RESUMEN
Abstract A new plate method was developed for rapid screening of Ketogulonicigenium vulgare mutants overproducing 2-keto-l-gulonic acid (2-KLG). The screening methodology took the advantage of the acidity caused by 2-KLG, which changes the color of bromothymol blue (pH indicator) from blue to yellow. Using the proposed method, a mutant, K. vulgare 65, was selected from 20,000 colonies produced by a strain subjected to spaceflight mutagenesis. When co-cultured with Bacillus megaterium 2980 in 20-L fermenters, K. vulgare 65 showed a high conversion rate (94.45%) of l-sorbose to 2-KLG. In contrast to the traditional screening method, this one significantly improved the frequency of obtaining positive mutants. The proposed plate screening method is cost-effective and easy to run and is thus useful for the isolation and screening of K. vulgare mutants overproducing 2-KLG.
Asunto(s)
Azúcares Ácidos/metabolismo , Técnicas Bacteriológicas/métodos , Rhodobacteraceae/metabolismo , Sorbosa/metabolismo , Técnicas Bacteriológicas/instrumentación , Rhodobacteraceae/aislamiento & purificación , Rhodobacteraceae/genética , Fermentación , MutaciónRESUMEN
Objective To study mutagenic effects of spaceflight on physiological and biochemical parameters of Rhodobacter sphaeroides and select high-yield mutants in co-enzyme Q10(CoQ10) production for providing the experimental and theoretical basis for industrial production through mutagenic effects of spaceflight.Methods Variations in stress resistance and CoQ10 production of isolated strains were studied,the strain Rhodobacter sphaeroides was taken back by a recoverable satellite after 15 d flight in space.Results Compared to the control,the strain was characterized by highter NaCl tolerance and higher stress resistance,as well as with broader scope in growth temperature and pH value after spaceflight.The mutant colonies appeared white or pink which was different from their original red.The CoQ10 production of mutant 10 strain was increased by 73.13% much higher than that of control.Conclusion Spaceflight mutagenic effects on Rhodobacter sphaeroides shows to be multi-factor compared to the traditional single-factor mutagenesis methods.It can enhance stress resistance and increase CoQ10 production of isolated strains,and can be utilized in industrial microbial mutagenesis and breeding in the future.