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为检测产聚 β-羟基丁酸酯 (PHB)的重组大肠杆菌VG1(p TU14)中表达的透明颤菌血红蛋白 (VHb) ,建立了改进的 CO差光谱法。结果表明 ,PHB对 VHb的检测有较严重的干扰。 VG1(p TU14)的全细胞悬浊液经过 4次冻融、低速离心和过量的 Na2 S2 O4还原后 ,在试样端通入 CO进行3m in络合反应并在黑暗条件下静置 5 min,最后进行 VHb的 CO差光谱分析。当菌浓小于 2 5 m g/ m L缓冲液时 ,其定量关系符合 L am ber- Beer定律 ,可以准确地定量分析 VHb
To detect Vitreoscilla hemoglobin (VHb) expressed in recombinant β-hydroxybutyrate (PHB) -producing recombinant E. coli VG1 (pTU14), improved CO differential spectroscopy was established. The results showed that PHB had a more serious interference with the detection of VHb. The whole cell suspension of VG1 (pTU14) was subjected to four freeze-thaw cycles, low-speed centrifugation and excess Na2 S2 O4 reduction. CO was introduced into the sample for 3 minutes and allowed to stand in the dark for 5 minutes , The last VHb CO difference spectrum analysis. When the bacteria concentration is less than 25mg / ml buffer, its quantitative relationship with LAM ber-Beer law, can be accurately quantitative analysis of VHb