Genetic Analysis of Recombinants by Interspecific Protoplast Fusion of Coryneform Bacteria and Their L-glutamate & L-glutamine Production

Corynebacterium 세균의 이종간 원형질체 융합에 의한 재조합주의 유전학적 분석과 L-glutamate와 L-glutamine 생성

For interspecific portoplast fusion, Brevibacterium flauum lOAHR (Rifr axg his) and Corynebacterium glutamicum 11TS ($Sm-r$ trp) were induced by UV and NTG treatment. The protoplast fusion frequency between E. flavum XOAHR and C. glutamicum llTS was $3.7\times 10^{-6}$ with the lysozyme treatment (300 P $\mu g$ml) for 18 hrs. Genotypes of recombinants were analized as FMM ($Rif^r\; Sm^r$), FA (Rift $Sm^r$ arg), FH ($Rif^r\; Sm^r$ his), FT ($Rif^r\; Sm^r$ trp), FAH ($Rif^r\; Sm^r$ arg trp), FAT ($Rif^r\; Sm^r$ arg trp), and FAHT ($Rif^r\; Sm^r$ arg his trp). FAH 1 produced 12 fold of glutamate production compared to parental type, E. flauum 10AHR. In glutamine productivity, it produced 2.6 fold to parental type, C. glutamicum 11TS. Production of glutamate or glutamine by recombinants was involved in the specific activities of glutamate dehydrogenase (GDH) and glutamine synthetase (GS), respectively.

자외선 조사와 NTG를 처리하여 Brevibacterium flavum 10AHR(arg his $Rif^r$)과 Corynebacterium glutamicum 11TS(trp $Sm^r$의 돌연변이주를 분리하였다. B.flavum 10AHR과 C.glutamicum 11TS를 300$\mu g$/ml의 lysozyme으로 18시간 처리하여 원형질체를 형성하고, 융합시 30의 PEG 6,000으로 처리하였을 때 가장 높은$3.7\times 10^{-6}$의 융합빈도를 나타내었다.