生物技术
生物技術
생물기술
BIOTECHNOLOGY
2009年
6期
10-14
,共5页
脱枝酶%分离%鉴定%酶学性质
脫枝酶%分離%鑒定%酶學性質
탈지매%분리%감정%매학성질
debranching enzyme%screening%identification%enzymatic properties
目的:从中国高校工业微生物资源与信息中心(CICIM-CU)细菌库中分离具有产脱枝酶酶活的细菌并鉴定,进行酶学性质的研究.方法:通过碘显色平板法筛选产酶菌株,利用16S rDNA确定其属种.对每一株产脱枝酶的细菌进行初步的酶学性质研究.结果:从4005株细菌中筛选出45株产脱枝酶的细菌,建立了产脱枝酶细菌的细菌库.酶学性质表明CICIM B272、CICIMB1-30两株菌产生的脱枝酶,酶反应的最适温度分别为70℃、60℃,最适pH分别为6.0、5.5,来源于上述两种不同属种的脱枝酶在30-70℃反应条件下,酶在pH4.5～8.5范围内活性稳定,Li~+、Na~+、K~+、M~(2+)、Mn~(2+)对两者酶活均有显著的激活作用,而Cu~(2+)、Fe~(3+)及EDTA对两者均有显著的抑制作用,Mn~(2+)、Ca(2+)分别对两者的热稳定性具有很好的提升作用.以支链淀粉为底物的动力学常数K_m分别为352.883mg/mL、4.5814mg/mL,V_(max)分别为30.03mg/min·mL、0.4575mg/min·mL.结论:不同属种的脱枝酶酶学性质差别显著.
目的:從中國高校工業微生物資源與信息中心(CICIM-CU)細菌庫中分離具有產脫枝酶酶活的細菌併鑒定,進行酶學性質的研究.方法:通過碘顯色平闆法篩選產酶菌株,利用16S rDNA確定其屬種.對每一株產脫枝酶的細菌進行初步的酶學性質研究.結果:從4005株細菌中篩選齣45株產脫枝酶的細菌,建立瞭產脫枝酶細菌的細菌庫.酶學性質錶明CICIM B272、CICIMB1-30兩株菌產生的脫枝酶,酶反應的最適溫度分彆為70℃、60℃,最適pH分彆為6.0、5.5,來源于上述兩種不同屬種的脫枝酶在30-70℃反應條件下,酶在pH4.5～8.5範圍內活性穩定,Li~+、Na~+、K~+、M~(2+)、Mn~(2+)對兩者酶活均有顯著的激活作用,而Cu~(2+)、Fe~(3+)及EDTA對兩者均有顯著的抑製作用,Mn~(2+)、Ca(2+)分彆對兩者的熱穩定性具有很好的提升作用.以支鏈澱粉為底物的動力學常數K_m分彆為352.883mg/mL、4.5814mg/mL,V_(max)分彆為30.03mg/min·mL、0.4575mg/min·mL.結論:不同屬種的脫枝酶酶學性質差彆顯著.
목적:종중국고교공업미생물자원여신식중심(CICIM-CU)세균고중분리구유산탈지매매활적세균병감정,진행매학성질적연구.방법:통과전현색평판법사선산매균주,이용16S rDNA학정기속충.대매일주산탈지매적세균진행초보적매학성질연구.결과:종4005주세균중사선출45주산탈지매적세균,건립료산탈지매세균적세균고.매학성질표명CICIM B272、CICIMB1-30량주균산생적탈지매,매반응적최괄온도분별위70℃、60℃,최괄pH분별위6.0、5.5,래원우상술량충불동속충적탈지매재30-70℃반응조건하,매재pH4.5～8.5범위내활성은정,Li~+、Na~+、K~+、M~(2+)、Mn~(2+)대량자매활균유현저적격활작용,이Cu~(2+)、Fe~(3+)급EDTA대량자균유현저적억제작용,Mn~(2+)、Ca(2+)분별대량자적열은정성구유흔호적제승작용.이지련정분위저물적동역학상수K_m분별위352.883mg/mL、4.5814mg/mL,V_(max)분별위30.03mg/min·mL、0.4575mg/min·mL.결론:불동속충적탈지매매학성질차별현저.
Objective: The bacteria of producing debranching enzyme were screened from CICIM-CU, identified, and studied the characteristics of debranching enzyme. Method:Screened debranching enzyme producing strain, identified taxonomic status, determine debranching enzyme properties by iodine solution coloration plate and 16S rDNA sequence alignment method. Result:45 strains which can produce debranching enzyme were screened from 4005 strains and the bacteria bank capable to produce debranching enzyme was set up finally.The results showed that the debranching enzyme from CICIM B272 and CICIM B1-30 performed the maximal activity at 70℃, pH 6.0 and 60℃ pH 5.5, respectively. The enzyme had thermostability between 30℃ and 70℃ and pH stability from 4.5 to 8.5. The enzymes were strongly activated by Li~+, Na~+, K~+, Mg~(2+), Mn~(2+), whereas inhibited by Cu~(2+), Fe~(3+) and EDTA. Mn~(2+), Ca~(2+) significantly increased their thermostability, respectively. The kinetic constants of the enzyme was obtained, their K_m for amylopectin were 352.883mg/mL and 4.5814mg/mL, V_(max) were 30.03 mg/min·mL and 0.4575 mg/min·mL, respectively. Conclusion: The difference in the enzymatic property of debranching enzyme was significant between different genera.
