反相高效液相色谱法分析盘基网柄菌对氨基酸的利用  

Analysis of Amino Acid Consumption During the Cultivation of Dictyostelium discoideum by Reversed-phase High Performance Liquid Chromatography

作　　者：黄霄红[1] 王颖[2] 叶美玲[2] 罗臻[2] 宋娟[3] 凌雪萍[2] 

机构地区：[1]厦门大学医学院,福建厦门361005 [2]厦门大学化学化工学院,福建厦门361005 [3]吉林华康药业股份有限公司,吉林敦化133700

出　　处：《厦门大学学报（自然科学版）》2012年第4期734-739,共6页Journal of Xiamen University：Natural Science

基　　金：国家自然科学基金项目(31070296)

摘　　要：为观察盘基网柄菌(Dictyostelium discoideum)在SIH培养基上对氨基酸的利用情况,以2,4-二硝基氯苯为衍生化试剂,研究了用反相高效液相色谱(RP-HPLC)测定氨基酸的方法.在60min内16种氨基酸达到基线分离,峰面积与氨基酸浓度的线性相关系数为0.992～0.999.对发酵液样品的分析结果表明,盘基网柄菌对赖氨酸的利用最为彻底,发酵后期赖氨酸已被消耗完全.蛋氨酸、色氨酸、精氨酸、组氨酸也较易被盘基网柄菌利用,而对天冬氨酸、谷氨酸、甘氨酸、苏氨酸、脯氨酸、缬氨酸的需求不大.这一代谢特点为改进盘基网柄菌培养基组成提供依据.In order to observe amino acid consumption during cultivation of Dictyosteliurn discoideurn on synthetic SIH medium, a method for determination of amino acids by reversed-phase high performance liquid ehromatography（RP-HPLC） with 2,4-dinitro- chlorobenzene derivatization was investigated. 16 amino acids could be well separated in 60 min, and a good linear relationship between peak area and concentration of amino acids was established with the correlation coefficients in the range of 0. 992-0. 999. Analysis re suits of samples of D. discoideum on SIH medium indicated that lysine was completely consumed after 7 d of cultivation. Methionine, tryptophan,arginine and histidine were also utilized considerably while aspartie acid, glutamic acid, glycine, threonine, proline and valine did not show significant demand during the cell growth. This metabolic characteristic would supply reliable data to design a more reasonable synthetic medium for D. discoideum.

关 键 词：盘基网柄菌 反相高效液相色谱法 氨基酸 2 4-二硝基氯苯 

分 类 号：Q815[生物学—生物工程]