以葡萄糖为碳源,硫酸铵为氮源,15N邻氨基苯甲酸为前体物,利用酵母发酵合成15NL-色氨酸,是合成15NL-色氨酸的一种新方法。为了有效的利用含氮的化合物,实验过程中,采用15N标记的邻氨基苯甲酸为示踪剂,分析了N元素在15NL-色氨酸合成过程中的代谢通量。结果表明,93%外加的邻氨基苯甲酸,被转化为15NL-色氨酸。硫酸铵在促进菌体生长的同时,0.05%硫酸铵被用于合成邻氨基苯甲酸,并进一步被转化为L-色氨酸,5.95%硫酸铵被直接用于合成15NL-色氨酸。
Abstract
A new synthesis method for 15NL-tryptophan by yeast fermentation using glucose as carbon source, ammonium sulphate as nitrogen source, and 15N anthranilic acid as a precursor is described. In order to ensure efficient use of the nitrogen source compounds, the metabolic flux of elemental nitrogen was analyzed for L-tryptophan biosynthesis using the 15N anthranilic acid as a tracer in the experimental process. The results indicated that 93% of the L-tryptophan was derived from exogenous anthranilic acid. Ammonium sulphate contributed to the growth of the yeast and anthranilic acid was also synthesized from ammonium sulphate and subsequently converted into L-tryptophan (0.05% of the total yield). A further 5.95% of L-tryptophan was directly synthesized from ammonium sulphate.
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参考文献
[1] Ruhul Momen AZM, Tsutomu H. Biosynthesis of Violacein: Intact Incorporation of the Tryptophan Molecule on the Oxindole Side with Intramolecular Rearrangement of the Indole Ring on the 5-Hydroxyindole Side [J]. Biosci Biotechnol Biochem, 2000, 64(3): 539-549.
[2] rapparini F, Cohen JD, Slovin JP. Indole-3-acetic acid biosynthesis in Lemna gibba studied using stable isotope labeled anthranilate and tryptophan[J]. Plant Growth Regulation, 1999, 27: 139-144.
[3] Unkefer CJ, Lodwig SN, Silks LA et al. Stereoselective Synthesis of Stable Isotope-Labeled L-α-Amino Acids: Chemomicrobiological Synthesis of L-[β-13C]-, L-[2’-13C]-, and L-[1’-15N]Tryptophan [J]. Journal of Labelled Compounds and Radiopharmaceuticals, 1991,29: 1247-1256.
[4] 范如霖.稳定同位素标记有机合成法[M].北京,化学工业出版社,1986, 88-89.
[5] 王健,陈宁.基于途经分析及代谢流分析的L-色氨酸发酵条件优化[J].云南大学学报(自然科学版),2004,26(6A):68-73.
[6] 王健,陈宁,张蓓等.L-色氨酸生物合成的代谢流量的分析[J].微生物学报,2003,43(4):473-479.
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脚注
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