• Korean Journal of Food Science and Technology
• /
• v.4 no.2
• /
• pp.112-115
• /
• 1972

In the cource of investigation on L-glutamic acid production, acetate assimilating bacteria were isolated from natural sources. Among them, the strain No. 1214 was selected and characterized as a strain related to the genus Brevibacterium according to the standard method of taxonomy. This strain could grow in the acetate medium and accumulated a considerable amount of L-glutamic acid (22 g/L).

• Microbiology and Biotechnology Letters
• /
• v.16 no.2
• /
• pp.150-155
• /
• 1988

In order to produce L-lysine from cellulosic substrates by the intergeneric protoplast fusion between cellulolytic bacteria, Cellulomonas flavigena KFCC31221 and amino acid producing bacteria, Brevibacterium flavum ATCC14067, Corynebacteriurn glutamicum ATCC13032, conditions for protoplast formation and regeneration of these strains were investigated. After the strains were mutated with 500$\mu\textrm{g}$/$m\ell$ N-methyl-N'-nitro N-nitrosoguanidine for 30 min and the mutants were enriched by treating 300$\mu\textrm{g}$/$m\ell$ penicillin-G for 2 hrs, B. flavum Hse- Str$^{r}$ , C. glutamicum Met$^{-}$Thr$^{-}$ Rif$^{r}$ and Cellulomonas flavigena Thr$^{-}$Val$^{-}$Kan$^{r}$ were isolated. The rate of protoplast formation ranged from 95 to 98% when strains were treated at the concentration of 500$\mu\textrm{g}$/$m\ell$ of lysozyme, pH 6.5, 33$^{\circ}C$, for 6 hrs. in Tris- malate buffer supplemented with 0.4M sucrose as osmotic stabilizer. Approximately 30-33% protoplast was regenerated on the regeneration complete medium(RCM) containing 1.5% agar and 0.5M sodium succinate overlaid with the same medium except 0.7% agar.

• Microbiology and Biotechnology Letters
• /
• v.26 no.2
• /
• pp.110-116
• /
• 1998

The secY gene of Streptomyces lividans TK24 was cloned by the PCR method with synthetic oligonucleotide primers designed on the basis of the conserved regions of Ll5-secY-adk operon from E. coli, B. subtilis, and M luteus. The deduced amino acid sequences of the SecY are highly homologous to those of other known SecY. It has 46%, 43%, 57%, 44%, 42%,56%, 90% similarity to Escherichia coli, Bacillus subtilis, Micrococcus luteus, Bacillus licheniformis Staphylococcus carnosus, Brevibacterium flavum, Streptomyces scabies, respectively and almost the same with Streptomyces coelicolor, The gene organization of Ll5- SecY-Adk is also similar to those of other bacteria. SecY and Adk are very likely translationally coupled that is overlapping stop codon of SecY and start codon of Adk with one base pair, which is common structure among high GC content strains of gram positive bacteria.

• Microbiology and Biotechnology Letters
• /
• v.2 no.3
• /
• pp.141-147
• /
• 1974

The cultural conditions for L-glutamate production were investigated using Brevibacterium flavum nov. sp. D2209B, the most productive strain among 5 strains reported in preceeding paper. A temperature of 3$0^{\circ}C$ and a medium volume of 30 ml per 500-flask were selected as standard culture conditions. And the following results were obtained. 1. When the concentration of acetate in the medium was below 30 g per litre, the maximum amount of L-glutamate was accumulated. 2. KH$_2$PO$_4$, MgSO$_4$, FeCI$_3$ and MnCI$_2$ were required for the L-glutamate poduction, but the concentration of those inorganic salts little effected. 3. Signifcant amount of L-glutamate was accutnulated in the limited biotin concentration less than 0.3 ug per litre. 4. The addition of malic acid or succinic acid enhanced the accumulation. 5. The L-glutamate accumulation was related to the incubation time of seed; the amount of L-glutamate accumulated was maximum by inoculating 16-20 hour incubated seed. 6. In the medium containing sufficient amount of biotin for growth, L-glutamate accumulation was stimulated by the addition of penicillin at appropreate time during incubation.

• Microbiology and Biotechnology Letters
• /
• v.16 no.3
• /
• pp.175-181
• /
• 1988

To produce L-lysine from cellulosic substrate, the intergeneric protoplast fusion between Cellulomonas flavigena and Corynebacterium glutamicum, Cellulomonas flavigena and Brevibacterium flavum was performed. The fusion frequencies were 1.9$\times$10$^{-6}$ to 2.1$\times$10$^{-6}$ for the regenerated protoplasts when two parental strains were treated with 30% of polyethyleneglycol (M.W.6000) containing 5 mM EDTA at 3$0^{\circ}C$ for 30 min. Two fusants, FCB3 and FCC 19 were finally selected by comparision of their genetic stability and L-lysine productivity. The properties of fusants-DNA con-tent, G＋C content and L-lysine productivity-were investigated. The DNA content of fusants was greater than those of the parental strain and their G＋C contents are equal to half of total G＋C con-tent of two parental strains. The fusants showed high productivity of L-lysine from carboxy methyl cellulose as substrate.

• Microbiology and Biotechnology Letters
• /
• v.1 no.2
• /
• pp.105-113
• /
• 1973

The effect of biotin and biotin vitamer on the fermentative production of L-glutamic acid (L-GA) by Brevibacterium flavum was studied. And results were as follows. 1) L-GA production in the medium containing 10％ Glucose was the best at the concentration of Biotin 5${\gamma}$/l, Desthiobiotin 5${\gamma}$/1, and 7,8-Diaminopelargonic acid 10${\gamma}$/1, respectively. 2) In the experiment using the Glucose-Acetate mixed media derided into four parts, considerable amounts of cell growth and L-GA production were observed in the mixed medium containing 2％ Glucose-Acetate. 3) In the cases of using the media containing methanol, ethanol, ethylacetate, acetic acid (free acetate), Na-acetate:NH$_4$-acetate=2 : 1, the production of L-GA were in decreasing order as follows; Na-Acetate:NH-Acetate=2 : 1> Acetic acid (free acetate)> Ethylacetate> Ethanol＞ Methanol. 4) When biotin vitamers as growth factors were added in the medium containing Glucose or Acetate as the source of carbon, the substitution effect of Desthiobiotin was almost the same, 7,8-Diaminopelargonic acid 3 or 4 times stronger, and Bisnorbiotin has no substitution effect, compared with Biotin.

• Biotechnology and Bioprocess Engineering:BBE
• /
• v.7 no.3
• /
• pp.171-177
• /
• 2002

The metabolic flux pattern for L-histidine production was analyzed when glucose and/or acetate were used as carbon sources. Total L-histidine production was enhanced when mixed substrate (glucose and acetate) was used, compared wish that when either glucose or acetate was used as the sole carbon source. Theoretical maximum carbon fluxes through the main pathways for L-histldine production, cell growth, and ATP consumption for cell maintenance were obtained by the linear programming (LP) method. By comparison of the theoretical maximum carbon fluxes tilth actual ones, it was found that a large amount of glucose was actually used for maintenance of cell viability. On the other hand, acetate was used for cell growth. After depletion of acetate in the mixed substrate culture, the flux for glucose to L-histldine synthesis was markedly enhanced. A strategy for effective L-histidine production using both carbon sources was proposed.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.11 no.4
• /
• pp.213-217
• /
• 1978

Henri-Michaelis-Menten model for predicting the behavior of culture of Brevibacterium flavum under carbon limiting condition has been evaluated on a chemostat growing in the presence of the phenol derivatives: guaiacol, o-vanillin and vanillin. It is found that Henri-Michaelis-Menten model could be applicable to the evaluation of the growth rate of cells in the phenol derivatives. The marked enhancing abilities of the cells treated by the phenol derivatives during fermentation were ascribed to the formation of charge-transfer complex between the phenol-derivatives and oxygen which supplies oxygen effectively to the fermentation system.

• Microbiology and Biotechnology Letters
• /
• v.19 no.1
• /
• pp.31-36
• /
• 1991

A 7.5 kilobases hybrid plasmid, designated as pCE1301, was constructed by combining Eschurichia cwli plasmid pBELl which carries the kanamycin resistance gene of Tn5 with a cryptic plasmid, pSRl of Corynebacterium glutamicum. pCE1301 was transformed C. glutaicum by PEG-mediated protoplast method and its transformation efficiency was about $3.0\times 10^3$ transformants per $\mu g$ of the hybrid plasmid DNA. The physical map reveals that pCE1301 has single restriction sites for SalI and EcoRl, respectively. 'The kanamycin resistance of pCE1301 was stably maintained in C. glutamicum up to 25 generations and any segregation was not detected. pCI31301 was also introduced into Brevibacterium flavum and E coil, and replicated in those strains. pCE1301 was proved to be useiul in cloning the homoscrine dehydrogenase gene from C. glutamicum.

• Korean Journal of Food Science and Technology
• /
• v.21 no.1
• /
• pp.154-163
• /
• 1989

This studies were designed to improve the productivity of L-lysine by protoplast fusion and immobilized system of fusants using strains of Brevibacterium flavum ATCC 21528, Brevibacterium lactofermentum ATCC 21086 and Corynebacterium glutamicum 820. Mutants were isolated with concentration method of $300{\mu}g/ml$ penicillin-G after treatment of $250{\mu}g/ml$ N-methyl-N-nitro-N-nitrosoguanidine. B. flavum $37-2(Hos^-,\;Kan^r,\;AEC^r)$, B. lactofermentum $6-2(Ile^-,\;Val^-,\;Str^r,\;AEC^r)$ and C. glutamicum 57-5$(Met^-,\;Thr^-,\;Rif^r,\;AEC^r)$ were isolated from mutants. Protoplasts were induced by being incubated with $500{\mu}g/ml$ lysozyme of lysis solution for 6 hr and the ratio of protoplast formation and regeneration were ranging from 97-99% and 33-37%, respectively. Fusion frequencies of fusants of BBFL 21, BCFG 37 and BCLG 59 were shown in the range from $1.25{\times}10^{-6}\;to\;5.83{\times}10^{-7}$ under the optimum conditions. The fusant BBFL 21 showed the highest productivity of $411.1\;ng/ml{\cdot}hr$ L-lysine in the lysine productivity broth at $30^{\circ}C$ for 72hr. In the immobilization systems, fusant BBFL 21 was employed in various polymer matrices such as sodium alginate, polyacrylamide, agar and ${\alpha}-carrageena$. The immobilization of sodium alginate showed the highest productivity of $413\;ng/ml{\cdot}hr$ L-lysine in the batch system. Continuous fermentation of immobilization system by using tube fermentor was produced the highest productivity $416.7\;ng/ml{\cdot}hr $ L-lysine under optimum condition.