Masuda M, Takamatsu S, Nishimura N, Komatsubara S, Tosa T
Research Laboratory of Applied Biochemistry, Tanabe Seiyaku Co., Ltd., Osaka, Japan.
Appl Biochem Biotechnol. 1992 Dec;37(3):255-65. doi: 10.1007/BF02788877.
Serratia marcescens T-2000 was previously reported to be an L-threonine-producing strain that harbors the recombinant plasmid carrying the mutant-type threonine operon. This strain produced 55 g of L-threonine/L of the medium containing urea as a nitrogen source after 72 h of cultivation. In the urea-containing medium, transitory stop of the growth was observed during the early period of cultivation when the entire amount of ammonium ion formed from urea via heat decomposition disappeared in the medium. This indicated that the shortage of ammonium supply in cells might delay both the cell growth and the L-threonine production. The use of ammonia water as a nitrogen source for L-threonine production was therefore studied, because microbial cells generally assimilate this source more readily than urea. When ammonia water was automatically fed to the medium so as to maintain the pH of the medium at around 7, the growth was accelerated, and the L-threonine production reached a maximum of 65 g/L at 48 h. Under these conditions, sucrose, a carbon source, was continuously fed to the medium, resulting in the production of 100 g of L-threonine/L at 96 h. Thus, the L-threonine production of the recombinant L-threonine-producing strain could be increased by devising the method for supply of a nitrogen source.
粘质沙雷氏菌T-2000先前被报道为一株产L-苏氨酸的菌株,它携带含有突变型苏氨酸操纵子的重组质粒。该菌株在以尿素作为氮源的培养基中培养72小时后,每升培养基可产生55克L-苏氨酸。在含尿素的培养基中,培养初期观察到生长出现短暂停滞,此时由尿素经热分解形成的铵离子在培养基中全部消失。这表明细胞中铵供应的短缺可能会延迟细胞生长和L-苏氨酸的产生。因此,研究了使用氨水作为L-苏氨酸生产的氮源,因为微生物细胞通常比尿素更容易同化这种氮源。当将氨水自动加入培养基以维持培养基pH值在7左右时,生长加速,L-苏氨酸产量在48小时时达到最高65克/升。在这些条件下,将碳源蔗糖持续加入培养基,96小时时L-苏氨酸产量达到每升100克。因此,通过设计氮源供应方法,可以提高重组产L-苏氨酸菌株的L-苏氨酸产量。