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在装有疏水化聚(丙烯酰胺)冷冻凝胶的柱中对烃氧化红球菌细胞的选择性吸附

Selective adsorption of hydrocarbon-oxidizing Rhodococcus cells in a column with hydrophobized poly(acrylamide) cryogel.

作者信息

Kuyukina Maria S, Rubtsova Ekaterina V, Ivshina Irena B, Ivanov Roman V, Lozinsky Vladimir I

机构信息

Institute of Ecology and Genetics of Microorganisms, Russian Academy of Sciences, Perm, Russia.

出版信息

J Microbiol Methods. 2009 Oct;79(1):76-81. doi: 10.1016/j.mimet.2009.08.003. Epub 2009 Aug 12.

Abstract

A method for selective adsorption of Rhodococcus cells in the column with hydrophobized poly(acrylamide) cryogel (cryoPAAG) was developed that allowed rhodococci separation from mixed bacterial populations and their effective concentration within a sponge-like gel matrix. Hydrophobization of cryoPAAG using the n-dodecane graft (C12) was performed to enhance the adhesion of Rhodococcus cells to the cryogel; this was suggested by our finding that alkanotrophic rhodococci possess high adhesive activity (91-98%) towards n-alkanes, whereas other Gram-positive and Gram-negative bacteria tested did not adhere strongly to hydrocarbons. The selective index of the hydrophobic C12-cryoPAAG column for Rhodococcus cells was 72% that ensured their separation from complex bacterial cultures. Respirometry results using the Columbus Micro-Oxymax respirometer showed that the maximal respiratory activity of C12-cryoPAAG-immobilized Rhodococcus cells incubated with petroleum hydrocarbons was 1.6-1.8 times higher than that of freely suspended cells, and this correlated with the largest immobilized cell number. Moreover, high respiration rates were maintained over 3 weeks of incubation, indicating a considerable functional stability of the cryoPAAG-immobilized biocatalyst developed.

摘要

开发了一种在柱中用疏水化聚(丙烯酰胺)冷冻凝胶(cryoPAAG)选择性吸附红球菌细胞的方法,该方法可使红球菌从混合细菌群体中分离出来,并在海绵状凝胶基质中有效浓缩。使用正十二烷接枝(C12)对cryoPAAG进行疏水化处理,以增强红球菌细胞与冷冻凝胶的粘附力;我们的研究发现表明了这一点,即烷营养型红球菌对正构烷烃具有较高的粘附活性(91-98%),而测试的其他革兰氏阳性和革兰氏阴性细菌对烃类的粘附力不强。疏水C12-cryoPAAG柱对红球菌细胞的选择指数为72%,确保了它们从复杂细菌培养物中的分离。使用哥伦布Micro-Oxymax呼吸仪的呼吸测定结果表明,与石油烃一起孵育的C12-cryoPAAG固定化红球菌细胞的最大呼吸活性比自由悬浮细胞高1.6-1.8倍,这与固定化细胞数量最多相关。此外,在孵育3周期间保持了高呼吸速率,表明所开发的cryoPAAG固定化生物催化剂具有相当大的功能稳定性。

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