生物技术
生物技術
생물기술
BIOTECHNOLOGY
2010年
1期
27-30
,共4页
李云%杨胜远%杨韵晴%黄荣城%陈郁娜%刘祥流
李雲%楊勝遠%楊韻晴%黃榮城%陳鬱娜%劉祥流
리운%양성원%양운청%황영성%진욱나%류상류
γ-氨基丁酸%谷氨酸脱羧酶%屎肠球菌%鉴定
γ-氨基丁痠%穀氨痠脫羧酶%屎腸毬菌%鑒定
γ-안기정산%곡안산탈최매%시장구균%감정
glutamate decarboxylase%γ- aminobutyric acid%Enterococcus faecium%identification
目的: 鉴定1株产γ氨基丁酸(γ-aminobutyric acid,GABA)的乳酸菌HS3,并研究了其谷氨酸脱羧酶(Glutamate decarboxylase,GAD)粗酶酶学性质.方法:根据形态培养特征、生理生化特征和16S rDNA序列比对及系统发育分析对菌株HS3进行了鉴定.采用菌体细胞破碎后的粗酶液,研究了温度、pH和金属离子对酶活的影响.结果:菌株HS3的形态培养和生理生化特征符合肠球菌属(Enterococcus)特征,其16S rDNA序列与Enterococcus faecium(EU717962)16S rDNA序列同源性达99%,鉴定菌株HS3为屎肠球菌(Enterococcus faecium),菌株HS3 GAD最适作用温度为40 ℃,最适作用pH4.5.酶的热稳定较好,50℃处理4h,在pH3.5～6.0酶活基本稳定.Ca~(2+)对酶有激活作用,5mmol/L和50mmol/L浓度酶活分别提高了37.41%和17.43%.Ba~(2+)和Zn~(2+)在5mmol/L浓度时激活作用明显,而Mg~(2+)在5mmol/L浓度激活作用较好.结论:菌株HS3的GAD活力较高,稳定性较好,为生物合成GABA提供了新的微生物菌种资源.
目的: 鑒定1株產γ氨基丁痠(γ-aminobutyric acid,GABA)的乳痠菌HS3,併研究瞭其穀氨痠脫羧酶(Glutamate decarboxylase,GAD)粗酶酶學性質.方法:根據形態培養特徵、生理生化特徵和16S rDNA序列比對及繫統髮育分析對菌株HS3進行瞭鑒定.採用菌體細胞破碎後的粗酶液,研究瞭溫度、pH和金屬離子對酶活的影響.結果:菌株HS3的形態培養和生理生化特徵符閤腸毬菌屬(Enterococcus)特徵,其16S rDNA序列與Enterococcus faecium(EU717962)16S rDNA序列同源性達99%,鑒定菌株HS3為屎腸毬菌(Enterococcus faecium),菌株HS3 GAD最適作用溫度為40 ℃,最適作用pH4.5.酶的熱穩定較好,50℃處理4h,在pH3.5～6.0酶活基本穩定.Ca~(2+)對酶有激活作用,5mmol/L和50mmol/L濃度酶活分彆提高瞭37.41%和17.43%.Ba~(2+)和Zn~(2+)在5mmol/L濃度時激活作用明顯,而Mg~(2+)在5mmol/L濃度激活作用較好.結論:菌株HS3的GAD活力較高,穩定性較好,為生物閤成GABA提供瞭新的微生物菌種資源.
목적: 감정1주산γ안기정산(γ-aminobutyric acid,GABA)적유산균HS3,병연구료기곡안산탈최매(Glutamate decarboxylase,GAD)조매매학성질.방법:근거형태배양특정、생리생화특정화16S rDNA서렬비대급계통발육분석대균주HS3진행료감정.채용균체세포파쇄후적조매액,연구료온도、pH화금속리자대매활적영향.결과:균주HS3적형태배양화생리생화특정부합장구균속(Enterococcus)특정,기16S rDNA서렬여Enterococcus faecium(EU717962)16S rDNA서렬동원성체99%,감정균주HS3위시장구균(Enterococcus faecium),균주HS3 GAD최괄작용온도위40 ℃,최괄작용pH4.5.매적열은정교호,50℃처리4h,재pH3.5～6.0매활기본은정.Ca~(2+)대매유격활작용,5mmol/L화50mmol/L농도매활분별제고료37.41%화17.43%.Ba~(2+)화Zn~(2+)재5mmol/L농도시격활작용명현,이Mg~(2+)재5mmol/L농도격활작용교호.결론:균주HS3적GAD활력교고,은정성교호,위생물합성GABA제공료신적미생물균충자원.
Objective:A lactic acid bacterium (LAB) for γ - amino butyric acid (CABA) production was identified. The characterization of its glutamate decarboxylase ( GAD) was also reported in this paper. Method: The strain HS3 was identified base on morphological,physiological and biochemical characteristics, 16S rDNA and phylogenic analysis. The characterization of its GAD was investigated by using cell disruption fluid as crude GAD. Result:The morphological, physiological and biochemical characteristics of strain HS3 accorded with the characteristics of Enterococcus. The similarity of the 16S rDNA sequences between strain HS3 and Enterococcus faecium (EU717962) was up to 99%. The strain HS3 was identified as Enterococcus faecium. The optimal temperature and pH value for the GAD activity was 40 ℃ and pH 4.5 respectively. The GAD from strain HS3 was stable under 50℃ for 4h and at pH 4.0-6.5. 5mmol/L and 50mmol/L of Ca~(2+) significantly increased GAD activity, which promoted 37.41% and 7.43% of the GAD activity respectively. The activation were also observed on the concentration of 5mmol/L of Ba~(2+) , 5mmol/L of Zn~(2+) and 50mmol/L of Mg~(2+) , respectively. Conclusion:The high activity and stable GAD from strain HS3 accounts for the new strain resources of GABA biosynthesis by LAB.
