask culture at 37oC with initial pH value 7.0 ± 0.2, shaking speed in 120 r/min and medium volume of 30 mL with 2% inoculums. After optimization of fermentation medium by response surface methodology (RSM), optimized medium were maltose 23.8 g/l, glucose 9.69 g/l, beef extract 15 g/l, K2HPO4 4.5 g/l，NaCl 3.0 g/l and MgSO4·7H2O 0.3 g/l. Production of vitamin K2 after optimization reached to 10.97 mg/l, which is 79.25% higher than that before optimization (6.12 mg/l). 3 mg/mL of arachis hypogaea was added into the medium at 72 h of shake-flake cultivation, which improved the production of menaquinone-4 (MK4) up to 371% and menaquinone-6 (MK6) up to 149% higher than those of the original medium. D-(+)-catechin, one of the components of arachis hypogaea, was added alone into the medium, which also improved the vitamin K2 synthesis.

The aim of this study was to enhance the production of vitamin K2 by using N-methyl-N-nitro-N-nitroso-guanidine (NTG) and low energy ion beam implantation and optimizing the fermentation medium. Mutation resulted in 1.66-fold higher production of vitamin K2 than that of the parentl strain. The production by the mutant BN-P15-11-1was increased 55% and reached 3.593±0.107 mg/L by using the Plackett-Burman and Box-Behnken designs to optimize the fermentation medium. The optimal fermentation culture medium was composed of (g/L) glycerol 69.6, sucrose 34.5, K2HPO4 4.0, peptone 20, yeast extract 25 and fermented at 37 °C and 150 rpm for 72 h. The results showed that the NTG and low energy ion beam implantation mutations and optimizing fermentation medium were effective methods to enhance vitamin K2production.

This work aimed to improve tannase yield of Aspergillus niger through N+ ion beam implantation in submerged fermentation. The energy and dose of N+ ion beam implantation were investigated. The results indicated that an excellent mutant was obtained through nine successive implantations under the conditions of 10 keV and 30-40 (×2.6×10(13)) ions/cm², and its tannase yield reached 38.5 U/mL, which was about five-time higher than the original strain. The study on the genetic stability of the mutant showed that its promising performance in tannase production could be stable. The studies of metal ions and surfactants affecting tannase yield indicated that manganese ions, stannum ions, xylene and SDS contained in the culture medium had positive effects on tannase production under submerged fermentation. Magnesium ions, in particular, could enhance the tannase yield by the mutant increasing by 42%, i.e. 53.6 U/mL. Accordingly, low-energy ion implantation could be a desirable approach to improve the fungal tannase yield for its commercial application.