CAJ | 학술논문

从分别生长于含Mn和Cr培养基中的棕色固氮菌(Azotobacter vinelandii Lipmann)突变种UW3分离纯化出MnFo和CrFe蛋白.为适应包括固氮酶在内的氧敏感蛋白的空间晶体生长的要求,应用简易而适用的厌氧加样装置代替固氮酶实验室所用的笨重厌氧箱(dry box),在地面进行厌氧加样.在充满氮气的简便有机玻璃箱内厌氧加样的所有样品中,分别用液/液扩散法和汽相扩散的坐滴法都可在一周内使MnFe和CrFe蛋白在宇宙飞船上从溶液中结晶出来.在所用的数种蛋白沉淀剂中,飞船上形成的所有晶体都为单晶,而地面上在多数沉淀剂中部生成大量孪晶.在相同沉淀剂中用液/液扩散法,飞船上生成CrFe蛋白的最大晶体比地面生成的最大晶体大1倍.而在相同沉淀剂中用汽相扩散的坐滴法,飞船上生成的MnFe蛋白最大晶体却没有地面生成的最大晶体大.这种差异也许是由不同结晶方法而不是不同蛋白所引起的.
종분별생장우함Mn화Cr배양기중적종색고담균(Azotobacter vinelandii Lipmann)돌변충UW3분리순화출MnFo화CrFe단백.위괄응포괄고담매재내적양민감단백적공간정체생장적요구,응용간역이괄용적염양가양장치대체고담매실험실소용적분중염양상(dry box),재지면진행염양가양.재충만담기적간편유궤파리상내염양가양적소유양품중,분별용액/액확산법화기상확산적좌적법도가재일주내사MnFe화CrFe단백재우주비선상종용액중결정출래.재소용적수충단백침정제중,비선상형성적소유정체도위단정,이지면상재다수침정제중부생성대량련정.재상동침정제중용액/액확산법,비선상생성CrFe단백적최대정체비지면생성적최대정체대1배.이재상동침정제중용기상확산적좌적법,비선상생성적MnFe단백최대정체각몰유지면생성적최대정체대.저충차이야허시유불동결정방법이불시불동단백소인기적.
Nitrogenase CrFe protein and MnFe protein were purified from a mutant strain UW3 ofAzotobacter vine/andii Lipmann grown on a medium containing Cr and Mn, respectively. In order to meetthe requirement for crystal growth of O2-susceptible proteins including nitrogenase in space, crystalliza-tion conditions were optimized for the proteins using a simple and suitable device, as a replacement for thecumbersome anaerobic box (dry box), for anaerobic addition of the protein samples. In all used precipitantand protein solutions added in the simplified plexi glass box, CrFe protein and MnFe protein could becrystallized on the spacecraft in one week by the liquid/liquid diffusion method and vapor diffusion by thesitting drop method, respectively. All formed crystals were single on the spacecraft, but under the samecondition twin crystals appeared on the ground. The size of the largest crystal grown in space from CrFeprotein was 2-fold larger than that on the ground. But the size of the largest crystal grown in space from MnFe protein was not larger than that on the ground. The difference in crystal growth in space betweenCrFe protein and MnFe protein could be resulted from the crystallization method, rather than the kind ofprotein.