Kokufuta E, Matsumoto W, Nakamura I
Institute of Applied Biochemistry, The University of Tsukuba, Sakura-mure, Niihari-gun, Ibaraki 305, Japan.
Biotechnol Bioeng. 1982 Jul;24(7):1591-603. doi: 10.1002/bit.260240712.
Cell immobilization with polyelectrolyte complex prepared from strongly polyacidic and polybasic ions was investigated for cells from Nitrosomonas europaea (ATCC 25978). Trimethylammonium glycol chitosan (TGCI) and potassium poly(vinyl-alcohol) sulfate (KPVS) were used. The immobilization was carried out by directly mixing both polymer solutions with the culture broth: An excess of TGCI was first added to the culture broth to aggregate the cells, and then KPVS was added to form the complex with the excess TGCI and to entrap the aggregates with the resulting complex. From physiocochemical studies on the cell aggregation, the mechanism can be interpreted in terms of the adsorption of polyion caused by the salt linkages of the ionizable groups on the cell surface. The result of an electron microscopic observation showed that the cells are situated in the pores and on the surface of the complex support. When the immobilized cells were incubated in a medium buffered by phosphate and containing ammonium sulfate, a considerable amount of nitrite was formed; this was shown to be caused by the entrapped cells and also those cells released from the support and grown in the medium. The ammonia-oxidizing activity was retained even after a total of 200 h of incubation in a batch reactor. No deformation of the complex support was observed.
研究了用由强聚酸性和聚碱性离子制备的聚电解质复合物固定化欧洲亚硝化单胞菌(ATCC 25978)细胞的方法。使用了三甲基铵基壳聚糖二醇(TGCI)和聚乙烯醇硫酸钾(KPVS)。通过将两种聚合物溶液与培养液直接混合来进行固定化:首先向培养液中加入过量的TGCI以使细胞聚集,然后加入KPVS与过量的TGCI形成复合物,并将聚集体包埋在所得复合物中。从对细胞聚集的物理化学研究来看,该机制可以用细胞表面可电离基团的盐键引起的聚离子吸附来解释。电子显微镜观察结果表明,细胞位于复合物载体的孔内和表面。当将固定化细胞在由磷酸盐缓冲并含有硫酸铵的培养基中培养时,形成了大量亚硝酸盐;这表明是由包埋的细胞以及从载体释放并在培养基中生长的细胞引起的。即使在间歇反应器中总共培养200小时后,氨氧化活性仍得以保留。未观察到复合物载体变形。